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1. a axararnnnannnnununnnnunnnnnnnnurnnnnnunnnnnnunnnnnnnnnnnrnnnnnnene 40 4 3 Pressure Myograph Maintenance and Cleaning cesses 40 4 3 1 Pressure Myograph Chamber Pipes sssvasauitwendutesumiu beh epe ue renee psu eR cune EE Us 40 4 3 2 Force TFANS UG CF EE Um 40 4 3 3 Pressure Transducer and Glass Cannulas nnnnnrnnnnnnnrnnnnnnnnvnnnnnnrnnnnnnnrennnnnnnnnnnnn 41 5 3 4 Pressure Regulator Cleaning wecencscwssnssdacesesanactbeaseniorsnnedoosadncancacozeaenianedesnaetentosoesaes 41 4 3 5 ETEN NNN 42 Chapter 5 Myograph Accessories and Spare Parts AL 5 1 General Myograph Equipment suernassnnnnnnnnnnnnnnnnnnnnnnnnnunvnnnvnnnnnnnnnnnunnnnsnnnnnnnnnnnenunenene 44 5 2 Pressure Myograph System Accessories assususasunnnnnnnnvnnnnnnnnnunnnnnunnnnnunnnnnunsennnunnnene 44 5 3 Pressure Myograph System Spare Parts rvansunnnnnnnnnnnnunvnnnnnnnnnunnnnnunnnnnnnnnnunnevnnnunenene 45 Chapter 6 Getting Started arranrnannnnnnnnnnannnnnnannnnnnnnnnnnnnnennnnnnennnennennnennennnennnn 46 6 1 Dissection Protocol for Small Mesenteric Arteries rrrrrnrnnnnnrennnnnnrennnnnnvvnnnnnnrennn 46 6 2 Mounting Protocol for Small Arteries ccccccssseeeecesseeeceeaseeeesaseeeeeeaseeeseaeseeseeas 49 Appendix 1 Terms of Warranty anreaanvnannnanenannnnnnnnnennnennnnnnnnnanennnennnennnennnennnnnn 53 Appendix 2 Service Check nnnnnnnnnnnnnnnnnnnnnnnnnnnonnnnnnnnunnnnn
2. Peristaltic pump pH O 14 0 C 50 C 15ul min to 1500ul min 2 5 50 rpm for superfusion of the chamber DMI Confocal Pressure Myograph System Model 120CP Appendix 6 Fuse Changing 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 59 60 DMT Confocal Pressure Myograph System Model 120CP DANISA M10 Texc TL rCOL Eyr DMT A S Skejbyparken 152 DK 8200 Aarhus N Denmark Tel 45 87 41 11 00 Fax 45 87 41 11 O1 www dmt dk Support dmt dk or sales dmt dk DMT Asia Everwin Gardens Rm 502 Block B 521 Wanping Nan Lu Shanghai 200030 China Tel 86 0 21 64869685 Fax 86 0 21 64280591 salesQdmt asia com Support dmt asia com DMT USA Inc 420 Beach Rd 204 Sarasota FL 34242 USA Tel 1 770 612 8014 Fax 1 678 302 7013 www dmt usa com support amp edmt usa com or sales dmt usa com
3. ing the mechanical cleaning with a swab stick DMT Confocal Pressure Myograph System Model 120CP Remove the ethanol solution and wash a few times with double distilled water Incubate the myograph chamber with an 8 acetic acid solution for 10 minutes and continue the mechanical cleaning with a swab stick Wash the myograph chamber several times with double distilled water User Manual vers 3 2 Chapter 4 43 44 Chapter 5 Myograph Accessories and Spare Parts This chapter contains a complete register of equipment needed to set up a basic Confocal 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 need ed for a basic myograph setup at very competitive prices Contact DMT Sales Depart ment for further product information and prices on any of the products listed below 5 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 ltem DS 1000 DMT recommends Geuder G 19745 8 cm straight trabeculum scissors e Dissection and Mounting For
4. 750 CaCl 2H 0 147 02 2 50 9 200 Solution 2 Chemical MW g mol Conc mmol l Conc g l MgSO H 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 3 2 24 0 03 0 250 1 Dissolve the chemicals for solution 1 in about 800m 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 solution 1 25 using double distilled HO 4 Add 1 091 g l Glucose 2 100 g l NaHCO 5 Aerate the solution with carbogen 95760 5 CO for at least 20 minutes If necessary wait further for the pH of the buffer to reach pH 7 4 DMI Confocal Pressure Myograph System Model 120CP Appendix 1 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 Lim
5. Left Glass Cannula Horizontal positioning of the left glass cannula is adjusted by carefully loosening screw D fig 4 1 Vertical positioning of the left glass cannula is adjusted by care fully loosening screw E fig 4 1 Longitudinal positioning of the left glass cannula is adjusted by gently loosening of screw F fig 4 1 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 4 1 Vertical positioning of the right glass cannula is adjusted with the micropositioner C fig 4 1 Longitudinal positioning of the right glass cannula is ad justed by gently loosening of screw G fig 4 1 Note Be careful not to damage the tip of the glass cannulas during the general adjustment 4 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 4 2 and fig 4 3 MT Fr Figure 4 2 Three way valve flow settings at P1 i User Manual vers 3 2 Chapter 4 29 Figure 4 3 Three way valve flow settings
6. 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 valve will make it very difficult to control the pres sure and make the pressure readings unreliable 4 1 5 Replacement of Glass Cannulas The mounted glass cannulas are easily replaced with new ones by carefully loosening the two screws F and G in fig 4 1 p 28 The recommended size for the new glass can nulas is 16mm for the glass cannula connected to P1 right side and 14mm for the glass cannula connected to P2 left side 36 DMI Confocal Pressure Myograph System Model 120CP 4 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 4 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 Confocal Myograph Unit are properly connected thro
7. Rec 2 hi Rec 3 Rec 3 lo Rec 3hi Info Rec 4 gt Temp Change menu Rec 4 lo 3 9 Rec 4 hi 2 BO Cal pressure Info Change menu Change menu Start pres Ctrl Start pres Ctrl Zero force Condensed menu F4 Near HASAN Far 2 7 mN Act 26 5 Set 37 0 Heat OFF pH 7 00 Setup Options Menus Setup Options Menus 7 r No 7 Comm Menu Serial mode RS485 Myograph No 7 pH Menu Offset buffer N A 0 Slope buffer 4100 Temp comp Auto Manual temp 37 C F4 Calibrate Pressure Apply 50 mm Hg Push F3 when stable PI 3614 P2 3533 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 P14956 P2 4933 F3 Calibrate Pressure DONE Push Esc 36148 42527 49565 35334 42223 49332 Figure 3 1 Menus in the 120CP Myo Interface version 3 2 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 ESO F3 Calibrate pH Slope Slope buffer 4 00 Temp auto 22 4 c pH output 177mV F3 Calibrate pH OFFSET Offset 7 2208 Slpoe 26 4 Chapter 3 21 22 3 1 1 Genera
8. a real time flow graph showing flow rate development over time User Manual vers 3 2 Chapter 2 19 20 2 3 2 5 Connecting Oxygen Supply Connect the oxygen supply pipe on the Confocal Pressure Myograph Chamber cover to an adjustable oxygen supply 2 3 2 6 Connecting Suction Device Connect the large pipe on the Confocal Pressure Myograph Chamber cover see fig 2 3 p 18 to a vacuum pump via a suction bottle and a vacuum valve as illustrated in fig 2 1 p 15 The internal diameter of the silicone tube used for this connection must be 2mm A 2mm tube is not included in the myograph system DMI Confocal Pressure Myograph System Model 120CP Chapter 3 The Pressure Myo Interface Chapter 3 is a complete manual for the Confocal 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 Model 120CP Version 3 2 Setup Menu Calibrate Press Resovoir STAGE MainMenu e Inlet i Rec T 64 4 mm Hg P2 Outlet i Rec ilo 55 3 mm Hg Rec 1 hi Status K Flow i 2 2X9 6 mim H sec Rec 2 lo 10 15 mN Ree 2 Option 0
9. at P2 30 4 1 3 How to Read a Millimetre Micrometer Sleeve Scale Timble Scale kd Figure 4 4 Overview of a Micrometer Actual reading 20 00mm Sleeve Scale The micrometer sleeve scale has a total length of 2bmm divided into fifty equal parts Each part of division above the horizontal line represents 1 mm 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 Each divison on the timble scale is 10um If the timble scale is between two lines an approximated figure between O and 10 um must be read Reading Example 1 Enmmmm 1 Note that the thimble has stopped at a point be yond 10 on the sleeve indicating 10000um 2 Note that there is no mark completely visible be tween the 10 mm mark and the thimble Figure 4 5 Example 1 Reading 10380um 3 Read the value on the thimble corresponding to the intersection with the horizontal line on the sleeve A Reading on sleeve 10000um B No additional mark visible 00000um C Thimble reading 380um Total Reading 10380um DMT Confocal Pressure Myograph System Model 120CP Reading Example 2 1 Note that t
10. 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 sealing 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 4 3 3 Pressure Transducer and Glass Cannulas To prevent the pressure transducers and the tubing inside the Confocal Pressure Myograph unit from being blocked by buffer salt deposits after an experiment DMT recommends that the system be flushed using the following procedure 1 Disconnect the Pressure Regulator from the Confocal Pressure Myograph Unit Keep the Confocal Pressure Myograph Unit connected to the Myo Interface to en able 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
11. in the Status line by choosing ON OFF Info Change menu Start pres Ctrl Zero force Condensed menu FA P1 141 Piset 140 P2 129 P2 set 130 EROM T 20 7 T set 37 Pressure Menu P1 Inlet 90 mm Hg P2 Outlet 80 mm Hg Status 2 OFF Flow ON mm Hg sec 10 User Manual vers 3 2 Chapter 3 23 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 automati cally be set equal to the P2 value Also if a P2 value is en tered which is larger than the P1 value then the P2 value will automatically be set equal to the P1 value A setting where P1 P2 activates the No Flow function in the Pressure Regula tor as described in Chapter 4 2 2 p 38 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 Info key and the display shows the F key op tions in the Pressure Menu Info i Change menu Press Info again to return to the Pressure Menu or Start pres Ctrl one of the F keys to proceed with one of these op tions F1 Press F1 to proceed to the Sequencer Menu F2 Press F2 to start the pressure control programme 3
12. 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 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 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 UL81 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 User Man
13. 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 3 4 Pressure Regulator Cleaning To clean the buffer circuit port no P2 and 1 on the Pressure Regulator follow the procedure described in Section 4 1 4 3 34 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 Il 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 4 41 42 4 3 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 L 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 Remove the acetic acid and wash the myograph chamber and glass cannulas several times with double distilled water If any kin
14. the value of the offset buffer first a e buffer solution which is always 7 00 F3 Calibrate pH Slope The second line shows the temperature of the buffer solu Sepe 0 tion The temperature is an important parameter in the ce 00 calibration formula and is obtained automatically by plac ing the myograph temperature probe in the buffer solution The third line shows the output from the pH probe as raw Sor Offset 02208 data from the A D converter Sipe 264 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 relative 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 DMT Confocal Pressure Myograph System Model 120CP 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 Myograph serial identification A B Cor D Myograph A In th
15. which may dam age the mounted vessel when re establishing the flow DMI Confocal Pressure Myograph System Model 120CP Inlet to P1 on Pressure Myograph unit Air In Max 1 Ato Reduction Valve Manometer 0 5 Bar Outlet from P2 on Pressure Myograph Unit Relief Valve 0 8 Bar P3 No Flow Valve vene M Valve 2 Valve 3 Valve 4 JL ie Air Out a Buffer Flow Needle Valve Calibration Valve Figure 5 12 Pressure Regulator Diagram Air Flow User Manual vers 3 2 Chapter 4 39 4 2 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 4 12 p 39 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 4 12 also shows the importance of opening the calibration valve after a pressure calibration If the calibration valve is closed before running an ex periment pressure will build up and damage the mounted vessel because the buffer is unable to return to the left buffer bottle 4 3 Pressure Myograph Maintenance and Cleaning The Con
16. 0 Silicone Tubing at Standard Sizes for the Pressure Myograph System Item ST 1000 User Manual vers 3 2 Chapter 5 45 Chapter 6 Getting Started Chapter 6 contains some practical protocols and procedures describing how to quickly get started using the Confocal Pressure Myograph System The chapter mostly addresses users who are not yet familiar with the pressure myograph technique How ever it may contain some hints that will also be useful for the experienced myograph user 6 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 iseuthanized 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
17. 1 5 Recorder Menu The Pressure Myo Interface has four analogue output ports at the front panel for connection to a data recording system oco NR ec gt ess Each of these output ports can be programmed to represent Re 1lo 0 i i i i Rec 1 hi 22 0 inlet pressure reading outlet pressure reading longitudinal S CN Hr force reading temperature or pH meter The default settings h on are Rec 3 gt Force i Rec 3 lo 200 Rec 3 hi 100 Rec 4 gt Temp P1 Inlet pressure output Rec 4lo O Rec 4 hi 2 50 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 O V lo and 1 V hi DMI Confocal Pressure Myograph System Model 120CP 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 display Press Enter to change between calibration of pressure transducers force transducer or pH in the calibrate line 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 g
18. 2 Pressure Myo Interface e Pressure Myograph Unit connection cable with 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 3 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 4 Riester Big Ben Pressure Manometer 5 Accessories e Allen key e Spare screws e Small screwdriver e Suture nylon wire 3 metres e 1tube of high vacuum grease e 1 tube of grease for linear slides e Silicone tubing internal diameter 1mm 25 metres 6 Hardware Software amp Manuals e User manual Confocal Pressure Myograph System Model 120CP e Cleaning Procedure User Manual vers 3 2 Unpacking the myograph system 11 12 Chapter 1 System Overview 1 1 Confocal Pressure Myograph Unit Horizontal Adjustment Screws r TER T 2 j I Outlet Pressure Perfusion Inlet Pipe Transducer Connection Pipe dE Inlet Valve 25 Pin Serial Port for Commu nication with Myo Interface OOS OT OK Outlet Valve Micropositioner for Vertical Adjustment of Glass Cannulas Force Transducer Pin si Fixat
19. 21 64280591 Fax 1 678 302 7013 sales dmt asia com support dmt asia com www dmt usa com support dmt usa com or sales dmt usa com User Manual vers 3 2 Trademarks Introduction The confocal pressure myograph is a system specifically for use with laser scanning microscopes LSM and other high magnification imaging studies where intracellular processes within the smooth muscle cells or endothelial cells of the intact pressu rized vessel wall are studied Intracellular ion concentration such as calcium and traf ficking of fluorescent probes tags or proteins can be studied in this system in small vessels diameter gt 60um The conical open bath design allows easy access for high magnification high numeri cal aperture lenses used in inverted microscopes It also allows access for direct immersion lenses used in upright confocal microscopy Further the top part of the myograph can be tipped which facilitates maintenance and eases the mounting procedure Using angled custom mounting supports designed specifically to allow very precise vertical positioning an isolated blood vessel can be positioned directly above or on top of the chamber window permitting use of lenses with working distances as low as 10Qum on an inverted LSM The intravascular pressure and the pressure gradient along the vessel can be con trolled precisely and longitudinal force in the segment is measured continuously A heating system with electronic feedback maintai
20. 4 10 2 Connect the Big Ben Pressure Manometer to the 3 way valve at P1 on the Pres sure Myograph Unit as illustrated in fig 4 10 3 On the upper right side of the myograph a small stainless steel pipe is placed It is connected to the right glass cannula via a small silicone tube Carefully discon nect the silicone tube from the stainless steel pipe B 4 Carefully disconnect the silicone tube between the outlet connecting pipe and the outlet pressure transducer connecting pipe C fig 4 11 5 Connect the enclosed resistance tube shunt to the outlet connecting pipe A and the stainless steel pipe B as illustrated in fig 4 11 Figure 4 11 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 P1 now comes from the pressure manometer See fig 4 3C p 29 and fig 4 1 p 28 f 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 Turn on the Pressure Regulator and the Myo Interface Use F1 to go to the Setup Menu in the Myo Interface Bring the calibrate line to the top of the display by usin
21. 6 2 Removal of adipose tissue around the area of interest f 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 by the fol lowing characteristics fig 6 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 f 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 6 47 48 Figure 6 5 A B and C Cutting free the artery to be studied DMT Confocal Pressure Myograph System Model 120CP 6 2 Mounting Protocol for Small Arteries This section shortly describes the basic technique of mounting small vessels in the Pressure Myograph Figure 6 6 Mouting of vessel in the Confocal Pressure Myograph Chamber 1 Fill the Confocal Pressure Myograph Chamber with cold PSS and transfer the isolated ves
22. 9cm in diameter coated with a 5 mm thick layer of Sylgard at the bottom to hold the fixing pins Immedi ately fill the Petri dish with cold PSS well pre bubbled with carbogen see section 6 3 p 86 The dissection 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 6 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 DMI Confocal Pressure Myograph System Model 120CP Second Branch Figure 6 1 Branch of the mesenteric arteries 6 Use high quality forceps and ocular dissection scissors see Chapter 5 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 6 2A B Figure
23. Confocal Pressure Myograph System Model 120CP User Manual Version 3 2 DANISH MYO T HnuNOoL0G DANISA M10 TEaHnNOEolDdDGt DMT Confocal Pressure Myograph System Model 120CP Pressure Myograph Systems Model 120CP 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 Confocal Pressure Myograph Systems Model 120CP Version 3 2 Document Number 120CP 001A No part of this document may be reproduced by any means without the prior written permission of DMT A S Skejbyparken 152 DK 8200 Aarhus N Denmark Tel 45 87 41 11 00 Fax 45 87 41 11 01 www dmt dk supportedmt dk or sales dmt dk DMT Copyright 2006 DMT A S M DANISH MO TECHNOL LC OG DMT Asia Everwin Gardens Rm 502 Block B 521 Wanping Nan Lu Shanghai 200030 China DMT USA Inc 420 Beach Rd 204 Sarasota FL 34242 USA Tel 86 0 21 64869685 Tel 1 770 612 8014 Fax 86 0
24. P Fig 2 1 is a diagram of the complete set up for the Confocal Pressure Myograph Sys tem Model 120CP The diagram includes optional equipment as a peristaltic pump to enable superfusion flow and pH probes for measuring pH in the superfusion buffer Also the diagram include the DMT Flowmeter Model 161FM The Flowmeter enables flow measurements in the range of 15 1500ul min Contact the DMT Sales Department for further product information and prices on any of the optional products shown in fig 2 1 User Manual vers 3 2 Chapter 2 15 16 2 2 Setting up Step by Step Note If you have purchased a computer from DMT in conjunction with your Confocal Pres sure Myograph System the Euresys Picolo frame grabber PCI card has already been installed Follow the procedures in sec 2 2 1 to set up the Confocal Pressure Myo graph System 2 2 1 Setting up the Complete Confocal Pressure Myograph System This section describes how to connect the cables in the Confocal Pressure Myograph system as illustrated in fig 2 1 Note Before proceeding with the connection procedure make sure that the Myo Interface the Pressure Regulator the Microscope and the PC are switched off 1 Confocal Myograph Unit Myo Interface Connection Connect the Confocal 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 Confocal Pressure Myograph Unit 2 Myo Interfac
25. R UK uma dRkre UG iN EnFURihnwskUE Ud IRV ndn Ups rS AD 13 ESPEN recreating EE 14 Chapter 2 Setting Nuvel 15 2 1 Confocal Pressure Myograph System Model 1200P i nnnnvnnnnnrrnnnnvnnnnnrennnnrvnnnnsnenn 15 22 SONNE UD SED OY SEC sassen EEEO EEE 16 2 2 1 Setting up the Complete Confocal Pressure Myograph System 16 2 3 PN 18 2 3 1 The First Weight and Pressure Calibrations wisecccnsaceccasecaicatenencdescsveceassascensccsseacseca 18 2 3 2 External Confocal Pressure Myograph Unit Connections 18 2 3 2 1 Connecting Perfusion Flow asses sesazasruzesugs vEiunaspuutuac pw tuia a ac FU Sav d Reke PEE n RN RDUM E adu ERE 18 2 3 2 2 Extra Perfusion In Outlet nere EE ai vau LENE OH ssp ud e DUM ETE MUE 19 2 3 2 3 Connecting Superrusion Flow VPUONA Le s skcszass essen ra eR RS Dee Fa ennen tapP ETE IqEs rep DREFE 19 2 3 2 4 Connecting Flowmeter Model 161FM Optional 19 2 3 2 5 Connecting Oxygen SUD DIY waadasavmeacccvccanpastcisgancedasautneesxanviceasnasiannguavatctinsttatevsatesens 20 2 3 2 6 Connecting TND Neeme 20 Chapter 3 The Pressure Myo Interface nnrvannnnnnnnnnnnannnnnnnannnnnnnnnennnennunnnnennnnn 21 S VIO IE CF aC IVa UO EE NE aeo tu iesu e 21 odd ee ral KEY FUNCIONS NE ge 22 3 1 2 Turn on Message eek 22 Sula NN 22 S T PEN 23 TR 24 3 LO SCUD MENU en 25 Chapter 4 Pressure Myograph Manual eanrnnnnnnnnnnnnnnnnnnnnnnnnonnnnnnnnnnen
26. T A S Skejbyparken 152 8200 Aarhus N Denmark hereby declares its responsibility that the following product Confocal Pressure Myograph System Model 120 CP 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 Electronic 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 User Manual vers 3 2 Certificate of Conformity Contents FRACS IIA ITI Tm 3 Hg al OG GIO c 4 SUN 5 EET ELLE EEG 6 ABDDPFOVAIIS EE EE NE rer 6 Certificate of Conformity nrnnnnnnnnnnnnnnnnnnnnnnannnnnnnannnnnnnnnnnnnnnnnnnnnnnnnnnnnnennvennnennnene 7 About tnis Manual soncicesnsncccsenanssnsunvscaneeascnedennsvacnssnesdavsnstentandnenssuvacsssecnteevesensneen 10 Unpacking the myograph system nnenavvnnnnnnnnnnnnnnnnnnnnnnunnnnnnnnnnnnnnnennnnnnennnennennr 11 Chapter 1 System Overview rnnnnnnnnnnnnnnnnnnnnnnnannnnnnannnnnnnnennnnnnennennnennnnnnennnnnnen 12 T Confocal Pressure Myograph TIE een 12 1 2 Pressure Myo Interface ausetesms e deiwixkastcac un rua RU vA
27. bration buf fers is measured with a thermometer and entered manually in the manual temperature line Press F1 to return to the Setup Menu User Manual vers 3 2 Chapter 3 27 Chapter 4 Pressure Myograph Manual Chapter 4 contains a complete explanation to all mechanical features of the Confocal Pressure Myograph System This includes how to use adjust and separate the Confo cal Pressure Myograph Unit 4 1 The Pressure Myograph Unit 4 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 Confocal Pressure Myograph Unit The vertical micropositioner fig 4 1 B is used to adjust the vertical position of the right glass cannula and thereby the vertical posi tion of the mounted blood vessel The horizontal micropositioner fig 4 1 C is used to define the length between the two glass cannulas and thereby the horizontal stretch of the mounted blood vessel F G Figure 4 1 Adjustment of glass cannulas DMT Confocal Pressure Myograph System Model 120CP 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
28. ceps ltem 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 LCD Water Bath including Heater DMT recommendes Julabo 5L e Dissection Petri Dish DMT recommends a 9 cm glass Petri dish coated with a 5 mm Sylgaard poly mer layer 5 2 Pressure Myograph System Accessories This section contains a list of special accessories available for the Pressure Myograph System Model 120CP 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 Peristaltic Pump for Superfusion Flow 5 3 Pressure Myograph System Spare Parts This section contains a complete list of standard available spare parts for the Pres DMI Confocal Pressure Myograph System Model 120 CP sure Myograph Model 120CP For parts not listed in this section or for special parts which need to be made customised please contact DMT for further information Glass Cannulas Item GC 1000 Suture Wire Item SW 2000 Calibration Kit Item CK 345110 Chamber Cover Item CC 1110120 Force Transducer Item FT 1110120 Pressure Transducer Item PT 1116115 High Vacuum Grease to Seal Force Transducer Pin hole Item HVG 100
29. control To turn on heating move the Heat line to the top of the display and use Enter to switch between on and off DMI Confocal Pressure Myograph System Model 120CP The Main Menu has the following functions Info Press the Info key and the display shows the F key F1 F2 F3 F4 options 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 Press F1 to proceed to the Pressure Menu Press F2 to start the pressure control programme The settings of the pressure control programme are adjusted in the Pressure Menu Press F3 to zero the longitudinal force reading 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 setting 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 control 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 activated or deactivated
30. d 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 To remove more resistant or toxic 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 HNO solution for about 15 minutes Wash the myograph chamber and glass cannulas several times with double dis tilled water Important Notes Be very careful using step 3 and 4 repeatedly times as strong reagents can cause extreme damage to the myograph unit Be very careful not to damage the glass cannulas during the cleaning procedure 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 E Incubate the myograph chamber for 30 minutes with 20ul of a 2mM T 1210 Tetrakis 2 pyridylmethyl ethylenediamine solution dissolved in double distilled water Use a swab stick to mechanically clean all the affected surfaces during the last 15 minutes of the incubation period Wash the myograph chamber several times with double distilled water Incubate the myograph chamber with 96 ethanol for 10 minutes while continu
31. damage or loss of the myograph system In most cases an additional insurance coverage is needed User Manual vers 3 2 Appendix 3 55 Appendix 4 Calibration of Eyepiece Reticule Principles of Ocular Calibration The purpose of calibrating the eyepiece reticule is to determine a conversion factor allowing the microscope to be used for measuring vessel segment lengths mounted in the wire myograph 60 70 30 90 iu 5 Several types of eyepiece reticules are available for such a purpose The most simple and yet very useful type is a horizontal scale as illustrat ed in Figure A4 1 0 3 Dm m3 40 The basic principle is to use the eyepiece reti cule typically consisting of 50 100 divisions to measure the length of an object in terms of re ticular divisions spanned by the object Having Figure A4 1 Horizontal eyepiece reticule the conversion factor specific for the eyepiece scale reticule and used magnification the length of the object in mm is easily calculated All reticules need to be calibrated in order to determine the specific conversion factor characteristic for the particular eyepiece reticule and the used magnification For such purpose a special devise called a stage micrometer is needed A stage microm eter is simply a microscope glass slide having a scale engraved on the surface A typi cal micrometer scale is 2 00 mm long engraved with divisions of 0 01 mm equalling 10 um per division However micro
32. e 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 cho sen 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 a special code is required Please contact DMT for further information Offset buffer pH value of offset buffer Slope buffer pH value of slope buffer Temp comp AUTO or MANUAL temperature compensation T pH Menu Manual temp Manual temperature setting Offset buffer Slope buffer Temp comp The pH submenu contains the settings of the pH calibration n Sea procedure Offset buffer and slope buffer are the pH values 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 cali
33. e 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 3 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 Confocal 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 4 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 DMI Confocal Pressure Myograph System Model 120CP 5 Turn on the Power Turn the main power to the Myo Interface and the Pressure Regulator on at the power switch and the Microscope Turn on the computer and the Confocal Pres sure Myograph System is now ready to be used Note A detailed guidance in how to connect the Confocal Pressure Myograph System for an experimental se
34. ector REGULATOR RS232 MYOGRAPH e i 3 e Connection Port for con Connection Port for Pres trol of Pressure Regulator sure Myograph Unit RS 232 Port for Serial Connection to PC Connection Port for pH Meter optional USB Dal EE EE e 12V DC Port Figure 1 3 Pressure Myo Interface Back User Manual vers 3 2 Chapter 1 13 14 1 3 Pressure Regulator Pressure and Perfusion Buffer Inlet and Outlet pipes 6 Pressure Transducer Calibration Valve Pressure Valve Activity Indicators Figure 1 4 Pressure Regulator Front Main Pressure Inlet e 115 230 V 50 60 Hz Automatic Voltage Connection Port for the Pressure Myo Interface Selector e e l Interface Pressure inlet ON OFF Switch M Inlet Pressure Manometer Power Connector e Figure 1 5 Pressure Regulator Back DMT Confocal Pressure Myograph System Model 120CP Chapter 2 Setting up 2 1 Confocal Pressure Myograph System Model 120CP pH Meter optional Peristaltic Pump optional ps Myo Interface Rear Panel Connection Cable with temperature probe PC Data Acquisition and Sofware Optional for 120CP FlowMeter 161FM Pressure and i optional Oxygen Supply DMT Microscope Oxygen Superfusion Supply Buffer Vaccum Pump Sucction Waste Bottle Bottle Outlet Inlet Pressure Regulator Rear Panel Figure 2 1 The Complete Confocal Pressure Myograph System Model 120C
35. essure Myograph Unit use the silicone tube internal diameter 1mm delivered with the Confocal Pressure Myograph System Connect the perfusion inlet at P1 on the Confocal Pressure Myograph Unit to the P1 outlet on the cap of the buffer bottle Connect the perfusion outlet at P2 on the Confocal Pressure Myograph Unit to the P2 inlet in the Pressure Regulator Unit see fig 2 1 p 15 DMI Confocal Pressure Myograph System Model 120 CP Note I To fill the tubing and avoid air bubbles mount the Resistance tube shunt as de scribed in Chapter 4 1 4 3 p 34 Fill the buffer bottle with 250ml buffer and run the following flush programme P1 150 mm Hg P2 70 mm 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 250ml 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 Confocal Pressure Myograph Chamber as this may cause serious damage to the vessel 2 3 2 2 Extra Perfusion In Outlet The extra perfusion in and outlet on the Confocal Pressure Myograph Unit makes it possible to add special agents or reagents to the perfusion circuit during an experi ment The small handle on top of the valves controls the flow direct
36. focal Pressure Myograph System Model 120CP 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 read carefully and that the instructions are followed at all times 4 3 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 4 3 2 Force Transducer The force transducer is one of the most delicate and fragile components of the Confo cal Pressure 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 4 13 To prevent the buffer from running into the transducer house the hole is filled with high vacuum grease Figure 4 13 Transducer pin hole sealed up with high vacuum grease DMT Confocal Pressure Myograph System Model 120CP As a part of daily maintenance it is very important to inspect the
37. g 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 User Manual vers 3 2 Chapter 4 35 10 11 12 13 Apply a pressure of 50mm 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 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 F4 Calibrate Pressure Apply 50 mm Hg Push F3 when stable Pi 36141P2 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 P14956 P2 4933 F3 Calibrate Pressure DONE Push Esc 36148 42527 49565 35334 42223 49332 Carefully remove the resistance tube shunt and reconnect the tubing to the glass cannulas Open the calibration valve on the
38. he thimble has stopped at point beyond 16 on the sleeve indicating 16000um 2 Note that this time a mark is visible between the 16 mm mark and the thimble indication 500um Figure 4 6 Example 2 3 Read the value on the thimble corresponding to the Reading 16780um intersection with the horizontal line on the sleeve A Reading on sleeve 16000um B One additional mark visible 500um C Thimble reading 280um Total Reading 16780um 4 1 4 Force and Pressure Calibration Section 4 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 4 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 Per t r F sing 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 4 7 User Manual vers 3 2 Chapter 4 31 32 Tra
39. ion Plate for left Glass Cannula e Outlet Connecting Pipe boo ge eFixation Plate for Right Glass Cannula Am PE Sn eae ae MULA 1 00 MARET Silicone Tube connecting Perfusion Silicone Tube connecting Outlet Inlet Pipe and Right Glass Cannula Connecting Pipe with Left Glass Cannula i ERE E e Right Glass Cannula Left Glass Cannula ee e Myo Chamber Window Figure 1 1 The Confocal Pressure Myograph Chamber DMT Confocal Pressure Myograph System Model 120 CP 1 2 Pressure Myo Interface Up and Down Arrow Keys Scroll up and down in the menus If you are at the top of a e i menu use f to change to the previous main menu Analogue Recorder Output Connection to external data acquisition system optional Myo Interface Display e i e F Keys Have different but specific func Numeric Keys Enter data or setting values tions dependent of the active main menu bure Enter Key Confirm entered numeric val INFO Key Push this key in any menu ues or change between available choices to display all specific F key functions e ESC Key Delete incorrectly entered numeric values z In some submenus used to return to main menu Figure 1 2 Pressure Myo Interface Front 115 230 V 50 60 Hz Automatic Voltage Selector e pueeeeseneseeeeneeeniee ense tne ensee ense ttov ettet enormen e ON OFF Switch puente nennen nennen nnne nennen nennen e Power Conn
40. ions through the 3 way valves at P1 and P2 as described in Section 4 1 2 p 29 2 3 2 3 Connecting Superfusion Flow Optional Creating a superfusion flow in the Confocal Pressure Myograph Chamber requires an external peristaltic pump which is not part of the standard Confocal Pressure Myo graph System Contact DMT for further information about recommended perfusion pump models specifications and prices The superfusion circuit is connected to the Confocal Pressure Myograph Unit as illus trated in fig 2 1 p 15 2 3 2 4 Connecting Flowmeter Model 161FM Optional The DMT Flowmeter Model 161FM is optional for flow measurements in the range of 15 1500ul min The Flowmeter is easily combined with any DMT Pressure Myo graph System as illustrated in fig 2 1 p 15 Use of the FlowMeter 161FM makes it possible to control intravascular pressure pressure gradient and flow conditions with high accuracy which enables the per formance of in vitro studies on vessel segments under conditions very close to those found in vivo Application of the FlowMeter 161FM also enables measurement of flow rate as a parameter due to changes in vessel structure and function in various physi ological and pharmacological studies The FlowMeter 161FM 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
41. is sta PE ble Press F3 to proceed with calibrating Force 1938 F3 f Carefully place the 2 g weight on the pan as illustrated in st Force fig 4 7 p 34 The force applied on the force transducer pla Fa Men sable Force 2245 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 Calibrate Force DONE Push Esc Force Zero 20315 Force gain 668 3 8 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 MN 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 Confocal Pres sure Myograph System is now ready for use longitudinal force measurements 4 1 4 3 Pressure Calibration Procedure The setup to perform the pressure transducer calibration procedure is illustrated in fig 4 10 Carefully follow the procedure described below to pressure calibrate the Pressure Myograph System Figure 4 10 Illustration of set up for pressure transducer calibration DMT Confocal Pressure Myograph System Model 120CP 1 Connect the Confocal Pressure Myograph Unit to the Pressure Regulator as to perform a standard pressure myograph experiment as illustrated in fig
42. itations 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 for 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 1 53 54 Appendix 2 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 contac
43. l Key Functions P Scroll up and down in the menus Being in the top line of a menu use 7 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 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 120CP contrast setting Version 3 2 Press F1 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 MainMenu Force Longitudinal force reading in mN ae a Temp X Actual temperature probe reading in C 10 15 mN Set temp Heating temperature setting in C T FR Heat Actual heating status ON OFF qun pH Actual pH reading Optional 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
44. le P14252 P2 4222 F3 Calibrate Pressure Apply 200 mm Hg Push F3 when stable P14956 P2 4933 F3 Calibrate Pressure DONE Push Esc 36148 42527 49565 35334 42223 49332 User Manual vers 3 2 Chapter 3 25 For a more detailed instruction to the calibration procedure see Chapter 4 1 4 p 31 Force transducer calibration Chose Force in the calibrate line and press F4 to enter the window shown to the right Calibrate Force Apply no Force Push F3 when stable Force 1938 Make sure that no force is applied on the force transducer and when the relative reading at the bottom line of the window appears stable press F3 to proceed dd re Place 2 g on the pan Push F3 when stable Carefully place the 2 gram weight on the load pan and Force 2245 when the relative reading at the bottom line of the window appears stable press F3 to proceed F3 Calibrate Force m i DONE Push Esc The calibration is now finished The readings in the two Force Zero 20315 i F in 668 3 bottom lines are the calibration parameters stored in the SEE ag 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 4 1 4 p 31 pH calibration Optional Choose pH in the calibrate line and press F4 to enter the z window shown to the right Calibrate pH OFFSET Offset buffer 7 00 Temp auto 22 4 c The first line shows
45. meter glass slides with less fine divisions are also useful for calibrating a stereomicroscope to be used with the wire myograph Ocular Calibration Procedure a Decide which microscope magnification is to be used for the segment length measurements Use the largest possible fixed magnification where the eyepiece reticule scale still covers the whole gap of the myograph jaws b Place the stage micrometer on the microscope stage and focus on it Fit one of the division lines on the stage micrometer to one of the division lines of the reticule scale very precisely While keeping the stage micrometer absolutely fixed on the microscope stage find another position on both scales where the division lines also fit precisely Read the position of the two fit points on both scales and fill in the values in the Ocular Calibration Sheet Repeat the procedure twice DMI Confocal Pressure Myograph System Model 120CP Ocular Calibration Sheet Date jOperator Magnification Frontlens Stage micrometer Reticutar scale 2 fon a mm fam fam av 3 few a mm fam fam av wen mm fa Calculations a Calculate the length between the two positions on the stage micrometer by sub tracting the value of position 1 from the value of position 2 Multiply the length in divisions with the length of each division to get the length in mm b Calculate the length betwee
46. n the two positions on the reticule scale by subtract ing the value of position 1 from the value of position 2 c Calculate the mean length value of both the stage micrometer and the reticular scale d Calculate the conversion factor Mean Stage Micrometer Length mm Mean Reticule scale div User Manual vers 3 2 Appendix 4 58 Appendix 5 System Specifications Specifications Vessel size Vessel alignment Mounting supports Chamber Chamber volumes Chamber material Chamber cover Force range Force resolution Pressure range Pressure resolution Heating Temp range Temp resolution Temp probe Weight calibration Analogue output Digital output Voltage Pressure regulator Pressure range Pressure calibration Fluid capacity Gas superfusion ports Heating Valves Voltage Optional accessories gt 60um Manually X Y amp Z Glass pipettes Single Max 10ml 3ml typical Acid resistant stainless steel With pipes for suction gassing superfusion 5BOMN O 01mN 0 250mmHg 1 0mmHg Built in Ambient temp 50 C 017C Included Semi automatic 1 0V F S Serial interface RS232 RS485 100 to 240 VAC auto 50 60 HZ O 250mmHg requires external pressure source Manual 250ml Built in Built in Safety and reduction 100 to 240 VAC auto 50 60 HZ Enable pH meter on the interface range temp correction FlowMeter range
47. nennnennnn 28 4 1 The Pressure Myograph UNnit nnnnannnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnne 28 4 1 1 Adjustment of Glass Cannulas sisicdeseccccesaincuastnacctucenasensennteabsrancdoieanedaraamedaadecneencenss 28 Cele SVEV Valve GS EE EEE 29 4 1 3 How to Read a Millimetre Micrometer rrnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnennnnsnennn 30 4 1 4 Force and Pressure Calibration xrrvasunnnasunnnnnnunnnnnnnnnnnnnnnnnnnnnnnnunnnnnnnnnnunnnnnnnunvnene 32 4 1 4 1 Principles of Weight Calibration sauene tekst tr LEE ERE ERE E REELLE REE Suas 31 FE Weight Calibration Procedure eee en 33 4 1 4 3 Pressure Calibration Procedure rnannnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnr 34 4 1 5 Replacement of Glass Cannulas a annanunnannnnnnunnanunnnnunnnnunnnnnnnnnunnnnunnnnnnennunennnnennne 36 4 1 6 Checking the Force Transducer scicsnscesccsrisocsaninmechasscoteasedetseinreccdsrsanccanssatdansenivoasones 37 4 1 6 1 Simple Force Transducer UNE ua 37 4 1 7 Checking the Pressure Transducers rrnnnnnnnnnnnnnnrnnnnnnnnnnnnnnnnvnnnnnnrennnnnnrennnnensnnnnn ST 4 1 7 1 Simple Pressure Transducer Check annanannnnanannannnnnnunnnnannnnnnnnnnnnnnunnnnnnnnnnnnnnnunnnne 38 42 MENE TER Ne 38 4 2 1 Control OF Flow and Pressure SCUINGS arvemasse eee KR u aseo E aas 38 4 2 2 No Flow Setting EET EE NE 38 DMI Confocal Pressure Myograph System Model 120CP 4 2 3 Pressure Transducer CalibratiOn
48. nnnennnnnnennnnnnennnnnnennnnnn 54 Appendix 3 Shipping Instructions eere ener enn nnn 55 Appendix 4 Calibration of Eyepiece Reticule 56 Appendix 5 System Specifications cce nee rene 58 Appendix 6 Fuse Changing rnanvannnnnnnnnnnnnnnnnnnennnnnnennnnnnnnnnennnnnnennennnennnnnnennnnnn 59 User 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 Confocal Pressure Myograph System Model 120CP Version 3 2 Chapter 1 provides an overview of the construction and basic features of the com plete Confocal Pressure Myograph System Chapter 2 describes step by step how to set up a complete Confocal Pressure Myo graph System 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 Confocal Pressure Myograph System Chapter 4 is a complete manual to the Confocal Pressure Myograph System The chapter describes in detail how to use and adjust the Confocal Pressure Myograph chamber and finally instructions to daily maintenance of the Confocal Pressure Myo graph System Chapter 5 is a list of equipment needed to set up a complete Confocal Pressure Myo graph System The chapter also contains a list of acce
49. ns bath temperature above ambient temperature The customized chamber cover includes ports for superfusion for rapid draining and filling and for gas supply To facilitate cleaning the chamber is made in stainless steel and is acid resistant When using the myograph on a confocal microscope data acquisition is possible of in and outlet pressure force and temperature Using the analogue outputs or the se rial interface does it either DMT Confocal Pressure Myograph System Model 120 CP Safety The Confocal 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
50. nsducer Arm TA L Figure 4 7 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 FT acting on the force transducer The following two equations describe the forces working in the weight calibration system Gamia Fa IA ub m i4 7 8 SING 2 TL FSM where r is the length of the pan arm F is the force acting on the pan arm when applying the weight F is equal acceleration of gravity times the mass of the weight 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 ER m is 8 SING FLUE 8 sind r SING As the length of the pan arm is 2cm the length of the transducer arm is 4cm the weight is 2g both angles are 900 and the acceleration of gravity is 9 81ms the force acting on the force transducer is F 2cm 2g 9 81ms sin90 4cm sin9O F 9 81 gram m s As 1N is equal 1kg m s F is equal to F 9 81mN DMT Confocal Pressure Myograph System Model 120CP 4 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 wi
51. o to the submenu The Setup Menu has the following functions Info 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 F1 Press F1 to proceed to the Main Menu F4 Pressure transducer calibration Choose Press in the calibrate line and press F4 to enter the window shown to the right Apply a pressure of 50 mm 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 125 mm 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 200 mm 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 of the Myo Interface Press ESC to return to the Setup Menu SetupMenu Calibrate Press Resovoir E 7C Option 2 0 Info Change menu Cal pressure F4 Calibrate Pressure Apply 50 mm Hg Push F3 when stable pi 36120 P2 3533 F3 Calibrate Pressure Apply 125 mm Hg Push F3 when stab
52. ounted vessel DMT Confocal Pressure Myograph System Model 120CP 6 3 Buffer Recipes 6 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 1 17 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 dissolve the EDTA 2 Solution 1 is added to a graduated bottle and the bottle is filled with double distilled HO 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 O 576 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 6 4 User Manual vers 3 2 Chapter 6 51 52 25x Concentrated PSS 1000ml Solution 1 Chemical MW g mol Conc mmol l Conc g l NaCl 58 44 118 99 173 850 KCI 14 56 4 69 8
53. sel segment to the Confocal 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 Confocal Pressure Myograph Chamber in the Main Menu on the Myo Interface The PSS is in the myograph chamber is then gradu ally warmed to 37 C over a period of 15 minutes 6 Place the chamber cover over the Confocal Pressure Myograph Chamber and open for the oxygen supply both to the pressure 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 User Manual vers 3 2 Chapter 6 49 Figure 6 7 MyoView picture of a m
54. ssories and spare parts avail able to the Confocal Pressure Myograph System Model 120CP Chapter 6 contain procedures describing how to get started using the Confocal Pres sure Myograph System This includes a complete dissection and mounting procedure Appendices contain additional information about myograph service shipping instruc tions system specifications serial protocol and fuse changing DMI Confocal Pressure Myograph System Model 120CP Unpacking the myograph system Please take a few minutes to carefully inspect your new Confocal 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 Confocal Pressure Myograph System In case of transport and the original packing material is unavailable please contact DMT Sales Department for advice and packing instructions After unpacking your new Confocal Pressure Myograph System please use the follow ing list to check that the system is complete 1 Confocal 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
55. t DMT for information about prices DMI Confocal Pressure Myograph System Model 120CP Appendix 3 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 preferable 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
56. th 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 myograph unit as illustrated in fig 4 8 Figure 4 8 Weight Calibration Set up 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 4 9 Carefully move the 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 d A Figure 4 9 Illustration of how to fit the balance just behind the glass cannula fixation plate User Manual vers 3 2 Chapter 4 33 34 5 Goto the Setup Menu on the Myo Interface and choose Force to weight calibrate the force transducer Press F4 to start calibration Setup Menu Calibrate Force Resovoir ESTE Option 2 6 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 SE check that the relative force reading in the display
57. the pressure applied on the pressure transducers which prevent them from being dam aged The applied flushing pressure should not exceed 150mm Hg This chapter describes how to check the pressure transducers for any kind of dam age User Manual vers 3 2 Chapter 4 31 38 4 1 7 1 Simple Pressure Transducer Check 1 Ifthe pressure reading on the Myo Interface appears unstable then first check that the Myo Interface and the Confocal Pressure Myograph Unit are properly con nected through the 25 pin grey cable 2 If the pressure reading still appears unstable then perform a new pressure cali bration of the pressure transducers as described in Section 4 1 4 3 p 34 During the pressure calibration monitor the three relative pressure reading values in the calibration menu on the Myo Interface e f the value is 0 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 3001 6250 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 4 2 The Pressure Regulator The Confocal Pressure Regulator controls the flow and pressure applied to the vessel mo
58. tup is described in detail in chapter 2 3 p 20 User Manual vers 3 2 Chapter 2 17 18 2 3 Experiment Setup 2 3 1 The First Weight and Pressure Calibrations Prior to the shipment of the Confocal Pressure Myograph System it has gone through two days of continuous testing including final weight and pressure calibrations How ever in order to ensure that the myograph is working at highest performance DMT recommends that new weight and pressure calibrations are performed before start ing to use the Confocal Pressure Myograph System The weight and pressure calibration procedures are described in detail in Chapter 4 1 4 p 31 2 3 2 External Confocal 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 for connection Superfusion outlet Perfusion inlet e b to vacuum P e d 3 p i i a Extra perfusion Superfusion Pipe for connedction Access hole for Extra perfusion outlet inlet to oxygen supply reagents buffer inlet temperature or pH probe Figure 2 3 External Pressure Myograph connection 2 3 2 1 Connecting Perfusion Flow The perfusion flow is controlled by the Pressure Regulator via the Myo Interface How to control the perfusion flow is described in detail in chapter 3 1 4 p 23 To connect the Pressure Regulator to the Confocal Pr
59. ual vers 3 2 Safety 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 generates 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 DMI Confocal Pressure Myograph System Model 120CP Certificate of Conformity DM
60. ugh 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 4 1 4 2 p 32 During the weight calibration monitor the relative force reading values in the calibra tion menu on the Myo Interface e If the value is O or above 6500 then the force transducer is broken and needs to be changed e fthe reading is between 1 499 or 3001 6250 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 4 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 flushing of the pressure transducers the tubing inside the pressure myograph unit and the glass cannulas is performed with the Myo Interface connected and turned on when using a syringe This enables an exact control of
61. unted in the pressure myograph chamber via settings in the Myo Interface and the MyoView software This Chapter is intended as an overview of the Pressure Regula tor to understand how it works to control flow and pressure settings 4 2 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 4 12 p 39 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 buffer more than 250ml as higher volumes makes precise control of the pressure more difficult 4 2 2 No Flow Setting Activating the No Flow setting in the Myo Interface closes the No Flow Valve in the diagram shown in fig 4 14 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 4 12 Activating P3 serves to prevent a huge pressure to build up in the left bottle with buffer

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