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2. Measure rectangle 8 HL Turn on off Parameter Control Bar 1 Adjust calibration magnification Turn on off Sequencer Control Bar Set target temperature Turn on off Data Control Bar Set target inflow pressure Turn on off Analysis Control Bar Set target outflow pressure Ble fe Stop start image display Reset analysis Save image to disc Set analysis rectangle E E R E R E BA Configure Frame Grabber Stop start analysis Zoom in Hi Configure analysis Full size About Calibrate image H Start VEDIVIEW Helpdesk nil A ey Figure 4 9 The Meaning and action of each icon User Manual vers 3 2 Chapter 4 39 4 2 4 Main Window The main window can show any of the graphical displays of the control bars which are frame grabber data or analysis The main window display may be configured from the view menu or simply by a mouse click on the relevant control bar display The rel evant popup menu for each control bar can be revealed by a right mouse button click when the Windows cursor is anywhere in the main window The three options for the main window are shown below Figure 4 10 Main window displaying the framegrabber images overlaid with the vessel dimentions found by activation of the vessel detedtion and analysis Figure 4 11 Main window dis playing the data history Figure 4 12 Main window dis playing the quantitative result of image analysis in fo
3. the Pressure Regulator the Microscope and the PC are switched off 1 Myograph Unit Myo Interface Connection Connect the Pressure Myograph Unit to the Myo Interface by using the grey 25 pin connection cable The end of the cable with the temperature probe is used with the Pressure Myograph Unit 2 Myo Interface PC Connection Connect the Myo Interface to the computer with the 9 pol serial cable from the COM port RS 232 on the back panel of the Myo Interface to the COM port RS 232 on the computer 3 Myo Interface Pressure Regulator Connection Connect the Myo Interface to the Pressure Regulator by using the grey 15 pin connection cable Connect the power cord to the power inlet on the back panel of both the Myo In terface and the Pressure Regulator 4 Pressure Regulator Oxygen Supply Connection Connect the Main Pressure Inlet on the back panel of the Pressure Regulator to an adjustable oxygen supply using the Atlas Copco Cablair hose delivered with the Pressure Myograph System Important Make sure that the hose is securely fastened to the Main Pressure Inlet using the hose clip The pressure from the oxygen supply must not exceed 1 0 Bar User Manual vers 3 2 Chapter 2 19 20 5 Microscope Camera PC Connection Model 111P Connect the DSUB 9 pol cable from the DMT Microscope to the 9 pol port on the frame grabber card installed in the computer Model 110P Connect the D SUB 15
4. dissolve the EDTA 2 Solution 1 is added to a graduated bottle and the bottle is filled with double distilled H O to a final volume of 500ml 3 Solution 3 is added to the graduated bottle which afterwards is filled with additional double distilled HO to a final volume of about 850ml 4 Aerate the solution with carbogen 95 0 5 CO for about 20 minutes 5 Solution 2 is added and the graduated bottle is filled with additional double distilled H O to reach the final volume of 1000ml Continue the carbogen bubbling until the pH of the buffer solution reaches 7 4 86 DMI Pressure Myograph System Model 110P amp 111P 25x Concentrated PSS 1000ml Solution 1 Chemical MW g mol Conc mmol l Conc g l NaCl 58 44 118 99 173 850 KCI 74 56 4 69 8 750 CaCl 2H O 147 02 2 50 9 200 Solution 2 Chemical MW g mol Conc mmol l Conc g l MgSO 7H O 246 48 1 17 1 225 KH2PO 136 09 1 18 4 025 Solution 3 Chemical MW g mol Conc mmol l Conc g l EDTA 372 24 0 03 0 250 1 Dissolve the chemicals for solution 1 in about 800ml double distilled HO in a 1000ml graduated bottle Dissolve the chemicals for solutions 2 and 3 in 75ml double distilled HO in individually cylinders Gently heat solution 3 to dissolve the EDTA 2 Solution 2 and 3 is added to solution 1 and the graduated bottle is filled with additional double distilled H O to reach a final volume of 1000 0ml Before use 3 Dilute the 25 x PSS stock solutio
5. tion menu on the Myo Interface e Ifthe value is O or above 6500 then the force transducer is broken and needs to be changed e lf the reading is between 1 499 or 5000 6500 then contact Danish Myo Technology for further instructions If the message OFF is displayed in the force reading line in the Main Menu on the Myo Interface then the force transducer is broken and needs to be replaced In case of any other problems related to the force transducer please contact DMT for further instruction and advice User Manual vers 3 2 Chapter 5 69 70 5 1 7 Checking the Pressure Transducers The Pressure Myograph Unit contains of two pressure transducers P1 and P2 based on Wheatstone bridge circuit The transducers are placed inside the pressure myo graph unit to protect them from mechanical damage The pressure transducers are very vulnerable to applied pressures exceeding 300mm Hg a pressure which is easily exceeded if trying to flush the transducers using a syringe Important DMT recommends that the Myo Interface is connected and turned ON when flushing the pressure transducers tubing and glass cannulas using a syringe This enables an exact control of the pressure applied on the pressure transducers which will prevent them from being damaged The applied flushing pressure should not exceed 150mmHg 5 1 7 1 Simple Pressure Transducer Check 1 Ifthe pressure reading on the Myo Interface appears unstable then first chec
6. 110P is equipped with a Zeiss Axiovert 40 microscope The Zeiss Axiovert 40 is an inverted microscope which provides the highest degree of flexibility in image analysis For instance the Zeiss Axiovert 40 enables the use of a wide range of differ ent objectives and sophisticated fluorescence techniques Contact DMT Sales Department for further product information and prices on any optional equipment for the Zeiss Axiovert 40 DMT Pressure Myograph System Model 110P amp 111P 2 3 Setting up Step by Step Note If you have purchased a computer from DMT in conjunction with your Pressure Myograph System then the Euresys Picolo frame grabber PCI card has already been installed along with drivers and the MyoView software Follow the procedures in Section 2 3 1 to set up the Pressure Myograph System If you have not purchased a computer from DMT follow the procedures in Section 2 3 2 p 20 to install the Euresys Picolo frame grabber PCI card drivers and the Myo View software on your own computer 2 3 1 Setting up the complete Pressure Myograph System This section describes how to connect the cables in the Pressure Myograph System as illustrated in fig 2 1 and fig 2 2 Before connecting any of the myograph equip ment ensure that the frame grabber card is installed into the computer s PCI slot See installation instructions in Section 2 3 2 Note Before proceeding with the connection procedure make sure that the Myo Interface
7. EEE EEN AE EN EEN E AN E EE ES 48 439 1 Loading a Saved SEUUCNCO EEE EE EN 48 isons Eding 00 2 GE EE NE 48 KEENE 50 4 3 5 4 Saving SEUENEE Lande eeknrnnedessneekesle dekade bemannede amnsernn 50 4 3 5 5 Exporting a Sequence as a Jext ME seen 50 EG NE EEE NE EE 50 ROLE NENNE 51 4 3 6 2 Calibration of Image AnalySiS rrwurrrrnnvnnnnnnnnnnnnnnnerernnnnnnnnnnnunnnennvnnnnnnnnnnnnunennnnrevnnnnnnnnnnunnne o1 406 Staring Real ME NN Eve 51 4 3 6 4 Configuration of Vessel Detection and Analysis VDA cccccssssseeeeeeeesssseeeeeeeeaseees 52 4 3 6 5 Logging and Reviewing Results of Analysis rrurrrrrnnnnnnnnnnnnnnnennvrnnnnnnnnnnnnnnnnnnrnvnnnnnnnnnnnnnne 53 4 4 How to Implement an Experimental Protocol rurnnnnennnnnnannnnnnnnnnnnnnnnnnnnnvnnnnvennnnennnr 54 4 5 How to Calibrate the Image Analysis rrrnnnnnrrnnnnnnnrnnnnnnnrnnnnnnnnnnnnnnnrennnnnnrennnnnnnrnnnnn 55 AO HOw BENENE ane 56 TN 58 Chapter 5 Pressure Myograph Manual nanrnavnnnnnnnnnnnnnnnnnnnnnennnnnnnnnnennennnennnn 59 5 1 The Pressure Myograph UNI Cisaiseicsnssienescecsssnatavesarestessientecsesnsvansdaetecanaast anzanrauanerseceeens 59 5 1 1 Adjustment of Glass Cannulas rrrevannavnrnnanannernnnnnnennnnnnnnnnanunnernnnnunnnnnnnnnrnnnnnnnernnnnunennnnnnnennnn 60 DL NNN ENN 61 5 1 3 How to Read a Millimetre Micrometer ccccccececeeeeesseseeeeeeeeeeceeeaeaasseseeeeeeseesseseaaseseeeeeess 62 DLA Force and Pressure Call Need 63 5 1 4 1 Principle
8. Force and Pressure Calibration Section 5 1 4 contains a complete explanation of how to perform the force weight calibration and pressure calibrations As a part of the general maintenance of the myograph DMT recommends that the myograph is force and pressure calibrated at least once every month DMT also recommends that the myograph is force and pres sure Calibrated every time the system has been moved or has not been used for a long period of time 5 1 4 1 Principles of Weight Calibration Weight calibrating the force transducer is based on simple physics the net torque acting on a balance when applying a certain amount of weight The magnitude of the torque T about a point of rotation P is defined by Py t r F sind where r is the distance from the point of rotation to the point on the object where the force F is acting with the angle of 0 Applying the physics to the weight calibration setup is illustrated in Fig 5 8 User Manual vers 3 2 Chapter 5 63 64 Transducer Arm TA Figure 5 8 Thereotical principle of the weight calibration Applying the weight on the pan arm creates a net torque acting at the center of grav ity resulting in a force F acting on the force transducer The following two equations describe the forces working in the weight calibration system Ly TE sole 1 0 im g sind weigt 2 t on where r is the length of the pan arm F is the force acting on the pan arm
9. LCD e Water bath including heater DMT recommendes Julabo 5L e Dissection Petri Dish DMT recommends a 9 cm glass Petri dish coated with a 5mm Sylgaard polymer layer 6 2 Pressure Myograph System Accessories This section contains a list of special accessories available for the Pressure Myograph System Model 110P amp 111P e DMT FlowMeter Model 161FM Item 10720 e pH Probe for Measuring pH in the Superfusion Buffer Standard Needle Probe DMT recommends a Metrohm combined micro glass electrode e pH Probe for Measuring pH in the Perfusion Buffer Flow through Probe e Peristaltic Pump for Superfusion Flow DMI Pressure Myograph System Model 110P amp 111P 6 3 Pressure Myograph System Spare Parts This section contains a complete list of standard available spare parts for the Pres sure Myograph Model 110P amp 111P For parts not listed in this section or for special parts which need to be made customised please contact DMT for further informa tion e Glass Cannulas Item GC 1000 e Suture Wire Item SW 2000 e Calibration Kit Item CK 345110 e Chamber Cover Item CC 1110120 e Force Transducer Item FT 1110120 e Pressure Transducer Item PT 1116115 e High Vacuum Grease to Seal Force Transducer Pin hole Item HVG 1000 e Silicone Tubing at Standard Sizes for the Pressure Myograph System Item ST 1000 User Manual vers 3 2 Chapter 6 19 Chapter 7 Getting Starte Chapter 7 contains some practical pro
10. are adaptive with respect to earlier edge detection of the lumen In other words they use the properties of the vessel determined in the un contracted state to estimate the dimensions in the partially or fully contracted state The incompress ible model assumes that the vessel walls contract without changing their cross sec tional area The extinction model is a unique feature of MyoView which uses an imaging model of the vessel and a measure of the total light absorption of the vessel to estimate the lumen size The user has to determine by experience which model to be used in particular experimental setup Since the analysis is effective in real time the user can make appropriate adjustments whilst viewing their effect until the edge detection is optimal Figure 4 30 Configura tion of vessel detection and analysis MyoView can analyse vessels positioned in either the vertical or horizontal planes The configure analysis dialog box can be used to select the appropriate orientation of vessel default vertical This feature is useful in Some microscope set ups where the vessel appears horizontal in the captured image 4 3 6 5 Logging and Reviewing Results of Analysis The configuration and control of data logging is performed from the data control bar If image analysis is active the derived parameters are passed to the data control bar for display and or logging to file This is also the case when the sequence
11. bar is visible by selecting the sequencer icon in the tool bar Select the popup menu click on h from the sequencer control bar and select the Add entry menu item A dialogue box will appear to enter the details of the sequence Each line of the a sequence i e each entry implements a i E stage of the protocol When you have com ae mill pleted the entry click OK and proceed Duration valt to the next entry You may edit or remove uation i Inflow Pressure g mmHg M entries by appropriate selections from the Outflow Pressure 20 mmHg fy popup menu Temperature 25 dg M Cancel 7 A i 7 When you are satisfied with the entries in ER EN the protocol save the sequence from the Save sequence item on the popup menu A standard dialogue box will appear for file naming and location Figure 4 32 Programming a sequencer event To run the experimental protocol simply click the Start button lon the sequencer control bar The sequence may be started at a given entry by clicking the Start at DMI Pressure Myograph System Model 110P amp 111P button l Both of these actions are also available from the pop up menu click onl on the sequencer control bar and from the main menu Note that if the data logger is not active when a sequence is started MyoView will remind the user to prevent an over sight 4 5 How to Calibrate the Image Analysis The ima
12. complete dissection and mounting procedure Appendices contain additional information about myograph service shipping instruc tions system specifications and fuse changing DMI Pressure Myograph Systems Model 110P amp 111P Unpacking the myograph system Please take a few minutes to carefully inspect your new Pressure Myograph System for damage which may have occurred during handling and shipping If you suspect any kind of damage please contact us immediately and the matter will be pursued as soon as possible If the packing material appears damaged please retain it until a possible claim has been settled We recommend that you store the packing material for any possible future transport of the Pressure Myograph System In case of transport and the original packing mate rial is unavailable please contact DMT Sales Department for advice and packing instructions After unpacking your new Pressure Myograph System please use the following list to check that the system is complete 1 Pressure Myograph Unit e Force calibration kit Including bridge balance and 2 gram weight e Resistance tube shunt Calibration tube e Two glass cannulas e Chamber cover 2 DMT Microscope Model 111P e Olympus Objective Micrometer including Microscope Objective Holder 3 Zeiss Axiovert 40 Inverted Microscope Model 110P e Olympus Objective Micrometer 4 Pressure Myo Interface e Pressure Myograph Unit connection cable wit
13. is performed without further oxygenation of the PSS 4 Pin down the proximal end of the intestine section on the left hand side of the Petri dish without stretching the vessels Pin down the remaining of the intestine section in an anti clockwise direction In this configuration proximal end at the left side distal end at the right side and running anti clockwise from proximal to distal side the feeding vasculature is on the far side of the intestine and the veins are usually uppermost 5 Select the vessel segment to be investigated fig 7 1 First time myograph users are recommended to start dissecting and mounting vessel segments from the first or second branch from the superior mesenteric artery approximate internal diameter 200 300um 80 DMI Pressure Myograph System Model 110P amp 111P Second Branch Figure 7 1 Branching of the mesenteric arteries 6 Use high quality forceps and ocular dissection scissors see Chapter 6 1 to dis sect the vessel segment of interest Start cutting through the mesenteric mem brane along both sides of the vessel about 1 2mm from the vessel To avoid ac cidentally cutting the artery always cut along the length of the vessels and never perpendicular to them fig 7 2A B Figure 7 2 Removal of adipose tissue around the area of interest 7 Dissect away as much adipose tissue as needed around the vessels to distin guish between the artery and vein The artery can easily be identified b
14. myograph unit as illustrated in fig 5 9 4 Make sure that the tip of the transducer arm on the balance is placed behind the glass cannula fixation plate as illustrated in fig 5 10 Carefully move the Figure 5 9 Weight Calibration Set up User Manual vers 3 2 Chapter 5 65 66 calibration bridge until the tip of the transducer arm is placed freely behind the glass cannula fixation plate which means it does not touch the fixation plate Figure 5 10 Illustration of how to fit the balance just behind the glass cannula fixation plate 5 Go to the Setup Menu on the Myo Interface and choose Force to weight calibrate the force transducer Press F4 to start calibration Make sure that absolutely no force is applied on the force transducer by checking that the tip of the transducer arm is not touching the glass cannula fixation plate Also check that the relative force reading in the display is sta ble Press F3 to proceed with calibrating Carefully place the 2 g weight on the pan as illustrated in Fig 5 8 p 64 The force applied on the force transducer should mimic the stretch created by the contraction of a mounted ring preparation Wait until the relative force reading is stable Press F3 to finish the calibration Press Esc and go to the Main Menu on the Myo Inter face The force reading on the Myo Interface should now be very close to 9 81 mN If the force reading is different from 9 81 m
15. of the buffers used to calibrate the pH probe pH measurements are temperature dependent and in the temperature compensation line it is possible to choose be tween an AUTO or MANUAL temperature compensation In the AUTO mode the temperature probe is used to measure the temperature of the calibration buffers The temperature is then automatically used in the pH calibration In the MANUAL mode the temperature of the calibration buff ers is measured with a thermometer and entered manually in the manual temperature line Press F1 to return to the Setup Menu DMI Pressure Myograph System Model 110P amp 111P Chapter 4 MyoView Software Manual 4 1 Introduction to MyoView MyoView provides an interface for studies of small blood vessels mounted the Pressure Myograph Systems Model 110P and 111P MyoView includes analysis and data logging facilities for recording dynamic events Also experimental protocols can be designed implemented and saved for future use The interface is based on customised window display surrounded by five control bars which may be separated from the main window if required _MyoView 1 1 Toolbar buttons ee AR meal HSE Qle el sa le RIO 5 Parameter Control Bar ee mam g Data Con trol Bar e e Te Framegrab ber Control e 8 Analysis Control Bar Sequencer Figure 4 1 MyoView Main Window The control bars located at the bottom and to t
16. on amp the control bar but may also be drawn in the main b window Basic controls of the graphics are supplied axis ranges legend but the display is provided as an experimental monitor rather than a post produc tion facility A large internal data memory is pro vided so that the user can scroll back through the graphical record of the parameters even while the experiment is running The logged files may be read into a standard spreadsheet e g Microsoft Excel High quality graphical reproduction of data fields can then be created from a standard Office applica tion The popup menu on the control bar select h can be used to access all functions and this menu can also be revealed by a right mouse button click when the Windows cursor is above the control bar Figure 4 18 Data Control bar A more detailed description of the functions avail able in the data control bar follows under these sub headings e Selecting Data for Display e Configuration of Data Display e Control of Data Display e Data Logging 4 3 4 1 Selecting Data for Display The data control bar can be used to display up to six parameters in the interface These may be selected from the list box menus Control parameters of the myograph e g pressures or derived parameters e g lumen diameter may be displayed The display has an internal memory of several hours where parameters are stored in that memory even if they are not displayed The memory can be
17. program is protected by copyright law and international treaties Unauthorized reproduction or distribution of this program or any portion of it may result in severe civil and criminal penalties and will be prosecuted to the maximum extent possible under law Cancel Choose Destination Location Ed It IS recomme nded to i nsta Il the soft Setup will install YediView in the following folder ware i n th e d efa u It fo d er To install to this folder click Next To install to a different folder click Browse and select another I 77 folder Click Next You can choose not to install Vediview by clicking Cancel to exit Setup Destination Folder C Program Files Vediview Browse ones Setup Type Ed Cl ick N ext Click the type of Setup you prefer then click Next VediView 1 2 Description VediView 1 2 for Picolo Framegrabber lt Back Cancel User Manual vers 3 2 Chapter 2 23 VediView x Setup will add program icons to the Program Folder listed below You may type a new folder name or select one from the existing Folders list Click Next to continue Click Next Program Folders Existing Folders Administrative Tools Euresys Picolo Industrial Games Java Web Start Printe Internet Printing Startup lt Back Cancel Setup Complete The VediView Software package is now installed VediView 1 2 has been sucessfully instal
18. re selecting the Set analysis rectangle icon El These processes may be repeated until the user is satisfied with the selected region 4 3 6 2 Calibration of Image Analysis Calibration of image dimensional analysis can be done in sensible units e g microns defined by the user To perform such a calibration the user is referred to Section 4 5 p 55 How To Calibrate the Image Analysis The default calibration is performed in units defined by the image data resolution i e pixels MyoView can calculate flow rate and vascular resistance but it requires knowledge of the fluid viscosity and vessel length These values must be entered in the analysis configuration dialogue box accessed from the tool bar icon El or from the popup menu The default viscosity value is the one for water which is suitable for the majority of myograph systems The vessel length can be measured by the user using MyoView or any stereomicroscope equipped with an eyepiece reticule Configure Vessel Detection and Analysis Lumen Detection Physical Parameters Wiscosity Name 1 e O03 Vessel Length mm pr Incompressibility Model Image Orientation Horizontal Vessel Figure 4 23 Configura tion of vessel detection and analysis 4 3 6 3 Starting Real Time Analysis Before starting the image analysis it is necessary to select the analysis rectangle See above If the user wishes to obtain calibrated meas
19. regulator and to the myograph chamber directly 7 Gradually pressurise the mounted vessel from 10 to 40mm Hg over a period of about 15 minutes 8 After the gradually pressurising allow the mounted vessel to equilibrate at an intraluminal pressure of 40mm Hg in a No flow state for about one hour The No flow state is activated in the Pressure Menu on the Myo Interface 9 Following the period of intraluminal pressure equilibration equilibrate the vessel longitudinally using the micropositioner attached on the right glass cannula Zero the force reading on the Myo Interface and adjust the mounted vessel longitudi nally to a point of no slack no force DMI Pressure Myograph System Model 110P amp 111P Figure 7 7 MyoView picture of a mounted vessel User Manual vers 3 2 Chapter 7 7 3 Buffer recipes 7 3 1 Physiological Saline Solution PSS 1x PSS 1000ml Solution 1 Chemical MW g mol Conc mmol l Conc g l NaCl 58 44 118 99 6 954 KCI 14 56 4 69 0 350 MgSO HO 246 48 Tr 0 289 KH PO 136 09 1 18 0 161 Solution 2 Chemical MW g mol Conc mmol l Conc g l CaCl 2H O 147 02 2 50 0 368 Solution 3 Chemical MW g mol Conc mmol l Conc g l NaHCO 84 01 25 00 2 100 EDTA 372 24 0 03 0 010 Glucose 198 77 5 50 1 091 1 Dissolve the chemicals in approximately 100ml double distilled HO as three individual solutions as described in the table above Gently heat solution 3 to
20. superfusion buffer Also the diagrams include the DMT Flowmeter Model 161FM The Flowmeter enables flow measurements in the range of 1 500ul min Contact the DMT Sales Department for further product information and prices on any of the optional products shown in fig 2 1 and fig 2 2 Fig 2 1 and fig 2 2 are intended as an overview for the set up of the Pressure Myo graph Systems followed by a detailed step by step description in the remaining sec tions of Chapter 2 User Manual vers 3 2 Chapter 2 17 18 2 2 Pressure Myograph System Model 110P pH Meter optional Myo Interface Rear Panel Peristaltic Pump optional Connection Cable with temperature probe PC Data Acquisition and Sofware FlowMeter 161FM optional Microscope Pressure and Oxygen Supply Oxygen Superfusion Supply Buffer CED Vaccum Pump optional Sucction Waste Bottle Bottle Inlet Superfusion buffer Outlet Superfusion buffer Inflow Perfusion buffer Outflow Perfusion buffer Pressure Regulator Rear Panel Figure 2 2 Example of the Complete Pressure Myograph System Model 110P The major difference between the Model 111P and the Model 110P is the choice of microscope The Model 111P is equipped with the DMT Microscope which is a stand ard inverted microscope with a built in CCD camera The DMT Microscope is very use ful and recommended in set ups requiring standard image analysis The Model
21. third round grounding pin The wide blade or the third prong is provided for your safety If the provided plug does not fit into your outlet consult an electrician for replacement of the obsolete outlet e Be advised that different operating voltages require the use of different types of line cord and attachment plugs Check the voltage in your area and use the correct type See the table below Line plug according to standard 110 125 V UL817 and CSA C22 2 No 42 220 230 V CEE 7 page VII SR section 107 2 D1 IEC 83 page C4 240 V BS 1363 of 1984 Specification for 13A fused plugs and switched and unswitched socket outlets Protect the power cord from being walked on or pinched particularly at power plugs and the point where they connect to the apparatus Refer all servicing to qualified service personnel Servicing is required when the apparatus has been damaged in any way such as the power supply cord or plug is damaged liquid has spilled onto or objects have fallen into the apparatus the apparatus has been exposed to rain or moisture does not operate normally or has been dropped DMI Pressure Myograph System Model 110P amp 111P EMC EMI This equipment has been tested and found to comply with the limits for a Class B Digital device pursuant to part 15 of the FCC rules These limits are designed to provide reasonable protection against harmful interference in residential installations This equipment generat
22. to control flow and pressure settings 5 4 1 Control of Flow and Pressure Settings The Pressure Regulator consists of two individual circuits an air circuit and a buffer circuit as illustrated in fig 5 15 p 73 The Pressure Regulator transfers the pressure and flow settings from the Myo Interface by individual control of the air pressure in the two buffer bottles Important Never fill the bottle with more than 250 ml buffer as higher volumes makes pre cise control of the pressure more difficult 5 4 2 No Flow Setting Activating the No Flow setting in the Myo Interface closes the No Flow Valve in the diagram shown in Fig 5 15 Closing the No Flow Valve serves to prevent any buffer back flow through the mounted vessel Activating the No Flow setting turns on the No Flow indicator lamp on front of the Pressure Regulator Activating the No Flow setting in the Myo Interface deactivates the P2 pressure transducer and the P1 pressure transducer takes control of the pressure applied on the mounted vessel At the same time the P3 pressure transducer is activated con trolling the air pressure in the left bottle with bufffer in fig 5 15 Activating P3 serves to prevent a huge pressure to build up in the left bottle with buffer which may dam age the mounted vessel when re establishing the flow DMI Pressure Myograph System Model 110P amp 111P Inlet to P1 on Pressure Myograph unit Air In Max 1 At
23. used with a video microscope system in which the magnification settings are changed If for example MyoView is calibration using an x20 microscope objec tive and subsequently an x50 device replaces the objective then the calibration can be adjusted by a suitable factor in this case 2 5 Given the optical factors affecting such a change it is always worth checking the calibration for obtaining the most accurate results 4 6 How to Customise the Interface The functionality of MyoView is based on a series of Control Bars that may be configured in many different ways The interface can be changed to suit a particular experiment or user Once the interface has been designed it may be saved for any future use The default interface setup Setup 0 is initially set to the factory default values Although set ups can be redefined the factory defaults can never be changed A custom interface may be designed and implemented by following the procedure 1 Launch MyoView and select the User Setup from the view menu If this is the first time you are using MyoView then the interface set up will be equal to the factory setting Note that you can always return to the factory settings by selecting factory settings from the options in the user setup L kapi 1 1 ore amras Fiane Giebber Daa Sequence rake Helo alal xj Tami wnan pawe g uenig po fae ge Durem pao a dg Fws f
24. valve will make it very difficult to control the pres sure and make the pressure readings unreliable 5 1 5 Replacement of Glass Cannulas The mounted glass cannulas are easily replaced with new ones by carefully loosen ing the two screws F and G in fig 5 2 p 60 The recommended size for the new glass cannulas is 16mm for the glass cannula connected to P1 right side and 14mm for the glass cannula connected to P2 left side 5 1 6 Checking the Force Transducer The myograph force transducer is a strain gauge connected in a Wheatstone bridge The force transducer is placed in a separate compartment on top of the pressure myograph unit The separate compartment provides some mechanical protection for the force transducer but the transducer is still very vulnerable to applied forces exceeding 1 newton 100 gram or fluid running into the transducer compartment due to insufficient greasing of the transducer pinhole This chapter describes how to check the force transducer for any kind of damage 5 1 6 1 Simple Force Transducer Check 1 If the force reading on the Myo Interface appears unstable then first check that the Myo Interface and the Myograph Unit are properly connected through the 25 pin grey cable 2 If the force reading still appears unstable then perform a new weight calibration of the fore transducer as described in Section 5 1 4 2 During the weight calibration monitor the relative force reading values in the calibra
25. vers 3 2 Chapter 4 37 38 Sequencer Control Bar is used to programme various sequences of pressure para digms to be tested in a particular experiment protocol Sequencer gt mf 0 fra 0 Initialise protocol 1 MN 20 ER 1 Increase inflow 1 DD W 0 Ww 20 Cur 2 59 2 Increase inflow 0 40 OW 20 W 20 gt 3 Wait for 1 minute B0 40 20 el 4 Operator interve 1 40 20 20 5 Waitfor minutes 120 40 20 20 E Retumto initial 1 Figure 4 6 Sequencer Control Bar Analysis Control Bar is used to define and control the image processing functions and display results Image Analysis Figure 4 7 Analysis Control Bar DMT Pressure Myograph System Model 110P amp 111P 4 2 3 Toolbar MyoView contains a dockable tool bar containing bottoms with specific icons used as Short cuts to various MyoView operations i e in the menus and control bars The tool bar appears in the top of the interface underneath the menus Some users find the iconic representation more convenient when they have become more familiar with the software i _ HSE see Flue BJ GE Figure 4 8 MyoView Toolbar By passing the pointer over each icon and pausing a tool tip will remind the user of the function of each icon Tool tips are implemented throughout MyoView The meaning action of each icon is described in fig 4 9 ES Turn on off Frame Grabber Control Bar
26. when applying the weight F is equal acceleration of gravity times the mass of the weight r is the length of the transducer arm and F is the force acting on the force trans ducer The net torque acting at center of gravity is constant for the weight calibration setup which makes equation 1 and 2 equal making it possible to calculate the force acting on the force transducer fr Sine Fon OM cig 8 SING FL fa ag 8 sind E SINE DMT Pressure Myograph System Model 110P amp 111P As the length of the pan arm is 2cm the length of the transducer arm is 4cm the weight is 2g both angles are 90 and the acceleration of gravity is 9 81ms gt the force acting on the force transducer is F 2cm 2 gram 9 81ms sin90 4cm SiIN90 FEN As 1N is equal 1kg m s F is equal to F 9 81mN 5 1 4 2 Weight Calibration Procedure The section contains a complete step by step description of how to weight calibrate the force transducer 1 Fill the myograph chamber with double distilled water and move the glass can nulas apart 2 Place the calibration bridge balance and weight on the myograph unit allowing it to be pre warmed together with the myograph unit Turn on the heating in the Main Menu on the Myo Interface 3 After approximately 20 30 minutes the whole system will have reached the target temperature normally 37 C Place the warmed calibration bridge and balance on the
27. 1 inflow to 20mmHg and P2 outflow to 5mmHg If you connect the tubing to the myograph you first need to fill the tubing to pre vent air bubbles this will happen by the pressure difference Start with turning status ON the tubes will fill up The two tubes are interconnected connect the end of it to P2 on the myograph and disconnect the tubes halfway the end of the P2 tube must be connected to P2 on the pump unit The tube originating from the P1 bottle is to be connected to P1 DMI Pressure Myograph System Model 110P amp 111P During this both 3 way valves remain closed just open them to rinse through the second outlet on the myograph but make sure you don t open the vault in a way it will open directly to the artery Now turn OFF status and reduce pressure for P1 to 5mmHg During this process release pressure by removing the 5 tube from the pump unit and make sure at the end there is no pressure Make sure for both inlet and outlet the pressure is 5mmHg and that the flow and Status are off before you open the valves After opening both valves turn status ON Increase pressure by 10mmHg about every 5 minutes until 7OmmbHg this can be done in Myoview If there is a leak P2 outflow will be too low red cross check the tubing and ar tery If there are branches holes mount artery in a different way Software Get the artery in the main window of Myoview and draw a square over it Calibrate to 10x diameter file using x but
28. 4 13 Turning off the analysis control bar in the view menu Once a user has designed and configured the appearance of the interface the setting may be saved and subsequently loaded The default setup Setup 0 is initially set to the factory defaults however the user may change this at any time When MyoView is closed the set ups remain in memory and may be recalled The factory defaults may be restored at any time and can never be changed by the user 4 3 2 Parameter Fundamental settings of the Pressure Myograph System such as force pressure and temperature can be monitored and controlled from the para Target meter control bar Manual control of the settings Temp deg C 21 00 25 00 E3 is also obtainable via edit boxes on the control Inflow mmHg 0 00 20 00 EJ bar or an automatic sequence of settings can be implemented by using the sequencer feature Outflow mmHg 20 00 20 00 Output from the force transducer in the Pressure Force mN li a0 Myograph System is also displayed on this control bar The popup menu on the control bar select XJ Figure 4 14 Parameter Control bar Parameter DMI Pressure Myograph Systems Model 110P amp 111P can be used to edit the target settings and to access all other functions of the control bar This popup menu can also be revealed by a right mouse button click when the Windows cursor is above the control bar Hardware control may be switched on or off by mean
29. Can I turn these off The log files myo contain entries for all possible parameters Some parameters will not have been calculated because of the particular MyoView settings for a given experiment and therefore no data will appear in those columns This feature is to ensure that data are not inadvertently lost by not selecting that data for the log file It further protects against having to repeat experiments for which a new requirement for data only emerges after the experiments are completed I am performing an experiment but the data history graph is not changing The data history graph pauses whenever the pause button is pressed or whenever the user scrolls through the recorded data To un pause the graph press the pause resume button How do I pause the data logging to disk The data logging process can be paused or resumed with the start stop data logging button MyoView will continue to store data even when data logging is suspended and that data can be viewed using the data history graph DMI Pressure Myograph System Model 110P amp 111P Chapter 5 Pressure Myograph Manual Chapter 5 contains a complete explanation to all mechanical features of the Pressure Myograph System This includes how to use the DMT Microscope and how to use adjust and separate the Pressure Myograph Unit 5 1 The Pressure Myograph Unit The pressure myograph unit is placed on the DMT microscope as illustrated in fig 5 1 Press the
30. D 0 0 Pressure Transducer and Glass Cana S svarene 15 5 5 4 Pressure Regulator CICA ING uanmeldt ipen ammeeinmnneietn 15 55 5 Cleaning Pressure Myograph CHEMI GM Larver aeaa ierden 16 Chapter 6 Myograph Accessories and Spare Parts urnnnnnunnnnnnnnnnnnnnnnnannanenn 78 6 1 General Myograph Equipment ssavenanvnnnnsunnnnnnnnennnvnvnnnnsnnnnnnnnnnnnnennnvnnnnnnsnnnnunennnnunene 18 6 2 Pressure Myograph System Accessories rsassunvnsuunvnnsunnnnnunnnnnnunnnnnnnnnnuunnnnuunennnnunene 18 6 3 Pressure Mograph System Spare PAINS svare 19 DMI Pressure Myograph System Model 110P amp 111P Chapter 7 Getting Started arrnnrnannnnnnnunnnnnnnnnannnnnnnnnnnnnnnennnnnnennnennennnennennnennn 80 7 1 Dissection Protocol for Small Mesenteric Arteries srrrvasunrrnannnennnnnnnnsnnnnannnnvnsnne 80 1 2 MOUNTING Protocol for Small EE aan 84 Appendix 1 Pressure Myograph Set up rar auunnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnennnnnnunn 88 Appendix 2 Terms Of Warranty aareaaxenannnannnannnnnnnnnennnennnnnnnnnnnnnnnevnnevnnnnnnennnnn 91 Appendix 3 Service Check nnnnnnnnnnnnnnnnnnnnnnnnnnunnnnnnunnnnnnnnennnnnnennennnennnnnnennnnnnen 92 Appendix 4 Shipping Instructions anrnanvnnnnannnnnnnnnnnnnnnennnnnnennnnnnennnennennnennnnn 93 Appendix 5 System Specifications anrenannnnnnnnnnnnnnnnnennnennnnnnnnnnnennnevnnennnennnnn 94 Appendix 6 Fuse Changing sanvnannnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnennnnnnennennnennnnnnennnnnnen 95 User
31. LO frame grabber The frame grabber control bar is used to configure the current status of the video acquisition and to manipulate the display of frames in the interface The real time frame grabbing mode may be switched off when not in use or when the user wishes to save a frame to file for future reference A zoom in out facility is also provided The popup menu on the control bar select x can be used to access all functions of Figure 4 14 Framegrabber Control the frame grabber bar This popup menu can also bar be revealed by a right mouse button click when the Windows cursor is above the control bar A more detailed description of the functions is available in the frame grabber control bar follows under these sub headings e Displaying Images e Image Calibration e Zooming e Saving an Image e Frame Grabber Configuration e Distance Measurement Note When configuring the camera and frame grabber it is important to ensure that the image is NOT saturated as this may affect the quality of the edge detection analy sis The presence of completely white sections within the image indicates a saturation condition which may be rectified by either reducing the light source intensity or reduc ing the camera gain using the frame grabber configuration 4 3 3 1 Displaying Images The real time framegrabbing mode may be switched on or off by means of the Start stop display button or from the popup menu activated click on f
32. MT Microscope Camera Objective Adjustment Light source Serial interface Voltage Pressure Regulator Pressure range Pressure calibration Fluid capacity Gassing Heating Valves Voltage gt 60um Manually X Y amp Z Glass pipettes Single Acid resistant stainless steel Max 10 ml 3ml typical With built in pipes for suction gassing Superfusion 50 mN 0 01 mN 0 250mm Hg 1 0mm Hg Build in Ambient 500C 0 10C Yes included external Manual Manual 1 0V F S Serial interface RS232 RS485 115 230V 50 60 Hz Watec 1 3 bw ccd 10x Zeiss CP achromat X y amp z directions Red light built in RS232 115 240 V 50 60Hz O 250mmHg requires external pressure source Manual 250 ml bottles Built in Built in Safety and reduction 115 230V 50 60 Hz DMT Pressure Myograph System Model 110P amp 111P Appendix 6 Fuse Changing gt gt The main fuse of the myograph system is placed inside the power inlet on the Myo In terface If the fuse blows it is easily changed using the following procedure Important If the fuse needs to be changed make sure that the replacement fuse is equal to the one blown Specifications T1 6A 250V 6 3 x 32mm Use a small screwdriver to open the voltage selector block User Manual vers 3 2 Appendix 6 95 rot SRC T eS Ck 096 DMT A S Skejbyparken 152 DK 8200 Aarhus N Denmark
33. Manual vers 3 2 Contents 10 About this manual This manual contains a complete list of procedures describing how to install maintain and get started using the Pressure Myograph System Model 110P amp 111P Version 3 2 Chapter 1 provides an overview of the construction and basic features of the com plete Pressure Myograph System Chapter 2 describes step by step how to set up a complete 110P or 111P Pressure Myograph System including all kind of accessories Chapter 3 is a complete manual to the Myo Interface The chapter describes in detail the construction of the menu system and how to use all the features of the Pressure Myograph System Chapter 4 is a complete manual to the MyoView Pressure Myograph software The chapter describes in detail the construction of the menu system and how to use all the features of the Pressure Myograph System Chapter 5 is a complete manual to the Pressure Myograph System The chapter describes in detail how to use the DMT microscope how to use and adjust the pres sure myograph chamber and finally instructions to daily maintenance of the Pressure Myograph System Chapter 6 is a list of equipment needed to set up a complete Pressure Myograph System The chapter also contains a list of accessories and spare parts available to the Pressure Myograph System Model 110P amp 111P Chapter 7 contain procedures describing how to get started using the Pressure Myo graph System This includes a
34. N then try to cali brate the fore transducer once again starting with step 3 9 After calibrating the force transducer carefully remove weight balance and calibration bridge The Pressure Myograph System is now ready for use longitudinal force measurements DMT Pressure Myograph System Model 110P amp 111P Setup Menu Calibrate Press Resovoir STANG Option 2 F4 Calibrate Force Apply no Force Push F3 when stable Force 1938 F3 Calibrate Force Place 2 g on the pan Push F3 when stable Force 2245 F3 Calibrate Force DONE Push Esc Force Zero 20315 Force gain 668 3 5 1 4 3 Pressure Calibration Procedure The setup to perform the pressure transducer calibration procedure is illustrated in Fig 5 11 Carefully follow the procedure described below to pressure calibrate the Pressure Myograph System Figure 5 11 Illustration of set up for pressure transducer calibration E Connect the Pressure Myograph Unit to the Pressure Regulator as to perform a standard pressure myograph experiment as illustrated in fig 5 11 Connect the Big Ben Pressure Manometer to the 3 way valve at P1 on the Pres sure Myograph Unit as illustrated in fig 5 11 Inside the myograph chamber on the right side is a small stainless steel pipe which via a small silicone tube is connected to the right glass cannula Fig 5 12 B Carefully disconnect the silicone tube from the stainless steel pipe Carefully discon
35. P2 and 1 on the Pressure Regulator follow the procedure described in Section 5 1 4 3 Important Be very careful not to damage pressure transducer P3 when flushing the buffer cir cuit There are no readings from P3 visible on the Myo Interface Danish Myo Techno logy therefore recommends that the Pressure Myograph Unit always be flushed first In this way it is possible to get a feeling of how much flushing pressure that is equal to 150mm Hg Important II Never flush the air circuits port no 2 3 and 4 on the pressure regulator with any kind of fluid User Manual vers 3 2 Chapter 5 75 5 5 5 Cleaning Pressure Myograph Chamber DMIT strongly recommends that the myograph chamber and surroundings be cleaned after each experiment After a normal experiment use the following procedure to clean the myograph cham ber and supports 1 Fill the myograph chamber to the edge with an 8 acetic acid solution and allow it to stand for a few minutes to dissolve calcium deposits and other salt build up Use a swab stick to mechanically clean all chamber surfaces 2 Remove the acetic acid and wash the myograph chamber and glass cannulas several times with double distilled water 3 If any kind of hydrophobic reagent have been used which might be difficult to remove using step 1 and 2 then try incubating the chamber and glass cannulas with 96 ethanol or a weak detergent solution e g Treepol 4 To remove more resistant or t
36. PCI card as described in the computer or motherboard manual 2 Turn on the Pressure Myograph System and the computer Typically the Windows software will automatically detect the installed frame grabber and starts up a Guide to install new hardware Skip the automatic installation press Can cel and proceed with step 3 3 Insert the MyoView Version 1 2P CD ROM into the computer s CD ROM drive Normally the computer automatically starts installing the software programs If this does not happen run the program setup exe from the CD ROM DMI Pressure Myograph System Model 110P amp 111P 2 3 2 1 Picolo Frame Grabber Driver Installation The auto run will start by installing the drivers for the Picolo Frame Grabber InstallShield Wizard Click Next Picolo Industrial drivers setup Welcome to the Picolo Industrial setup utility coca Warning Ignore the warning In order to use some of the sample programs Click OK WOU need to install the Euresys evision library IF you d like to use Borland C Builder or Delphi sample programs you must install eVision Actives controls To do so you should select the Microsoft Visual Basic Setup type for evision InstallShield Wizard It is recommended to install the soft Choose Destination Location Wa re in the defa u It folder Select folder where Setup will install files Setup will install Picolo Industrial in the following folder Cl
37. Perfusion Buffer Inlet and Outlet Pipes Pressure Transducer Calibration Valve emnene Pressure Valve Activity Indicators Figure 1 5 Pressure Regulator Front Main Pressure Inlet e 115 230 V 50 60 Hz Automatic Voltage Connection Port for the Pressure Myo Interface Selector amp 0 T E l Interface Pressure inlet ON OFF Switch Sa Inlet Pressure Manometer Power Connector e Figure 1 6 Pressure Regulator Back DMT Pressure Myograph System Model 110P amp 111P Chapter 2 Setting up 2 1 Pressure Myograph System Model 111P pH Meter optional Peristaltic Pump optional mm ul Myo Interface Rear Panel Connection Cable with temperature probe PC Data Acquisition and Sofware RS 232 USB FlowMeter 161FM optional Pressure and Oxygen Supply O A DMT Microscope E Oxygen Superfusion g Supply Buffer A q se Sucction gt am Bottle Vaccum Pump optional Waste Bottle Outlet Superfusion buffer Inlet Superfusion buffer Inflow Perfusion buffer Outflow Perfusion buffer Pressure Regulator Rear Panel Figure 2 1 Example of the Complete Pressure Myograph System Model 111P Fig 2 1 and fig 2 2 p 18 are diagrams of complete set ups for the Pressure Myo graph Systems Model 111P and 110P The diagrams include optional equipment such as a peristaltic pump to enable superfusion flow and pH probes for measuring pH in the
38. Pressure Myograph System Model 110P amp 111P J oo J User Manual Version 3 2 DANISA MYO T HmnNunNoBodu 0G DrNnISsH M 0d TE atimnuNOEOL0G DMT Pressure Myograph System Model 110P amp 111P Pressure Myograph System Model 110P amp 111P User Manual Trademarks Pentium is a registered trademark of the Intel Corporation Windows Windows 95 Windows 98 Windows ME Windows NT Windows 2000 and Windows XP are registered trademarks of Microsoft Corporation All other trademarks are the properties of their respective owners DMT reserves the right to alter specifications as required This document was as far as possible accurate at the time of printing Changes may have been made to the software and hardware it describes since then New information may be supplied separately This documentation is provided with the DMT Pressure Myograph System Model 110P amp 111P Version 3 2 Document Number 110P 111P 001A No part of this document may be reproduced by any means without the prior written permission of DMT Copyright 2006 DMT A S M DANISA MYO TECHNOL COG DMT A S DMT Asia DMT USA Inc Skejbyparken 152 Everwin Gardens 420 Beach Rd 204 DK 8200 Aarhus N Rm 502 Block B Sarasota FL 34242 Denmark 521 Wanping Nan Lu USA Shanghai 200030 Tel 45 87 41 11 00 China Tel 1 770 612 8014 Fax 45 87 411101 Tel 86 0 21 64869685 Fax 1 678 302 7013 Fax 86 0 21 64280591
39. alid vessel but only has the outer edges de termined by edge detection MyoView will indicate that an estimation algorithm is being used to determine the inner or lumen edges see fig 4 28 and fig 4 29 This condition will be clear from the analysis results window showing the green model icon Frame Grabber Image Analysis Figure 4 28 One edge detection Figure 4 29 One edge analysis DMT Pressure Myograph System Model 110P amp 111P In this particular example MyoView assumes that since the analysis rectangle con tains of one wall a model is required to determine the position of the inner edges or lumen This assumption is made to cope with images of contracting vessels There are two possible models for this condition and the user may select which to use This choice can be made from the configuration dialogue box which is accessed by selecting the configure analysis icon Ei on the tool bar or select the configure analysis menu item from the popup menu click on x on the analysis control bar Configure Wessel Detection and Analysis Lumen Detection Physical Parameters Viscosity Name fi 003 Vessel Length mm Incompressibility Model Image Unentation Horizontal Vessel The two models available for estimating the lumen dimensions when edge detection is inaccurate or impossible are the incompressible and the extinction models Both models
40. aling up the transducer hole be changed at least once a week 2 DMT takes no responsibilities for the use of any other kinds of high vacuum grease than the one to be purchased from DMT 3 DMT takes no responsibilities for any kind of damage applied to the force trans ducers 5 5 3 Pressure Transducer and Glass Cannulas To prevent the pressure transducers and the tubing inside the pressure myograph unit from being blocked by buffer salt deposits after an experiment DMT recom mends that the system be flushed using the following procedure 1 Disconnect the Pressure Regulator from the Pressure Myograph Unit Keep the Pressure Myograph Unit connected to the Myo Interface to enable continuous monitoring of the pressure readings from P1 and P2 2 Filla small syringe with 8 acetic acid and flush both pressure transducers and glass cannulas by connecting the syringe needle to the 3 way valve for the indi vidual pressure transducers and glass cannulas using a small piece of silicone tube Important Be very careful not to apply a too high pressure when flushing as this may dam age the pressure transducers Keep an eye on the pressure transducer readings on the Myo Interface when flushing and make sure that the pressure does not exceed 150mm Hg 3 Flush the pressure transducers and glass cannulas as described in step 2 Re peat the procedure two to three times 5 5 4 Pressure Regulator Cleaning To clean the buffer circuit port no
41. alues and tolerance settings default values 0 1 for each target value can be set from the popup menu click on h on the control bar or from the main menu To enable maximum flexibility pressure and temperature control may be independently disabled Pressure control is active as long as the hardware control is enabled Temperature control enable disable is available from the popup menu click on on the control bar or from the main menu 4 3 2 3 Automatic Control of Pressure and Temperature The pressure and temperature target settings can be set and re set automatically by means of the sequencer control bar This control is part of the MyoView facility to implement experimental protocols 4 3 2 4 Configure Computer Communications The myograph interface connects to the serial port of the host computer Users may Configure Serial Communications 3 have a number of peripherals connected to COM 1 Comm Fort serial ports on their computer and each will have a numerical identification The appro priate communication port for MyoView can be selected from the Configure Serial Communications icon in the popup menu Figure 4 15 Configuration of serial commu on the parameter control bar After selecting nication this menu item a dialogue box will appear to select the appropriate port default COM1 User Manual vers 3 2 Chapter 4 43 44 4 3 3 Frame Grabber The MyoView software is fully integrated with a TT PICO
42. ay be docked undocked resized or removed altogether from the interface a user may design their own configuration which suits a particular application The MyoView software will perform the same functions irrespective of whether control bars are visible docked or undocked Therefore it is recommended that a new user employs the factory defaults for the interface until they are experienced enough to produce custom settings which may enhance their overall use of the software DMI Pressure Myograph System Model 110P amp 111P There are five Control Bars in MyoView Parameter Control Bar used to control the pres sure and temperature settings of the Pressure Myograph Target values may be entered via the Target menu on the control bar Also the values can be Temp deg C 21 00 25 00 E3 controlled via the Myo Interface and the Myo View sequencer feature Inflow mmHg 0 00 20 00 E Outflow mmHg 20 00 20 00 Force mM 1 00 Figure 4 3 Parameter Control Bar Frame Grabber Control Bar used to control the Frame Grabbe video data acquisition and the display of frames Data Control Bar used to display current and his torical data as well as log data to file Data F fInflov Pressure mmHg Outflow Pressure mm Mean pressure mmHg Target Inflow Pressure Target Outflow Pressur Temperature deg C r Figure 4 5 Data Control Bar User Manual
43. be saved as a text file so that the user has a record of the experimental protocol This text file is created by saving a se quence in the usual way except that when the save dialogue box is active the user will be able to choose to save a seq file for future use in MyoView or a txt file for a record of the protocol The exported file readable by any text editor or standard office application 4 3 6 Analysis The analysis control bar is used to activate and determine the type of analysis to be per formed on the acquired images The purpose p of this analysis is to determine the dimensions me of vessels in the myograph and to derive other parameters e g vessel cross section based upon those dimensions Although the video microscope supplies the images for this analy sis some judgements must be made concern ing the region for analysis and the nature of Figure 4 22 Analysis control bar the analysis MyoView uses a unique vessel detection and analysis VDA methodology to recognise the presence of a vessel and locate its edges This method is largely automatic and effectively checks the correct imaging of the vessel and is capable of estimating the position of lumen edges even when the vessel is contracting when conventional edge detection can not be used Since the analysis is implemented in real time on live images the user can interact with and re configure the analysis so that an optimum setting can be achieved for the giv
44. black lever to lock the Pressure Myograph Unit on the DMT Microscope Figure 5 1 Model 111P Microscope setup User Manual vers 3 2 Chapter 5 59 60 5 1 1 Adjustment of Glass Cannulas Adjustment of the glass cannulas is divided into a pre experimental alignment and general adjustments done during mounting of a blood vessel or when running an experiment General Adjustments General adjustments are performed using the two micropositioners on top of the Pressure Myograph Unit The vertical micropositioner fig 5 2 B is used to adjust the vertical position of the right glass cannula and thereby the vertical position of the mounted blood vessel The horizontal micropositioner fig 5 2 C is used to define the length between the two glass cannulas and thereby the horizontal stretch of the mounted blood vessel A p ev me O Figure 5 2 Adjustment of glass cannulas DMT Pressure Myograph System Model 110P amp 111P Pre Experimental Alignment To make the general adjustments as easy as possible during an experiment it is important to make an alignment of the glass cannulas prior to the mounting of the blood vessel Left Glass Cannula Horizontal positioning of the left glass cannula is adjusted by carefully loosening screw D fig 5 2 Vertical positioning of the left glass cannula is adjusted by care fully loosening screw E fig 5 2 Longitudinal positioning of the left glass cannula is adjusted by gently l
45. djustment Screw e Micropositioner for Vertical Adjustment of Glass Cannulas Force Transducer ev mesmo i Outlet Pressure 25 Pin Serial Port for Commu Transducer Connection Pipe nication with Myo Interface Outlet Valve PO Inlet Valve R i i e Outlet Pressure Transducer Inlet Pressure PEER AEE e Transducer Force Transducer Pin eeeeeereereeee Micropositioner for Longitudinal Adjustment of Glass Cannulas Fixation Plate for Left Glass Cannula 9 Fixation Plate for Right Glass Cannula bd Outlet Connecting Pipe prrseeesssns e Perfusion Inlet Pipe Q r Silicone Tube connecting Outlet Con Silicone Tube connecting Perfusion necting Pipe with Left Glass Cannula i i Inlet Pipe and Right Glass Cannula Left Glass Cannula PE ees NN r Right Glass Cannula cosocecocosocoococosoococococosoccosossooococoocoooooes o Myo Chamber Window Figure 1 1 The Pressure Myograph Unit User Manual vers 3 2 Chapter 1 13 14 1 2 DMT Microscope Model 111P Light intensity regulation plug Lever to fasten Pressure See Myograph Unit fig 1 1 Plug for connec Infrared light scource e Zeiss Achromat 10X XY Z level regulation tion of tempera 0 25 objective of objective focus ture probe Figure 1 2 The DMT Microscope DMT Pressure Myograph System Model 110P amp 111P 1 3 Pressure Myo Interface Up a
46. e These programs are provided to you free of charge as such You should not assume that these programs are error free or suitable for any purpose whatsoever Do you accept all terms of the preceding Disclaimer If you choose NO Setup will close To install the sample programs you must accept this disclaimer InstallShield InstallShield Wizard The Picolo Frame Grabber is now in Stalling from the CD Setup Status Picolo Industrial Setup is performing the requested operations Installing C Program FilestE uresys Picolo Industrial Doct Picolo32 cnt 90 Cancel InstallShield Wizard The Picolo Frame Grabber is now InstallShield Wizard Complete installed Setup has finished installing Picolo Industrial on your computer Click Finish DMT Pressure Myograph System Model 110P amp 111P 2 3 2 2 MyoView Software Installation Finally the installation program automatically continues to install the MyoView data acquisition and analysis software Welcome x The installation will now proceed with Welcome to the VedVien Setup progam This installation of the VediView Software rogram will install VediView on your computer snes Package It is strongly recommended that you exit all Windows programs before running this Setup program CI i k P N 7 Click Cancel to quit Setup and then close any programs you IC ext have running Click Next to continue with the Setup program WARNING This
47. e Gently pull away adipose or connective tissue to make the connective tissue membrane become visible Cut the membrane to remove the tissue 10 Cut the distal end of the artery section to be investigated Afterwards cut the proximal end while ensuring that the vessel segment has the correct length fig 1 5A C DMT Pressure Myograph System Model 110P amp 111P Figure 7 5 A B and C Cutting free the artery to be studied User Manual vers 3 2 Chapter 7 83 84 7 2 Mounting protocol for small arteries This section shortly describes the basic technique of mounting small vessels in the Pressure Myograph Figure 7 6 Mouting of vessel in the Pressure Myograph Chamber 1 Fill the Pressure Myograph Chamber with cold aerated PSS and transfer the iso lated vessel segment to the Pressure Myograph Chamber 2 Carefully cannulate end of the vessel on the right side glass cannula connected to P1 Gently secure the vessel with two fine nylon sutures 3 Gently flush the lumen of the mounted vessel with PSS to remove any blood or debris 4 Carefully cannulate the other end of the vessel with two fine nylon sutures 5 Turn on the heating of the Pressure Myograph Chamber in the Main Menu on the Myo Interface The PSS in the myograph chamber is then gradually warmed to 37 C over a period of 15 minutes 6 Place the chamber cover over the Pressure Myograph Chamber and open for the oxygen supply both to the pressure
48. ectronic and related apparatus for household and similar general USE EN 50 081 1 Electromagnetic compatibility Generic emission standard Part 1 Residential commercial and light industry EN 50 082 1 Electromagnetic compatibility Generic immunity standard Part 1 Residential commercial and light industry With reference to regulations in the following directives 73 23 EEC 89 336 EEC DMI Pressure Myograph System Model 110P amp 111P Contents HAAGCMAKS sessin aa Kaaa aa REEERE ARa 3 SUN Le 4 EMC EN 5 PAPE OV ANS EEE NE 5 Certificate of Conformity arnnnnwnnnnnnnnnnnnnannnnnnnannnannnnnnvnnnnnnnnnnnnnnnnnnvennvennnennnennnene 6 About this Manual cssntsiccnniiensinsnencansetewnsdecsennccanssstwancandeussensnensdadataeninmesecnnmusneuniets 10 Unpacking the myograph System anrnanvnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnennnennennnennennnennennn 11 Chapter 1 System overview r annnnannnnnnnnnnunnnnnnannnnnnnnnnnnnnnennnnnnennnnnnennnennennnennnn 13 ETE ENN 13 1 2 DMT Microscope Model T11P Liuisssarssessasnssanssnssenendannnandnvannsendnendsamtesnnendvedaedad nandaennid 14 1 3 Pressure Myo lnterface wcsennseuesistucse densa daasns cpnnsliacinmdcasaiedehesioantivacaaweructiavnd cainetsodeeaddeees 15 tA POS SUC eS UNCON EE EE 16 Chapter 2 Setting UP se sasisceeccsecsnsstcceasesamnsssssenandansnemnas cstwaasntnesdagnaneenendennaenaduncane 17 2 1 Pressure Myograph System Model 111P vusnnssssaeamesen amts 17 2 2 Pressure Myo
49. en vessel and microscope being used MyoView provides this feature to the user without them requiring any specialist knowledge of the image processing algorithms being implemented The popup menu on the control bar select can be used to access all functions of the analysis bar This popup menu can also be revealed by a right mouse button click when the Windows cursor is above the control bar A more detailed description of the functions available in the analysis control bar now follows under the these sub headings Image Analysis DMI Pressure Myograph System Model 110P amp 111P e Setting the Analysis Rectangle Calibration of Image Analysis e Starting Real Time Analysis Configuration of Vessel Detection and Analysis VAD Logging and Reviewing Results of Analysis 4 3 6 1 Setting the Analysis Rectangle If the user requires a calibrated image analysis the dimensions of the image must be determined before commencing analysis To perform this calibration the user is referred to Section 4 5 How To Calibrate the Image Analysis It is now assumed that the calibration if required has already been performed Before selecting a region for analysis the image should be displayed in the main window Drag and click the mouse to select a rectangle in the main window and then select the Set analysis rectangle icon Ellon the tool bar The analysis rectangle may be re selected by sim ply drawing a second rectangle and
50. es uses and can radiate radio frequency energy and if not installed and used in accordance with the instructions may cause harmful interference to radio communications However there is no guarantee that interference will not occur in a particular installation If this equipment does cause harmful interference to radio or television reception which can be determined by monitoring the interference while turning the equipment off and on the user is encouraged to correct the interference by one or more of the following measures e Reorient or relocate the receiving antenna e Increase the separation between the equipment and receiver e Connect the equipment into an outlet on a circuit different to that which the receiver is connected to e Consult the dealer or an experienced radio TV technician for help Approvals Complies with the EMC standards EMC 89 336 EEC EN 50 081 1 and EN 50 082 1 FCC part 15 Class B CISPR 22 Class B Certified with the safety standards EN 60 065 IEC 60065 Complies with the safety standards UL6500 CSA E65 User Manual vers 3 2 Safety Certificate of Conformity DMT A S Skejbyparken 152 8200 Aarhus N Denmark hereby declares its responsibility that the following product Pressure Myograph System Model 110P amp 111P Version 3 2 is covered by this certificate and marked with CE label conforms with the following Standards EN 60 065 IEC 65 Safety requirements for mains operated El
51. evealed by a right mouse button click when the Windows cursor is above the control bar The easiest way to learn the features of the sequencer is to load a test sequence test seq supplied with the MyoView software Follow the sub headings below that constitute a tutorial on how to load edit run and save sequences e Loading a Saved Sequence e Editing a Sequence e Executing a Sequence e Saving a Sequence for Future Use e Exporting a Sequence as a Text File 4 3 5 1 Loading a Saved Sequence A sequence file seq can be loaded from the pop up menu click on Hi on the sequencer control bar If you have not created your own sequencer files use the sup plied test seq file Select Load sequence from the menu options and a dialogue box will appear to locate and load the chosen file These files are created to execute an experimental protocol and therefore constitute a record of the protocol You may now wish to edit a given sequence 4 3 5 2 Editing a Sequence Once a sequence is loaded into MyoView it may be edited This action will not change the saved version For the purposes of this tutorial it is assumed that the factory supplied sequence test seq has been loaded Each line of the sequence is an entry and constitutes a set of actions for the P110 An entry can be selected by clicking the mouse on the blue LED icon I adjacent to each entry The chosen entry may be then be edited by selecting the Edit entr
52. ge processing functions in MyoView can be Warning Warning datalogger not started Figure 4 33 Datalogger activity warning performed in units derived from the image data i e pixels or can be defined by means of a calibration This involves imaging of an object of known dimensions such as a calibrated by the following procedure reticule The image analysis is 1 Setup the complete Pressure Myograph System to image an object of known di mensions Make sure the framegrabber control bar is visible select the frameg rabber icon al in the tool bar and appears in the main window Make sure the frame grabber is in live mode from the frame gra bber control bar 2 Using the live video setting image the object in the main window and use the zoom facility to obtain the most accurate view of 3 Click and drag in the main window to select a rectangle that overlay the object or reticule in such a way that the dimensions of the object can be matched to those of the rectan gle fig 4 34 Note that this rectangle is red If the rectangle is incorrectly drawn simply redraw it until you are satisfied with its size and position A calibration only needs to be performed in one dimension usually horizon tal or vertical if the dimensions of the image are isotropic the test object or reticule Frame Grabber Figure 4 34 Calibration rectangle 4 Select image calibration from either the popup menu clic
53. gical studies The FlowMeter is supplied with MyoFlow an add on software programme for MyoView Besides calculating the flow rate MyoFlow enables real time calculations of total flow and current drop volume In addition MyoFlow features a real time flow graph showing flow rate development over time DMI Pressure Myograph System Model 110P amp 111P 2 4 2 5 Connecting Oxygen Supply Connect the oxygen supply pipe on the Pressure Myograph Chamber cover to an adjustable oxygen supply 2 4 2 6 Connecting Suction Device Connect the large pipe on the Pressure Myograph Chamber cover see fig 2 2 p 18 to a vacuum pump via a suction bottle and a vacuum valve as illustrated in fig 2 1 p 17 The internal diameter of the silicone tube used for this connection must be 2mm a 2mm tube is not included in the myograph system User Manual vers 3 2 Chapter 2 27 28 Main Menu E i f Reci gt Chapter 3 The Pressure Myo Interface Chapter 3 is a complete manual for the Pressure Myo Interface version 3 2 The chapter contains a step by step description of how to navigate in the menus and how to use the special features of the Pressure Myograph System like the sequencer program 3 1 Myo Interface Manual Fig 3 1 is an overview of the menus in the Pressure Myo Interface version 3 2 The menus are divided into main menus and submenus In the following sections all menus are described in detail Myo Interface M
54. graph System Model 110P i vsnnnvnnnnrnnnnnrnnnnnnrennnnrennnnrnnnnnnennnnerennnsnenn 18 2 3 Setting up Ned 19 2 3 1 Setting up the complete Pressure Myograph SYSt M ccccscsssssscecescesssereeseccsssereeseceas 19 2 3 2 Installation of Frame Grabber Drivers and MyoView M 1 nnrarnvnnnnnnnnnnnnnnnnnnnnnnnnnnnnennnnnnssennn 20 2 3 2 1 Picolo Frame Grabber Driver Installation rasnrvnranarnnrnannanernnnnnnnnnnnnnnnnnnnnnnernnnnunenrnnnunnennr 21 EMS NN 25 2 4 1 The first Weight and Pressure Calibrations rrnnnnenvnnnnvnnnnnnnnnnnnnvnnnvnnnnnnnnnnnnnnneenennnnnnnnnnnnnn 25 2 4 2 External Pressure Myograph Unit CONNECIONS unable 25 2 4 2 1 Connecting Perfusion eee ken 25 ENE AT NN 26 2 4 2 3 Connecting Superfusion Flow OptiOnall cccccccecssssseeeeeeseesseeeeceeeeaseseeseseeaseseseeseseas 26 2 4 2 4 Connecting Flowmeter Model 161FM Optional ccs sssceeeccessseeeeeeccesseereeesceas 26 2 4 2 5 Connecting Oxygen Supply rrnernnnannnnrnnnnnnnnnnnnnnnnnnnnnnnnnrnnnnnnunenernnnnnnunenernnnnnnenevennnnnnusveesnnn 27 2 4 2 0 Connecting NNN 27 Chapter 3 The Pressure Myo Interface annnannnannnnnnnnnnnnnnnnnnnnnnnnnnvnnnnnnnnnnennnnn 28 3 1 Myo lnterface Manual ansees kdansdaddendd 28 SETTE FUNCIONS Sn 29 31 TUTON EE oct eiin su atapntaee eee eueaeentacat 29 BENN 29 SS SUNN MeN EE NN ENEA EAN 30 SAM CO 0 CU EE EN ME ascent 31 NNN Nr 32 Chapter 4 MyoView Software Manual rnanunannnnnnnnnn
55. h a temperature probe e Power cord the shape of the AC plug varies by country be sure that the plug has the right shape for your location e Serial Cables for PC connection or serial connection to other DMT Pressure Myograph Systems 5 Pressure Regulator e Power cord the shape of the AC plug varies by country be sure that the plug has the right shape for your location e Atlas Copco Cablair hose for main pressure supply e Two 500ml Schott Duran Bottles incl special designed screw caps 6 Riester Big Ben Pressure Manometer User Manual vers 3 2 Unpaking the myograph system 11 12 7 Accessories e Allen key e Spare screws e Small screwdriver e Suture wire 3m e 1 tube of high vacuum grease e 1 tube of grease for linear slides e Silicone tubing internal diameter 1mm 25m 8 Hardware Software amp Manuals e Euresys Picolo Frame Grabber PCI Card e User manual Pressure Myograph System Model 110P amp 111P e MyoView Pressure Myograph software including drivers CD ROM 9 FlowMeter Model 161FM Optional e Euresys Picolo Frame Grabber PCI Card e User manual FlowMeter Model 161FM e MyoFlow FlowMeter software including drivers CD ROM 10 Peristaltic Pump for Superfusion Flow Optional e Alita Watson Marlow400 VS2 10R Midi 2 5 50rpm 11 Computer Optional DMI Pressure Myograph System Model 110P amp 111P Chapter 1 System overview 1 1 Pressure Myograph Unit Horizontal A
56. he left in fig 4 1 show the frame grabber image the analysis image data controls parameter controls and sequencer Each of the control bars can be undocked resized removed or shown in the main window all decided by the user This allows the user to design and configure their per sonal interface set up found to be appropriate for any particular experiment Once the user has designed an ideal interface configuration it may be saved for future applica tions Multiple interface configurations may also be created and saved if more than one user is using the same software At any time users can always return to their default interface configuration or to the factory configuration User Manual vers 3 2 Chapter 4 35 4 2 User Interface 4 2 1 Menus Menus are located across the top of the MyoView interface All the actions available using the control bars are also available from these menus This common windows feature is very useful for example when the control bars are turned off Fig 4 2 Shows how the frame grabber menu is used to save the current image to disc MyoView 1 1 Frame Grabbe Save curent image Fenit fico Li Qulflow Pressure mn Figure 4 2 Access to MyoView functions via the menus 4 2 2 Control Bars Control bars are generic to the MyoView application and give the software a particu lar flexibility which users may find helpful in organising their work Since the Control Bars m
57. hen stable Force 2245 F3 Calibrate Force DONE Push Esc Force Zero 20315 Force gain 668 3 Temp auto 22 4 c pH output 2mv F3 Calibrate pH Slope Slope buffer 4 00 Temp auto 22 4 c pH output 177mV F3 Calibrate pH OFFSET Offset 2208 Slpoe 26 4 3 1 1 General Key Functions Tet Scroll up and down in the menus Being in the top line of a menu use fT to change to the previous main menus Info Push this key in any menu to display all specific F key functions Numeric keys Enter data or setting values Enter Confirm entered numeric values or change between available choices Esc Delete incorrectly entered numeric values In some submenus used to return to main menu F1 Change to the next menu F2 F4 Have different but specific functions dependent of the active main menu 3 1 2 Turn on Message Turning on the Pressure Myo Interface a message appears as shown to the right Use the f and keys to adjust the display Myo Interface Model 110P contrast setting Version 3 2 Press Fi for proceeding to the Main Menu 3 1 3 Main Menu The Main Menu displays P1 Inlet pressure reading in mm Hg P2 Outlet pressure reading in mm Hg Fo ree NE Force Longitudinal force reading in mN 55 3 mm Hg Temp Actual temperature probe reading in C Settemp Heating temperature setting in C Mg Heat Actual heating status ON OFF 7 00 pH Actual pH reading Opt
58. ick Next To install to this folder click Next To install to a different folder click Browse and select another folder Destination Folder C Program Files E uresys Picolo Industrial Browse lt Back Cancel InstallShield Wizard Setup Type choose Typical ke Setup Type to install f Dy Cl i K N ext Click the type of Setup you prefer then click Next Typical Program will be installed with the most common options Recommended for most users Compact Program will be installed with minimum required options C Custom You may choose the options you want to install Recommended for advanced USEIS lt Back Cancel User Manual vers 3 2 Chapter 2 21 InstallShield Wizard Click Next Select Program Folder Please select program folder Setup will add program icons to the Program Folder listed below You may type a new folder name ar select one from the existing folders list Click Next to continue Program Folders Euresys Picolo Industrial Existing Folders Accessories Administrative Tools Games Java Web Start Printhe Internet Printing Startup lt Back Cancel InstallShield Wizard Click Yes Euresys Sample programs Disclaimer Please read the following Disclaimer Press the PAGE DOWN key to see the rest of the text Sample programs Disclaimer You have choosen to install one or more sample programs provided in the Picolo Industrial packag
59. ional Use the f and keys to see all values as the display is only capable of showing four lines at the time Temperature setting To change the temperature setting move the Set Temp line to the top of the display Use the numeric keys to enter a new temperature setting and press Enter to confirm Heating control To turn on heating move the Heat line to the top of the display and use Enter to switch between on and off User Manual vers 3 2 Chapter 3 29 30 The Main Menu has the following functions Info Press the Info key and the display shows the F key op tions in the Main Menu Press Info again to return to the Main Menu or one of the F keys to proceed with one of these options F1 Press FI to proceed to the Pressure Menu F2 Press F2 to start the pressure control programme The settings of the pressure control programme are adjust ed in the Pressure Menu F3 Press F3 to zero the longitudinal force reading F4 Press F4 for the display to show a condensed Main Menu showing pressure inlet reading P1 pressure inlet setting P1 set pressure outlet reading P2 pressure outlet setting P2 set force reading F pH reading Temperature reading T and temperature set ting T set 3 1 4 Pressure Menu The Pressure Menu displays P1 Inlet Inlet pressure setting in mm Hg P2 Outlet Outlet pressure setting in mm Hg Status Activation of pressure con
60. k that the Myo Interface and the Pressure Myograph Unit are properly connected through the 25 pin grey cable 2 Ifthe pressure reading still appears unstable then perform a new pressure cali bration of the pressure transducers as described in Section 5 1 4 3 During the pressure calibration monitor the three relative pressure reading values in the calibration menu on the Myo Interface e If the value is O or above 6500 for one of the pressure transducers then the pressure transducer is broken and needs to be changed e If the reading is between 1 499 or 5000 6500 then contact DMT for further instructions If the message OFF is displayed in either of the pressure reading lines in the Main Menu on the Myo Interface then the pressure transducer is broken and needs to be replaced In case of any other problems related to the pressure transducer please contact DMT for further instruction and advice DMI Pressure Myograph Systems Model 110P amp 111P 5 2 The DMT Microscope Model 111P The DMT Microscope is an inverted microscope equipped with a Zeiss Achromat 10X 0 25 objective and build in CCD camera The objective is adjustable in all X Y Z directions using the three micropositioners on the front as illustrated in fig 5 14 Figure 5 14 How to adjust the focus of the microscope in the X Y Z directions The MyoView data acquisition and analysis software obtains an image by measuring the differences in light inte
61. k on FN on the frame grabber control bar or select the calibrate image icon on the tool bar A dia log box will appear to enter the rectangle dimensions appropriate for the known dimensions of the test object or reticule Note that if the isotropic option is se lected only one dimension is required in this case the x dimension is 328 543 microns to set the other Calibrate rectangle Rectangle dimensions Units 328 a43 Y ji 54 162 R 362 914 W Calibrated Figure 4 35 Calibration of image analysis micron Magnification Calibration File Load Save Filename ftest cal Cancel User Manual vers 3 2 Chapter 4 55 5 Once the dimensions and units of the calibrated rectangle are correctly entered press the calibrate button on the dialog box The MyoView image analysis is now calibrated All parameters passed to other control bars or logged will be cali brated 6 To save the calibration settings for future experiments a calibration file cal can be saved and subsequently loaded from the calibration dialog box The filename being used for calibration is displayed in the dialog box to remind the user of the current settings The user may also change the scaling applied to the current calibration parameters from the calibration dialog box or by using the adjust calibration icon Zill on the tool bar This facility can be useful if MyoView is
62. le the box you once received the myo graph system in Place the wrapped items in the middle of the box and fill out the Surroundings with chips of expanded polystyrene Important Before closing the box make sure that no enclosed items can be shaken around as transport by road or air from time to time can be quite roughly Address the box to DMT A S Skejbyparken 152 DK 8200 Aarhus N Denmark Make sure that all four sides of the box are marked fragile or similar Make an indi cation on the top of the box that it contains goods returned for repair service Customers outside the EC must further enclose a pro forma invoice stating that the box contains goods being returned for repair or service If arranging transportation through a courier please keep in mind the high value of the myograph system and that a standard insurance provided by the courier in most cases is insufficient to cover damage or loss of the myograph system In most cases an additional insurance coverage is needed User Manual vers 3 2 Appendix 4 93 94 Appendix 5 System Specifications Chamber Unit Vessel size Vessel alignment Mounting supports Chamber Chamber material Chamber volume Chamber cover Force range Force resolution Pressure range Pressure regulation Heating Temperature range Temperature resolution Temperature probe Weight calibration Pressure calibration Analog output Digital output Voltage D
63. led Click Finish Click Finish to complete Setup lt Back Data acquisition and analysis software and frame grabber drivers have now been installed and are ready for use with the Pressure Myograph System DMT Pressure Myograph System Model 110P amp 111P 2 4 Experiment Setup 2 4 1 The first Weight and Pressure Calibrations Prior to the shipment of the Pressure Myograph System it has gone through two days of continuous testing including final weight and pressure calibrations However in order to ensure that the myograph is working at highest performance DMT recom mends that new weight and pressure calibrations are performed before starting to use the Pressure Myograph System The weight and pressure calibration procedures are described in detail in Chapter 5 1 4 p 63 2 4 2 External Pressure Myograph Unit Connections This section describes how to connect the perfusion circuit the superfusion circuit oxygen supply pressure supply and suction device Perfusion outlet Suction pipe to vacuum Superfusion outlet Perfusion inlet 0 9 0 ee ee s Extra perfusion Superfusion Pipe for oxygen supply Access hole for Extra perfusion outlet inlet reagents buffer inlet temperature or pH probe Figure 2 3 External Pressure Myograph connection 2 4 2 1 Connecting Perfusion Flow The perfusion flow is controlled by the Pressure Regulator via the Myo I
64. male to the D SUB 15 female connector on the frame grab ber board in the PC Connect the phone plug to the camera s video output and the DC plug to power input 6 Myo Interface External Data Acquisition System Connection optional The four BNC ports on the Myo Interface front enables connection of an external data acquisition system for recording the inlet pressure the outlet pressure the longitudinal force the actual temperature and the pH value of the superfusion or perfusion buffer Note Only four of the five data channels can be recorded simultaneously 7 Turn on the Power Turn the main power to the Myo Interface and the Pressure Regulator on at the power switch also the Zeiss Microscope if you have purchased the Model 110P Turn on the computer and the Pressure Myograph System is now ready to be used with the MyoView software Note A detailed guidance in how to connect Pressure Myograph System for an experimen tal setup is described in detail in Chapter 2 4 p 25 2 3 2 Installation of Frame Grabber Drivers and MyoView This section describes how to install the Eyresys Picolo frame grapper PCI card in your computer along with drivers and MyoView software Note If you have purchased a computer from DMT in conjunction with your Pressure Myo graph System the card drivers and MyoView software have already been installed for you 1 Turn off the computer and install the Eyresys Picolo Frame Grapper
65. n 1 25 using double distilled H O 4 Add 1 091 g l Glucose 2 100 g l NaHCO D Aerate the solution with carbogen 95 0 5 C0 for at least 20 minutes If necessary wait further for the pH of the buffer to reach pH 7 4 User Manual vers 3 2 Chapter 7 87 Appendix 1 Pressure Myograph Set up Preparation Prepare PSS buffer and other solutions Aerate PSS buffer with 5 CO2 and 95 O Rinse myograph and tubing from the pump unit with double distilled water or MQ Fill the myograph and tubing with 37 C PPS Prepare artery without branches and mount it on to the two glass cannulas keep some PPS cold Keep 250ml PSS depending of experiment in water bath 37 C while airing the solution Connection pumps setup myograph Fill the P1 bottle right one from pump unit with 150ml PSS and close the bottle 150ml in case you do not use flow and the artery will just be pressurized other wise you need more Turn the air on the backside of the pump unit ON pressure around 0 5 bar Turn on both interface and pump units backside Put myograph on microscope stage valves closed switch on microscope Connect the computer cable to the myograph and start computer Myoview Turn heat ON on the pump unit interface main menu use F1 to look at different menu scroll within the menu with the arrows Air the myograph chamber and put in the temperature probe On pump unit interface make sure both flow and status are off and set P
66. n 8 acetic acid solution for 10 minutes and continue the mechanical cleaning with a swab stick 6 Wash the myograph chamber several times with double distilled water User Manual vers 3 2 Chapter 5 71 78 Chapter 6 Myograph Accessories and Spare Parts This chapter contains a complete register of equipment needed to set up a basic Pressure Myograph System In addition the chapter contains a list of special pressure myograph accessories and spare parts Besides the main focus on developing and manufacturing of myograph systems DMT has specialised in offering our costumers first class laboratory equipment needed for a basic myograph setup at very competitive prices Contact DMT Sales Department for further product information and prices on any of the products listed below 6 1 General Myograph Equipment This section contains a complete and yet very useful checklist of laboratory equip ment needed when setting up a basic Pressure Myograph System e Dissection Stereo Microscope Incl Ocular micrometer and stage micrometer DMT recommends Zeiss Stemi 2000 Stereo Microscope e Dissection Scissors Item DS 1000 DMT recommends Geuder G 19745 8 cm straight trabeculum scissors e Dissection and Mounting Forceps Item DF 3000 DMT recommends Dumont Medical No 5 tip 0 10mm x 0 06mm e Pipettes DMT recommends CappAero pipettes e Light Source DMT recommends Schott Cold Light Source either Model KL 200 or Model KL 1500
67. n to the right The first line shows the value of the offset buffer first buffer solution which is always 7 00 The second line shows the temperature of the buffer solution The temperature is an important parameter in the calibration formula and is obtained automati cally by placing the myograph temperature probe in the buffer solution The third line shows the output from the pH probe as raw data from the A D converter Place the pH meter electrode and temperature probe in the offset buffer solution and turn on stirring When the relative pH output in the bottom line is stable press F3 to proceed In the first line is now shown the value of the slope buffer second buffer solution which is always 4 00 Place the pH and temperature probes in the slope buffer solution and turn on stirring When the rela tive pH output in the bottom line is stable press F3 to proceed The calibration is now finished The parameters are stored in the internal memory of the Myo Interface Press ESC to return to the Setup Menu User Manual vers 3 2 F4 Calibrate Force Apply no Force Push F3 when stable Force 1938 F3 Calibrate Force Place 2 g on the pan Push F3 when stable Force 2245 F3 Calibrate Force DONE Push Esc Force Zero 20315 Force gain 668 3 F4 Calibrate pH OFFSET Offset buffer 7 00 Temp auto 22 4 c pH output EAA F3 Calibrate pH Slope Slope buffer 4 00 Temp auto 224 C pH
68. nd Down Arrow Keys Scroll up and down in the menus If you are at the top ofa i menu use to change to the previous main menu Analogue Recorder Output Connection to external data acquisition system optional Myo Interface Display e EP 0 o cee oe O F Keys Have different but specific func Numeric Keys Enter data or setting values tions dependent of the active main menu Enter Key Confirm entered numeric values or INFO Key Push this key in any menu change between available choices to display all specific F key functions cesooscoosoocooooooocoooooooooo QD ESC Key Delete incorrectly entered numeric values In some submenus used to return to main menu Figure 1 3 Pressure Myo Interface Front 115 230 V 50 60 Hz Automatic Voltage Selector prsseensseeneceeneneeneneeneseensneensneanssgenssnunsnansonasonnssgnaconasesnasesnssesneseeneseenene e ON OFF Switch prreveesvevarvavasesvasasvavasvavasvanasesvavasvenaseavasesvasasvavasvavasvsvasesvevasvanasssvasenser Power Connector REGU ATOR RS232 12V DC MYOGRAPH Connection Port for con Connection Port for Pres trol of Pressure Regulator sure Myograph Unit b RS 232 Port for Serial Connection to PC Connection Port for pH Meter optional Ter TE e 12V DC Port Figure 1 4 Pressure Myo Interface Back User Manual vers 3 2 Chapter 1 16 1 4 Pressure Regulator Pressure and
69. nect the silicone tube between the outlet connecting pipe and the outlet pressure transducer connecting pipe fig 5 12 A Connect the enclosed resistance tube shunt to the stainless steel pipe inside the myograph chamber and the outlet pressure transducer connection pipe as illustrated in fig 5 13 A B Figure 5 12 Disconnection of silicone tubes User Manual vers 3 2 Chapter 5 67 68 Figure 5 13 Mounting of Resistance tube shunt 6 Close the inlet from the pressure regulator to P1 by turning the handle on the 3 way valve towards the normal inlet way The inlet to PL now comes from the pressure manometer See fig 5 3C p 61 and fig 5 1 p 67 T7 Close the calibration valve on the front of the Pressure Regulator by turning it clockwise Important Make sure that the calibration valve is completely closed The calibration valve not being completely closed results in an inaccurate pressure calibration and unreliable pressure readings and settings during an experiment 8 10 12 Turn on the Pressure Regulator and the Myo Interface Use F to go to the Setup Menu in the Myo Interface Bring the calibrate line to the top of the display by using the numeric keys and press Enter to choose between calibrating pressure force or pH Choose Press in the calibrate line and press F4 to enter the calibration menu Apply a pressure of 50mm Hg on the pres
70. nfo key and the display shows the F key op tions in the Pressure Menu Info Press Info again to return to the Pressure Menu or er one of the F keys to proceed with one of these op tions F1 Press Fi to proceed to the Sequencer Menu F2 Press F2 to start the pressure control programme 3 1 5 Recorder Menu The Pressure Myo Interface has four analogue output ports at the front panel for connection to a data recording system een aT Each of these output ports can be programmed to represent Rec 1lo 0 inlet pressure reading outlet pressure reading longitudinal More force reading temperature or pH meter The default settings ia a n Rec 3 gt Force are Rec 3 lo SO Rec 3 hi 100 Rec 4 gt Temp Rec 4 lo z Rec 4 hi 050 P1 Inlet pressure output P2 Outlet pressure output Longitudinal force output Temperature output The full scale output on the recorder is one volt In the Re corder Menu it is possible to change the value that corre sponds to 0 V lo and 1 V hi User Manual vers 3 2 Chapter 3 31 32 3 1 6 Setup Menu The Setup Menu displays Calibrate Choose which sensor to be calibrated Pressure force or pH Reservoir Temperature setting for perfusion buffer reservoir Option The option line allows access to submenus using Special codes To change any of the settings in the Setup Menu move the line to be changed to the top line of the displa
71. ng together e Turn pump units off and close the air vault at the back of the pump unit e Fill myograph with its tubing and tubing of the pump units with acetic acid 10 let it rest for 5 minutes and rinse everything with MQ 4 times acetic acid must be totally washed out e Rinse P1 bottle and other glass with acetic acid as well e Close program computer DMI Pressure Myograph System Model 110P amp 111P Appendix 2 Terms of Warranty Warranty DMT A S warrants to the original user that myograph systems manufactured by DMT A S will be free from defects in materials and workmanship for a period of three years after the date of delivery DMT A S will repair or replace any defective part subject to the conditions limitations and exclusions Exclusions Force and pressure transducers separately or part of myograph systems manufac tured by DMT A S are disclaimed from any warranty Limitations This warranty shall not apply to equipment subjected to accidental damage improper use alteration or deterioration Warranty on third party products will be as determined by their respective manufac turer DMT A S shall not be liable for consequential incidental special or other direct or indirect damages resulting from economic loss or property damage sustained by you or any end user from the use of the products sold or services rendered hereunder Warranty Returns A Return Material Authorisation RMA number is required f
72. nits are image pixels However if the dimensional analysis has been calibrated as in the example shown the dimensions will be displayed in microns default or in another selectable unit When performing calibrated meas urements the calibration file cal from which the scaling data are taken will be displayed in the dialog box If the calibration has been performed but not saved to file not recommended measurements will be displayed in real units but no filename will appear in the dialog box Measure rectangle calibrated x Calibration File Load ave Units microns eotapit Rectangle dimensions a aso 134 78 Fd oa R 271 L W Calibrated Magnification Filename li ftest cal Figure 4 17 Calibrated distance measurement User Manual vers 3 2 Chapter 4 45 4 3 4 Data The data control bar supervises the acquisition Data FT logging and display of experimental data Some of the data fields are direct readings of experimental s Inflow Pressure mmHg settings e g temperature whilst others are de p Cutfiow Pressure mm rived from selected analysis within a given experi te Mean pressure mmHg ment e g lumen diameter These data fields may Target Inflow Pressure Target Outflow Pressur Temperature deg C be displayed graphically and or logged to file as s required The graphical display is always present
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74. nsity passing through the walls of a vessel segment mount ed in the Pressure Myograph Chamber Traditionally a white light source is sufficient for such a purpose but has one major disadvantage A white light source makes the data acquisition and analysis very sensitive to changes in white light intensity from the surroundings such as room and sunlight To avoid the surroundings from having influence on the data acquisition and analysis the DMT Microscope has a build in infrared light source 5 3 Zeiss Axiovert 40 Microscope Model 110P Pressure Myograph System set ups require high flexibility in image analysis and DMT recommends the Zeiss Axiovert 40 inverted microscope The advantage of the Zeiss Axiovert 40 is the high degree of flexibility and upgrades available This both include a wide range of different objectives equipment for sophisticated contrast and fluores cence techniques and various documentation features Contact DMT Sales Department for further product information and prices on any optional equipment for the Zeiss Axiovert 40 Product information is also available at WWW ZeISS COM 5 4 The Pressure Regulator The Pressure Regulator controls the flow and pressure applied to the vessel mounted User Manual vers 3 2 Chapter 5 71 72 in the pressure myograph chamber via settings in the Myo Interface and the Myo View software This Chapter is intended as an overview of the Pressure Regulator to understand how it works
75. nterface and the MyoView software How to control the perfusion flow is described in detail in Chapter 3 1 4 p 30 Chapter 4 3 5 p 48 and Chapter 4 4 p 54 To connect the Pressure Regulator to the Pressure Myograph Unit use the silicone tube internal diameter 1mm delivered with the Pressure Myograph System Connect the perfusion inlet at P1 on the Pressure Myograph Unit see fig 2 1 p 17 and fig 2 2 p 18 to the P1 outlet on the cap of the buffer bottle see fig 2 1 and fig 5 12 p 67 Connect the perfusion outlet at P2 on the Pressure Myograph Unit see fig 2 1 and fig 2 3 to the P2 inlet in the Pressure Regulator Unit see fig 2 1 and fig 2 2 User Manual vers 3 2 Chapter 2 25 26 Note I To fill the tubing and avoid air bubbles mount the Resistance tube shunt as de scribed in Chapter 5 1 4 3 p 67 Fill the Schott bottle with 250 ml buffer and run the following flush programme P1 150mm Hg P2 70mm Hg Enter the pressure values in the Pressure Menu and press F2 in the Main Menu to start the flush programme Run the programme until no air bubbles are visible in the perfusion circuit Note II Never fill the buffer bottle with more than 250 ml of buffer as higher volumes makes precise control of the pressure more difficult Important Never run the flush programme with a vessel mounted in the Pressure Myograph Chamber as this may cause serious damage to the ves
76. nu click on h p on the data control bar When data logging is active the cross indicator becomes a tick indicator L Data logging can be stopped at any time by selecting the stop data logging button Li in the data control bar or by selecting the start stop logging data menu item in the popup menu Note that data will be stored in the data history graph regardless of whether data is logged to disk User Manual vers 3 2 Chapter 4 47 48 4 3 5 Sequencer The sequencer can be used to control the active operating conditions of the Pres sure Myograph System such as inflow pressure outlet pressure temperature and the duration of these specific settings The feature enables the user to programme an experimental protocol consisting of a sequence of specific pressure and temperature settings MyoView enables the user to store programmed sequencer protocols for later replication of a specific experiment Also it is possible to edit a stored sequencer protocol and then store it as a new sequence protocol The operating settings pressure and temperature of the Pressure Myograph System cannot be changed instantly and the time needed to reach a specific setting depends on the current setting The sequencer takes these factors into consideration in the way it controls the experimental design The popup menu on the control bar select xl can be used to access all functions of the sequencer This popup menu can also be r
77. o Reduction Valve Manometer 0 5 Bar Outlet from P2 on Pressure Myograph Unit Relief Valve 0 8 Bar W Air Out pi N m st n E Q Q Q Q au 2 gt s s s s Needle Valve Calibration Valve Figure 5 15 Pressure Regulator Diagram Buffer Flow Air Flow User Manual vers 3 2 Chapter 5 73 5 4 3 Pressure Transducer Calibration Calibration of the three pressure transducers requires that the calibration valve is completely closed The importance of closing the calibration valve is illustrated in the diagram in fig 5 15 p 73 If the calibration valve is not closed then the pres sures applied using the pressure manometer will have access to a huge air reservoir in the left buffer bottle The air reservoir will appear very compressible making the applied pressures and the whole pressure calibration very unreliable The diagram in fig 5 15 p 73 also shows the importance of opening the calibration valve after a pressure calibration If the calibration valve is closed before running an experiment pressure will build up and damage the mounted vessel because the buffer is unable to return to the left buffer bottle 5 5 Pressure Myograph Maintenance and Cleaning The Pressure Myograph System Model 111P amp 110P is a very delicate and so phisticated piece of research equipment In order to keep it working at its best DMT recommend that the following sections are
78. o aj O O Ef np hH E Ey a et i mig i a i ell m Mu mm Te mo D irimi i Dow ow a pila ne u Mn w AX wm Ao Cone cine D Ow OM w Figure 4 37 3wa m A 3 3 bien D 1 X Custome inter r wal oon an x1 z face set up A DMT Pressure Myograph System Model 110P amp 111P 2 Design of a custom interface is done through reconfiguration the control bars and main window contents In the example shown in fig 4 37 the parameter data and sequencer control bars have all been removed The easiest way to do this is from the tilj a Slicons on the tool bar The graphical contents of the analysis con trol bar have been selected to the main window and subsequently the analysis control bar itself has been removed using the Toolbar icon R Finally the frame grabber control bar has been undocked resized and positioned in the centre of the main window In this user setup the image processing output may be monitor ing closely whilst maintaining observation of the whole image in real time kue 11 Seen semje al reel mm ep 154 macros HH H 120 microns aa Figure 4 36 MyoView main window with factory settings 3 To save a setup for future use select save from the user setup options and se lect an identifier number from the options This setup can be recalled by select ing load from the user setup menu options User set ups a
79. odel 110P Version 32 F1 Recorder Menu Press 1 gt Setup Menu Calibrate Press Resovoir 2 B G 0 P1 Inlet 64 4 mm Hg P2 Outlet 0 7 55 5 min hes Sias Rec 1 hi 2 200 LOS mi Flow REC 2 gt Press 2 2 2857 atm lever Rec 2 lo 0 Rec 2 hi 200 Rec 3 Rec 3 lo RO Rec 3 hi 2 OG Rec 4 Rec 4 lo 0 Rec 4 hi 50 Rec 1 lo Info Change menu Cal pressure Info Change menu Start pres Ctrl Change menu Start pres Ctrl Zero force Condensed menu F4 Near 12 5mN Far 2 7 mN Act 26 5 Heat OFF Setup Set 37 0 Calibrate Menus Ane LOO Setup F4 F4 F4 Options Menus Calibrate Pressure Calibrate Force Calibrate pH OFFSET 7 r Apply 50 mm Hg Apply no Force Offset buffer 7 00 No 7 Comm Menu Serial mode RS485 Myograph 2 A No 10 pH Menu Offset buffer 7 00 Slope buffer 4 00 Temp comp Auto Manual temp 37 C Push F3 when stable P1 3614 P2 3533 F3 Calibrate Pressure Apply 125 mm Hg Push F3 when stable P1 4252 P2 4222 F3 Calibrate Pressure Apply 200 mm Hg Push F3 when stable P1 4956 P2 4933 F3 Calibrate Pressure DONE Push Esc 36148 42527 49565 35334 42223 49332 Figure 3 1 Menus in the 110 P 111P Myo Interface version 3 2 DMI Pressure Myograph System Model 110P amp 111P Push F3 when stable Force 1938 F3 Calibrate Force Place 2 g on the pan Push F3 w
80. of the Myo Interface Press ESC to return to the Setup Menu DMI Pressure Myograph System Model 110P amp 111P Setup Menu Calibrate Press Resovoir 2 SUG Option 2 Info Change menu Cal pressure F4 Calibrate Pressure Apply 50 mm Hg Push F3 when stable Pi 3614 1P2 3535 F3 Calibrate Pressure Apply 125 mm Hg Push F3 when stable P1 4252 P2 4222 F3 Calibrate Pressure Apply 200 mm Hg Push F3 when stable P1 4956 P2 4933 F3 Calibrate Pressure DONE Push Esc 36148 42527 49565 35334 42223 49332 For a more detailed instruction to the calibration procedure see Chapter 5 1 4 p 63 Force transducer calibration Chose Force in the calibrate line and press F4 to enter the window shown to the right Make sure that no force is applied on the force trans ducer and when the relative reading at the bottom line of the window appears stable press F3 to proceed Carefully place the 2 gram weight on the load pan and when the relative reading at the bottom line of the window appears stable press F3 to proceed The calibration is now finished The readings in the two bottom lines are the calibration parameters stored in the internal memory of the Myo Interface Press ESC to return to the Setup Menu For a more detailed instruction to the calibration procedure see Chapter 5 1 4 p 63 pH calibration Optional Choose pH in the calibrate line and press F4 to enter the window show
81. oosening of screw F fig 5 2 Important Be very careful not to apply too much force gt 100 gram on the force transducer pin when adjusting the left glass cannula The force transducer has a mechanical protec tion in its longitudinal direction of movement but is very vulnerable to force applied on the force transducer pin Right Glass Cannula Horizontal positioning of the right glass cannula is adjusted by carefully loosening screw A fig 5 2 Vertical positioning of the right glass cannula is adjusted with the micropositioner C fig 5 2 Longitudinal positioning of the right glass cannula is ad justed by gently loosening of screw G fig 5 2 Note Be careful not to damage the tip of the glass cannulas during the general adjustment 5 1 2 3 Way Valve Adjustments The 3 way valves on each side of the Pressure Myograph Unit at P1 amp P2 have three different settings to control the in or outlet flow to the pressure transducers The flow settings are illustrated in fig 5 3 and fig 5 4 Figure 5 3 Three way valve flow settings at P1 Figure 5 4 Three way valve flow settings at P2 User Manual vers 3 2 Chapter 5 61 62 5 1 3 How to Read a Millimetre Micrometer Sleeve Scale Timble Scale R E S 4 gt Figure 5 5 Overview of a Micrometer Actual reading 20 00mm Sleeve Scale The micrometer sleeve scale has a total length of 25mm divided into fifty equal parts Each part of division above the h
82. or all returns This number should be clearly indicated on all returned myograph systems Products damaged due to improper or inadequate packaging when returned for RMA purposes are not granted warranty coverage User Manual vers 3 2 Appendix 2 91 92 Appendix 3 Service Check A myograph working at optimal performance is extremely important for Success when studying small blood vessels or other small tubular tissues To make sure that our customers always are dealing with first class myographs DMT offers a Myograph Service Check at a very favourable price The Myograph Service Check includes a complete separation for inspection of all mechanical and electronic parts The myograph is then reassembled adjusted and finally all electronic and mechanical parts are tested Please note that the service does not include replacement of transducers or any other needed spare parts Please contact DMT for information about prices DMI Pressure Myograph System Model 110P amp 111P Appendix 4 Shipping Instructions If the myograph system needs to be sending back for service or repair please read the following shipping instructions very carefully Before you start packing the myograph system please remember that you are dealing with very delicate equipment and therefore care must be taken DMT recommends that each part of the myograph system be wrapped individually i e with bubble wrap and placed together in a large box preferab
83. orizontal line represents 1mm where each fifth line is marked by a longer line and a number which designates the length in mm Each part of division below the horizontal line is placed between each 1 mm mark scale above the horizontal line and represents 0 5mm Thimble Scale The thimble is divided into fifty equal parts and one complete rotation of the thimble indicates with the smallest division on the sleeve equalling 0 5mm Reading Example 1 1 Note that the thimble has stopped at a point be yond 10 on the sleeve indicating 10 00 dd 2 Note that there is no mark completely visible be tween the 10 mm mark and the thimble 3 Read the value on the thimble corresponding to the Figure 5 6 Example 1 intersection with the horizontal line on the sleeve Reading 10 38mm A Reading on sleeve 10 00 B No additional mark visible 0 00 C Thimble reading 0 38 Total Reading 10 38 DMT Pressure Myograph System Model 110P amp 111P Reading Example 2 1 Note that the thimble has stopped at a point beyond 16 on the sleeve indicating 16 00 2 Note that this time a mark is visible between the 16 mm mark and the thimble indication 0 50 Figure 5 7 Example 2 3 Read the value on the thimble corresponding to the Reading 16 78mm intersection with the horizontal line on the sleeve A Reading on sleeve 16 00 B One additional mark visible 0 50 C Thimble reading 0 28 Total Reading 16 78 5 1 4
84. output e I TNV F3 Calibrate pH OFFSET Offset 2208 Slpoe 1o DOM Chapter 3 33 Special Communication submenu To open the special Communication submenu enter the code 7 in the option line in the Setup Menu Serial mode RS232 or RS485 serial communication No 7 Comm Menu setting Serial mode RS485 Myograph A Myograph Myograph serial identification A B Cor D In the Serial mode the serial communication can be chosen to be either RS232 or RS485 RS485 is used when more than one myograph is connected to the serial port on the same computer Up to four myographs can be connected to the same computer but the connection requires a separate RS232 RS485 converter The Myograph line is only shown if the RS485 setting is chosen in the Serial mode line The Myograph line is used to select the number for the individual myographs A B C or D Press F1 to return to the Setup Menu Special pH submenu To open the special pH submenu enter the code 10 in the option line in the Setup Menu Offset buffer pH value of offset buffer Slope buffer pH value of slope buffer No 7 pH Menu Offset buffer Temp comp AUTO or MANUAL temperature compensation Slope bite Manual temp Manual temperature setting Temp comp Manual temp The pH submenu contains the settings of the pH calibration procedure Offset buffer and slope buffer are the pH values
85. oxic chemicals incubate the myograph chamber and glass cannulas with 1M HCI for up to 1 hour In exceptional cases incubate the chamber and supports with a up to 3M HNO3 solution for about 15 minutes 5 Wash the myograph chamber and glass cannulas several times with double dis tilled water Important Notes e Be very careful using step 3 and 4 repeatedly times as strong reagents can cause extreme damage to the myograph unit e Be very careful not to damage the glass cannulas during the cleaning procedure e After cleaning ALWAYS check that the greasing around the transducer pin is suf ficient to keep out the buffer solution from the transducer compartment In cases of red or brown discolorations appearing on the chamber sides the following cleaning procedure will work in most cases 1 Incubate the myograph chamber for 30 minutes with 20ul of a 2mM T 1210 Tetrakis 2 pyridylmethyl ethylenediamine solution dissolved in double distilled water 2 Use a swab stick to mechanically clean all the affected surfaces during the last 15 minutes of the incubation period 3 Wash the myograph chamber several times with double distilled water 4 Incubate the myograph chamber with 96 ethanol for 10 minutes while continu ing the mechanical cleaning with a swab stick 5 Remove the ethanol solution and wash a few times with double distilled water 76 DMI Pressure Myograph System Model 110P amp 111P Incubate the myograph chamber with a
86. puter aided experimental system The designers have tried to give the myograph users a totally flexible design palette from which they can create experi ments with precise control We would be very pleased to receive suggestions as to how the software could be improved and indeed any feedback is always most wel come Please contact the software support team with your comments User Manual vers 3 2 Chapter 4 41 42 4 3 Topics 4 3 1 View The view menu may be used to control the appearance of control bars in the inter face Some user configurations will require certain control bars to be invisible al though it should be noted that it does not affect the function of the control bars For example video frames may be acquired and processed without necessarily displaying the frames either in the frame grabber control bar or main window Fig 4 13 shows as an example how to turn off the analysis control bar The control bars can also be turned on and off by selecting the appropriate icons in the toolbar All control bars can be undocked and re sized for convenience in a given experi ment Once re sized a control bar may be docked but careful sizing and positioning is required if the User Interface is to maintain a well balanced appearance Users are advised to experiment with these facilities to fully appreciate their potential Pome Frame Gabler Dota Sequencer Anayet Help foveGobbe ajajajaja rjeje kj ra elr Figure
87. r is active or otherwise User Manual vers 3 2 Chapter 4 53 54 4 4 How to Implement an Experimental Protocol The MyoView software may be used to record and run an experimental protocol by use of the sequencer This involves both custom settings of all the myograph features as well as the analysis features of MyoView These protocols may be saved to en able precise reproduction of the experimental method The data handling features of MyoView allow experimental recording to be logged automatically so that extended experimental procedures may be conducted with the minimum of operator interven tions A protocol may be designed and implemented by following the procedure Write down the steps of your protocol as a sequence in time thinking about when the myograph control settings need to change Remember that certain changes can require a relative long time to reach their target values e g temperature changes therefore your protocol should include indication of when the system needs to wait until When you are satisfied with the timing of events in your protocol programme the events into the MyoView Sequencer Sequencer f fe fa Ks 0 Stat T p Edit entry a Remove entry Cur 2 Clear all entries z Load sequence Save sequence Start sequencer Start at Pauze sequence Continuous loop Figure 4 31 Programming the MyoView sequencer Launch MyoView and make sure the sequencer control
88. re stored even when the computer is turned off and they will remain unchanged until reset from the User Setup menu options kn i r 154 microns 120 microns Figure 4 38 How to save a customer inter face setup User Manual vers 3 2 Chapter 4 57 58 4 7 FAQ There are only six parameters in the graphical display how do I display other data acquired or calculated by MyoView All the data acquired or calculated within MyoView can be displayed but the maxi mum number appearing in any given display is six The data control bar is used for selecting the appearance of the display Regardless of what the status of the data display is all parameters are calculated and stored internally This means that the history of each parameter may be viewed at any time by recalling that parameter to the display Note that data logging is a separate function within MyoView and should be distinguished from data display Some parameters are always available to be selected for graphical display in the data control bar and others are not Why The data control bar only allows parameters to be selected for display if they are logically available For example current pressure is always available whereas lumen diameter would not be available if the image analysis was not active I have logged some data to file and have viewed the data using a spreadsheet application Why are there several other columns in the file with no entries
89. read carefully and that the instructions are followed at all times 5 5 1 Pressure Myograph Chamber Pipes To prevent the pipes from being blocked by buffer salt deposits after an experiment use the chamber cover to remove the cleaning solutions Afterwards remove the cover from the myograph chamber and turn on the vacuum pump and vacuum valve for about 10 seconds Wait to turn off the oxygen supply until turning off the vacuum pump Wipe off any buffer remaining on the outside of the pipes using a piece of paper 5 5 2 Force Transducer The force transducer is one of the most delicate and fragile components of the Pres sure Myograph System Therefore careful handling of the force transducer is of most importance to prevent it being damaged The left glass cannula in the pressure myograph is connected to the transducer pin coming through a small hole in the transducer house located on top of the pressure myograph unit as illustrated in fig 5 16 To prevent the buffer from running into the transducer house the hole is filled with high vacuum grease Figure 5 16 Transducer pin hole sealed up with high vacuum grease DMI Pressure Myograph System Model 110P amp 111P As a part of daily maintenance it is very important to inspect the greasing of the trans ducer hole before starting any experiment Insufficient greasing causes damage and malfunction of the force transducer Important 1 DMT recommends that the high vacuum grease se
90. rm of a vessel profile showing the posi tions of the computed edges DMI Pressure Myograph System Model 110P amp 111P 4 2 5 Modality Many aspects of the MyoView software layout are similar to standard Windows ap plications e g Microsoft Office Main menus are available for all actions and dia logue boxes A dockable Toolbar containing icons is also provided for many of the important actions Tool tips are implemented throughout the software reminding users of the function of active controls icons list boxes radio buttons etc when the mouse icon is allowed to dwell over each control In addition to these standard features the MyoView interface has been designed to meet the special display and analysis capabilities of the Pressure Myograph systems Model 111P and 110P These capabilities are encapsulated in five dockable control bars and a main window that may be configured to suit the user The control bars pro vide convenient user access to control analysis display and data logging functions A little investment in learning the functionality of these control bars will be rewarded by the ability to implement sophisticated user designed protocols in an automatic mo dality Once designed these protocols can become standards to be repeated exactly with the minimum of operator intervention MyoView is more than just an interface for the Pressure Myograph System it is a complete com
91. rom the con trol bar The same action is also available by selecting the Start stop image display icon i on the Toolbar The rate at which frames are displayed in the interface de pends upon the size of the control bar and whether frames are also being displayed in the main window 4 3 3 2 Image Calibration The image dimensional analysis can be calibrated in sensible units e g microns but the default calibration is performed in units defined by the image data resolution i e pixels The image calibration procedure is described in section 4 3 6 2 p 51 4 3 3 3 Zooming The zoom feature can be used to focus on a region of interest Zooming does not af fect the image analysis but may be useful in determining a suitable region for analy sis Zooming may be performed before or during analysis without affecting any other feature that may be active Before selecting a region to zoom in on the image should be displayed in the main window Drag and click the mouse to select a rectangle in the main window and then select the zoom in icon amp Jjon the tool Bar Selecting the full size icon may restore the full image 8 DMI Pressure Myograph System Model 110P amp 111P 4 3 3 4 Saving an Image Images can be saved to disc in a standard format bmp for future reference or incorporation into documents Images can be saved even when being acquired and displayed in real time which allows the user anytime to catch an intere
92. s of Weight Calibration revvvornnnnnnnnnnrnnnnnrnrnnnnnnnnnnnnnnnnnnevnnnnnnnnnnnnnennnnnnennnnnnnnnnunnne 63 31A Wole Caprag POC CO UNG svares pa E 65 5 1 4 3 Pressure Calibration Procedure iv cxiiscncvencneenexseexnsennnncetmeceseacevecdavenseiecuetendsvenseuenersenteusteneeerens 67 5 1 5 Replacement of Glass Canis rv 69 5 1 6 Checking the Force ANSA sne eniai 69 SG Simple Force Iransd cer Chet een 69 5 1 7 Checking the Pressure Transducers sicrccvsesnsnsiectxacnerccstaeseeeanenesssveneeiaddeveavevencsuecendenterseeensersst 69 5 1 7 1 Simple Pressure Transducer Check sisi tsicssesisacosvaunacdswieiousacabebeesvauneddaawnatestanniceonwainatenwwandoes TO 5 2 The DMT Microscope Model 111P uvanunnnnannanannnnunnnnunnnnnnnnnnnnnnnnnnannnnnnnnnnnnnnnnnennnunnnne 11 5 3 Zeiss Axiovert 40 Microscope Model 110P runaurranavnnnnnnnnannnnnnnnannnnnannnnnnnnnnnennnuennne T1 oA he Pressure Regulators E 71 5 4 1 Control of Flow and Pressure SCUING Ss icccscnnccssssnasasntacaeucesusextusstastexenesttseiaastabantendossutetanvexsine 12 ANN HONG apes pcg cctcisnet se eaetape E E 2 5 4 3 Pressure Transducer CAlDPQtlONn wwisccsmsdncancacsineeriniwinestednentnnsesaluvsdeinadaantleobadsensiennweedasianiienileis 14 5 5 Pressure Myograph Maintenance and Cleaning cccccsscceccessseeeceesseeeeseseeeeeeaaees 14 5 5 1 Pressure Myograph Chamber Pipes Lveeeamarmmsmnmemmmmgevuenkdeni nn 14 5 5 2 Force 150 assisirenrerr renne narena AEAEE EAEE EEE NENANA EEKE 14
93. s of the Enable disable button Ci or from the popup menu All of the data appearing on this control bar may be displayed and or logged to file by use of the data control bar A more detailed description of the functions available in the control bar follows under these sub headings e Hardware Control e Manual Control of Pressure and Temperature e Automatic Control of Pressure and Temperature e Configure Computer Communications 4 3 2 1 Hardware Control Hardware control refers to the manual or automatic control of myograph pressures and joystick and temperature from the MyoView interface To ensure that the hardware control is enabled select the Enable disable hardware control button on the parameter control bar or select from the popup menu Once hardware control is enabled the pressure and temperature control can be independently disabled from the popup menu Disabling hardware control suspends control of temperature and pressures 4 3 2 2 Manual Control of Pressure and Temperature The edit boxes on the parameter control bar enables the user to change the cur rent temperature and pressures settings When editing is completed in each box hit return on the keyboard to activate the value Indicators are visible on the parameter control bar showing whether a given parameter has reached the target value When the target value has been reached the cross indicator becomes a tick indica tor The target v
94. sel 2 4 2 2 Extra Perfusion In Outlet The extra perfusion in and outlet on the Pressure Myograph Unit makes it possible to add special agents or reagents to the perfusion circuit during an experiment The small handle on top of the valves controls the flow directions through the 3 way valves at P1 and P2 as described in Section 5 1 2 p 61 2 4 2 3 Connecting Superfusion Flow Optional Creating a superfusion flow in the Pressure Myograph Chamber requires an exter nal peristaltic pump which is not part of the standard Pressure Myograph System Contact DMT for further information about recommended perfusion pump models Specifications and prices The superfusion circuit is connected to the Pressure Myograph Unit as illustrated in fig 2 1 p 17 and fig 2 2 p 18 2 4 2 4 Connecting Flowmeter Model 161FM Optional The DMT Flowmeter Model 161FM is optional for flow measurements in the range of 1 1500ul min The Flowmeter is easily combined with any DMT Pressure Myo graph System as illustrated in fig 2 1 p 17 Use of the FlowMeter makes it possible to control intravascular pressure pressure gradient and flow conditions with high accuracy which enables the performance of in vitro studies on vessel segments under conditions very close to those found in vivo Application of the FlowMeter also enables measurement of flow rate as a parameter due to changes in vessel structure and function in various physiological and pharma colo
95. sting event Sometimes such an image may be required during the setup phase of an experiment In this instance it is recommended to stop the frame grabber when a suitable image is acquired To save this freeze frame to file click on the save image icon l in the tool bar A dialogue box will appear to allow the user to define a file name and location for the saved image A file extension bmp will be added automatically to en able other applications office and drawing packages to recognise the file format 4 3 3 5 Frame Grabber Configuration Attributes gain offset saturation of the frame grabber can be controlled from Myo View via the frame grabber configuration dialogue box Access to these controls is available by clicking on the configure frame grabber icon Hifin the tool bar Please refer to the PICOLO technical data for an explanation of these controls Images ob tained under normal conditions will not require adjustment of these controls and the majority of users will achieve optimum performance from the default settings 4 3 3 6 Distance Measurement If the user wishes to measure a distance on the image MyoView can supply the dimensions of any rectangle drawn in the main window Drag and click the mouse to select a rectangle in the main window and then select the measure rectangle icon on the tool bar A dialogue box will appear giving the rectangle dimensions in selecta ble units The default u
96. sure myograph using the Big Ben pressure manometer When the relative readings at the bottom line of the window appears stable press F3 to proceed Apply a pressure of 125mm Hg on the pressure myograph using the Big Ben pressure manometer When the relative readings at the bottom line of the window appears stable press F3 to proceed Apply a pressure of 200mm Hg on the pressure myograph using the Big Ben pressure manometer When the relative readings at the bottom line of the window appears stable press F3 to proceed The calibration is now finished The readings in the two bot tom lines are the calibration parameters stored in the internal memory of the Myo Interface Press ESC to return to the Setup Menu DMT Pressure Myograph System Model 110P amp 111P F4 Calibrate Pressure Apply 50 mm Hg Push F3 when stable Pi 3614 P2 3553 F3 Calibrate Pressure Apply 125 mm Hg Push F3 when stable P14252 P2 4222 F3 Calibrate Pressure Apply 200 mm Hg Push F3 when stable P1 4956 P2 4933 F3 Calibrate Pressure DONE Push Esc 36148 42527 49565 35334 42223 49332 13 Carefully remove the resistance tube shunt and reconnect the tubing to the glass cannulas Open the calibration valve on the Pressure Regulator by turn ing it counter clockwise Important Make sure that the calibration valve is completely opened after the pressure cali bration A half closed calibration
97. the Prompt box so that the operator is reminded of what the interven tion should be Having completed the editing of the entry click OK to record the new entry settings Figure 4 21 Sequencer set tings entry Edit Entry Edit Item 4 EZ pa Operator intervention Annotation Duration 0 Walt Inflow Pressure 40 mmHg F Outflow Pressure 20 mmHg T Temperature 20 deg FP Wait for user f Bromet Adjust concentration of perfusate User Manual vers 3 2 Chapter 4 50 4 3 5 3 Executing a Sequence A sequence loaded into MyoView may be executed from the first entry by clicking the Start button gt on the sequencer control bar The sequence may be started at a given entry by clicking the Start at button gt Both of these actions are also avail able from the pop up menu click on xl on the control bar and from the main menu A sequence may be repeated in a loop by selecting the continuous loop item from the pop up menu click on xl on the control bar 4 3 5 4 Saving a Sequence Sequences created or edited by MyoView can be saved as new sequences by select ing the Save sequence menu item from the pop up list on the control bar or main menu This action will cause a dialogue box to appear to enable the sequence to be named and saved or over written in a standard way 4 3 5 5 Exporting a Sequence as a Text File Sequences created or edited by MyoView can
98. tion subsequent graphical display is paused By selecting the resume button Blithe display will reset to the current time i e time 0 During data scrolling real time data is still passed to the internal memory so that it may be reviewed subsequently Note that the internal memory is only used for graphical display If the user wishes to log data for future reference or export to spreadsheet then the data logging facility must be activated The displayed data history may be reset at any time using the clear data history item on the popup menu Note that this command does not affect data that has been logged to disk 4 3 4 4 Data Logging Data files myo can be created from the data control bar which contains a log of all the controlled or derived parameters The logged files contain all possible parameters in a standard spreadsheet format where each column corresponds to a parameter If a given parameter is not calculated or used it will still appear in the spreadsheet This feature protects the user from forgetting to log a parameter that may be required The log files are text based and may be read and displayed by MyoView or any standard spreadsheet application e g EXCEL Data logging is not automatic and must be activated by selecting or creating a file Files are chosen by clicking the start data logging button gt in the data control bar or by selecting the start logging data menu item in the popup me
99. tocols and procedures describing how to quick ly get started using the Pressure Myograph System The chapter mostly addresses users who are not yet familiar with the pressure myograph technique However it may contain some hints that will also be useful for the experienced myograph user 7 1 Dissection protocol for small mesenteric arteries The pressure myograph technique is versatile in that a large variety of physiologi cal and pharmacological studies of ring preparations from different species can be performed For mostly the pressure myograph is used for investigation of small blood vessels and as an example this Chapter describes the dissection of rat mesenteric arteries 1 A laboratory rat is euthanized in accordance to the local national law and regula tions A midline laparotomy is performed to expose the mesenteric bed 2 Use scissors to remove about 10cm of intestine together with its feeding vascu lature including part of the superior mesenteric artery Be careful not to damage the vasculature during this procedure The proximal end of the intestine section Shall be about 10cm from pylorus Make a cut in the proximal end of the intestine for later identification 3 Place the excised intestine section in a Petri dish about 9cm in diameter coated with a 5mm thick layer of Sylgard at the bottom to hold the fixing pins Immedi ately fill the Petri dish with cold PSS well aerated carbogen See section 7 3 p 86 The dissection
100. ton Make a new square in the main window and click the arrow play on the right window on the bottom diameter will be recorded now Save the file by clicking the arrow on the left adjust graph time to 3600 change 77 h the graph option to lumen diameter vessel diameter and temperature Measure manually by drawing a square between the vessel walls inner diameter and push the button Watch temperature and pressure during the entire experiment green is OK Running experiments When the artery gets pressurized to 0mmHg measure the internal diameter manually this is the initial diameter Change PSS every 15 minutes with pre heated PSS 37 c Remove PSS by suck ing it out of the bath open water tap behind pump unit add new PSS immedi ately Wait until myogenic tone occurs the diameter will reduce to 30 from its initial diameter Check the reactivity of the artery by applying 60mM KCL it will constrict maxi mally and by applying 146mM KCL it will dilate for small arteries User Manual vers 3 2 Appendix 1 89 e Wash in between and after and wait for myogenic tone stabilizes before starting an experiment e Make sure there is 10ml PSS in the myograph add drugs directly to the bath Measure dilation and constriction manually file serves as back up End e Turn heat and gas off reduce pressure to S mmHg and turn status off e Disconnect myograph from computer port and tubing and put tubi
101. trol ON OFF Flow Activation of flow control ON OFF mm Hg sec Setting of pressure changing gradient The Pressure Menu enables control and adjustment of pressures and flow P1 Inlet and P2 Outlet settings control the inlet and outlet pressure of the perfusion buffer The pressure control is acti vated or deactivated in the Status line by choosing ON OFF DMI Pressure Myograph System Model 110P amp 111P Info Change menu Start pres Ctrl Zero force Condensed menu F4 Pils LAL P1 set 140 P2 129 P2 set 130 FIOL oo 265 P207 Tm set 3r Pressure Menu P1 Inlet 90mm Hg P2 Outlet 80 mm Hg Status ROER Flow ON mm Hg sec 10 Important Please note that the P1 inlet pressure cannot be smaller than the P2 outlet pressure If a P1 value is entered which is smaller than the P2 value then the P1 value will automatical ly be set equal to the P2 value Also if a P2 value is entered which is larger than the P1 value then the P2 value will auto matically be set equal to the P1 value A setting where P1 P2 activates the No Flow function in the Pressure Regulator as described in Chapter 5 4 2 The function works to prevent backflow The perfusion flow can be activated or deactivated in the Flow line ON OFF In the mm Hg sec line it is possible to adjust and control the pressure changing gradient The Pressure Menu has the following functions Info Press the I
102. urements of the vessel dimen sions a procedure must be followed as explained in Section 4 5 How To Calibrate the Image Analysis To commence image analysis select the stop start analysis icon l on the tool bar or select the Start image analysis menu item from the popup menu click on zi on the analysis control bar User Manual vers 3 2 Chapter 4 51 52 4 3 6 4 Configuration of Vessel Detection and Analysis VDA The dimensional analysis of vessel images is automatic and contains an algorithm that effectively checks the correct positioning of the vessel and its analysis window If the vessel image within the analysis rectangle clearly Shows the vessel walls with good contrast the algorithm will have no trouble in finding the edges and monitoring their changes throughout an experimental protocol This condition will be clear from the analysis results window showing a clearly delineated edge profile see fig 4 24 and fig 4 25 Frame Grabber Image Analysis Figure 4 24 Analysis rectangle Figure 4 25 Vessel wall analysis If the analysis rectangle does not contain a valid vessel MyoView will indicate that no edge detection is possible see fig 4 26 and fig 4 27 This condition will be clear from the analysis results window showing the cross indicator B Frame Grabber Image Analysis Figure 4 26 Invalid vessel Figure 4 27 No image analysis pos sible If the analysis rectangle contains a v
103. used to review the param eter history at any time but should not be confused with the logging of parameters to file which is a separate feature of MyoView DMI Pressure Myograph System Model 110P amp 111P 4 3 4 2 Configuration of Data Display The display of parameters in Myo View can be configured in the Con ok F figure graph window selected from the Time Range s 60 Data Rate 3 2 Maxima popup menu on the data control bar In the configure graph window shown in fig 4 19 it is possible to configure the axis range of each displayed param eter the data rate and the time range Once an appropriate graphical display setup has been entered click the OK bottom to complete the selection Poms neha Cancel T Legend Inflow Pressure Outflow Mean pressure Target Inflow Target Outflow Figure 4 19 Configure Graph Window Since up to six parameters can be dis played at the time the y axis label must be chosen to suit the configured parameters This is achieved by selecting the appropriate radio button adjacent to the parameter list box in the data control bar 4 3 4 3 Control of Data Display The graphical display of parameters has a specified data length default 60 seconds but very often an experiment may last considerably longer MyoView provides a facil ity to scroll back through the display using the slider button I on the control bar Having activated the scroll func
104. www dmt dk sales dmt asia com www dmt usa com support dmt dk or sales dmt dk support dmt asia com support dmt usa com or sales dmt usa com User Manual vers 3 2 Trademarks Safety The Pressure Myograph System has been designed for use only in teaching and research applications It is not intended for clinical or critical life care use and should never be used for these purposes nor for the prevention diagnosis curing treatment or alleviation of disease injury or handicap e Do not open the unit the internal electronics pose a risk of electric shock e Do not use this apparatus near water e To reduce the risk of fire or electric shock do not expose this apparatus to rain or moisture Objects filled with liquids should not be placed on the apparatus e Do not block any ventilation openings Install in accordance with the manufacturer s instructions e Do not install near any heat sources such as radiators heat registers stoves or other apparatus that produce heat e Only use attachments and accessories specified by the manufacturer e Unplug this apparatus during lightning storms or when unused for long periods of time e This apparatus must be grounded e Use a three wire grounding type cord similar to the one supplied with the product e Do not defeat the safety purpose of the polarized or grounding type plug A polarized plug has two flat blades one being wider than the other A grounding type plug has two blades and a
105. y e Press Enter to change between calibration of pressure transducers force transducer or pH in the calibrate line e Use the numeric keys to change the reservoir temperature setting and press Enter to confirm e Enter the specific access code and press Enter to go to the submenu The Setup Menu has the following functions Info F11 F4 Press the Info key and the display shows the F key op tions in the Setup Menu Press Info again to return to the Setup Menu or one of the F keys to proceed with one of these options Press Fi to proceed to the Main Menu Pressure transducer calibration Choose Press in the calibrate line and press F4 to enter the window shown to the right Apply a pressure of 50mm Hg on the Pressure Myo graph using the Big Ben pressure manometer When the relative readings at the bottom line of the window appears stable press F3 to proceed Apply a pressure of 125mm Hg on the Pressure Myo graph using the Big Ben pressure manometer When the relative readings at the bottom line of the window appears stable press F3 to proceed Apply a pressure of 200mm Hg on the Pressure Myo graph using the Big Ben pressure manometer When the relative readings at the bottom line of the window appears stable press F3 to proceed The calibration is now finished The readings in the two bottom lines are the calibration parameters stored in the internal memory
106. y menu item from the pop up menu on the Sequencer Control Bar This action will produce a dialogue box containing the features of the given entry DMT Pressure Myograph System Model 110P amp 111P Sequencer gt wj 0 Initial sal 1 Ince Cur sl e 2 ncre 3 Waitt JE Opera 5 watb E Retur Add entry 1 20 0 30 Remove entry 0 An Clear all entries ED An Load sequence 0 An Save sequence 120 An Start sequencer 0 ap Start at Pause sequence Continuous loop Figure 4 20 Sequencer Control bar Adjust concent Each entry has a number name description annotation and a series of target set tings for the Pressure Myograph System Target values for the pressures and tem peratures can be given a time limit for their achievement duration or the system can be instructed to wait by selecting the appropriate tick boxes until the targets are reached before progressing to the next entry An entry may also be configured to pause the sequence by selecting the Wait for user tick box When the sequence is executed the protocol will pause whenever an entry is reached that has been instructed to Wait for user This action will cause the appearance of a message box containing the text entered in the Prompt area This feature is useful when requir ing a manual intervention during the protocol Instructions for the intervention may be recorded in
107. y the fol lowing characteristics fig 7 3 e The branch points of arteries are V shaped whereas those of veins are more U shaped e The arterial wall contains a thick layer of smooth muscle cells compared to the vein wall which only contains a single or a few layers of smooth muscle cells The histological difference is clearly visible in the stereomicroscope e If you still have difficulty and the vein and artery still contain some blood then try to move the blood forward by very gently squeezing the vessels with a for ceps In the artery the blood will run back quickly whereas in the vein the blood will run back very slowly if it even does so Note it is important that you per form this on vessels other than those you will use as this procedure damages the vessels User Manual vers 3 2 Chapter 7 81 Figure 7 3 Distinguishing between artery and vein 8 Dissect away the vein using scissors to cut the adipose and connective tissue between the artery and vein One method is to cut the vein in one position and af terwards gently to pull the vein away from the artery In this way a fine membrane of connective tissue becomes visible between the adipose tissue and the artery Carefully cut the fine membrane to remove the vein and adipose tissue while avoiding any direct contact between the scissor and artery fig 7 4A B Figure 7 4 Removal of vein 9 Clean the artery by removing any remaining adipose or connective tissu

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