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1. CHAPTER 3 23 CHAPTER 4 CONFOCAL PRESSURE MYOGRAPH SYSTEM 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 Confocal 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 see figure 4 1 B below is used to adjust the vertical position of the right glass cannula and thereby the verti cal position of the mounted blood vessel The horizontal micropositioner See figure 4 1 C below is used to define the length between the two glass cannulas and thereby the horizontal stretch of the mounted blood vessel A B e o eececccc c06060606000000000 0 060 t 9999999999 0 0 0 0 O0 O O 6 9 G Figure 4 1 Adjustment of glass cannulas 24 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL 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 o
2. CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP 2 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP TRADEMARKS PowerLab and LabChart are registered trademarks of ADInstruments Pty Ltd The names of specific recording units such as PowerLab 4 25 are trademarks of ADInstruments Pty Ltd Pentium is a registered trademark of the Intel Corporation Windows Windows 95 Windows 98 Windows ME Windows NT Windows 2000 Windows XP and Vista 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 System Model 120CP Document Number 120CP 002A All rights reserved No part of this manual may be reproduced or transmitted in any form or by any means without the written permission of Danish Myo Technology A S Every attempt is made to ensure accurate information misprints construction and specification changes can occur Danish Myo Technology A S reserves the right to alter change content as required and without any notice Copyright Danish Myo Technology A S DANISH MYO TECHNOL OGS TRADE
3. Chapter 3 The Confocal Pressure Myo Interface ccssssscccsensseccensseecensseeneensseenennsseceennsseeeonsseeneonssseneensseenenssseceennsssesennases 18 SUM Te EIS RES IS emi el S NT RIOT NE n 18 Chapter 4 Confocal Pressure Myograph system manual 1 eec cierre eee esee enean enun nana aane ananas oaa a aan saa u 24 Ad Ihe Pressure Myogcraph UME asesir m 24 ATC eS oe MeO OAL Ol a A E 30 AS Pressure myosraphmalntenance and 32 Appendix 1 Fuse a 34 Appendix 2 System Specifications civisssicccssscccisnssnsnescsssncsusnessepentdreucerancausnndixsessdsnsseuundsssnensunesnicanssseosbanvessunassnuendsnseaniverssusnseaveceues 35 4 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL INTRODUCTION The Confocal Pressure Myograph is a system specifically for use with laser scanning microscopes LSM and other high magnifica tion imaging studies where intracellular processes within the smooth muscle cells or endothelial cells of the intact pressurized vessel wall are studied Intracellular ion concentration such as calcium and trafficking of fluorescent probes tags or proteins can be studied in this system in small vessels diameter gt GOum The conical open bath design allows easy
4. CHAPTER 3 21 Force transducer calibration F4 Calibrate Force Chose Force in the calibrate line and press F4 to enter the window shown to the right 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 F3 Carefully place the 2 gram weight on the load pan and when the relative reading at the cere aetna bottom line of the window appears stable press F3 to proceed Push F3 when stable Force 2245 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 F3 Calibrate Force DONE Push Esc For a more detailed instruction to the calibration procedure see Chapter 4 1 4 Pressure Force rero eee tS Calibration Procedure on page 27 Force gain 668 3 pH calibration Optional Choose pH in the calibrate line and press F4 to enter the window shown to the right F4 The first line shows the value of the offset buffer first buffer solution which is always Calibrate pH OFFSET 700 Offset buffer 7 00 z Temp auto 22 4 c pH output 2 The second line shows the temperature of the buffer solution The temperature is an important parameter in the calibration formula and is obtained automatically by placing the myograph tempe
5. 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 Myograph System Appendices contain additional information about fuse replacement and system specifications ABOUT THIS MANUAL 9 UNPACKING MYOGRAPH SYSTEM Please take a few minutes to carefully inspect your new Confocal Pressure Myograph System for damage which may have oc curred 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 pack ing instructions After unpacking your new Confocal Pressure Myograph System please use the following list to check that the system is complete 1 Confocal Pressure Myograph Unit Force calibration kit Including bridge balance and 2 gram weight Resistance tube shunt Cali
6. 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 Myograph using the Big Ben pressure manometer When the relative readings at the bottom line of the win dow 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 win dow 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 win dow appears stable press F3 to proceed The calibration is now finished The readings in the two bottom lines are the cal ibration 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 4 1 4 Pressure Calibra tion Procedure on page 27 Setup Menu Calibrate Press Resovoir 2 OG Option 2 0 Info Change menu Cal pressure F4 Calibrate Pressure Apply 50 mm Hg Push F3 when stable Pi 3614 P2 3593 F3 Calibrate Pressure Apply 125 mm Hg Push F3 when stable Z5 2288 2 872 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
7. access for high magnification high numerical aperture lenses used in inverted micro scopes It also allows access for direct immersion lenses used in upright confocal microscopy Further the top part of the myo graph 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 100um on an inverted LSM INTRODUCTION 5 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 Do not open the unit the internal electronics pose a risk of electric shock Do not use this apparatus near water 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 Do not block any ventilation openings Install in accordance with the manufacturer s instructions Do not install near any heat sources such as radiators heat registers stoves or other apparatus that produce heat Only use attachments and accessories specified by the man
8. follow the proce dure described below to pressure calibrate the Pressure Myograph System Figure 4 6 Illustration of set up for pressure transducer calibration 1 Connectthe Confocal Pressure Myograph unit to the Pressure regulator as to perform a standard pressure myograph experi ment as illustrated in fig 4 6 above 2 Connect the Big Ben Pressure manometer to the 3 way valve at P1 on the Pressure Myograph unit as illustrated in fig 4 6 above 3 Onthe 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 disconnect the silicone tube from the stainless steel pipe see figure 4 7 B on next page 4 Carefully disconnect the silicone tube between the outlet connecting pipe and the outlet pressure transducer connecting pipe see figure 4 7 C on next page 5 Connect the enclosed resistance tube shunt to the outlet connecting pipe A and the stainless steel pipe B as illustrated in figure 4 7 on next page CHAPTER4 27 Figure 4 7 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 Figure 4 3 Three way valve flow settings at P2 on page 25 and Figure 4 1 Adjustment of glass cannulas on page 24 f Close the calibration valve on the front of the Pressure r
9. 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 CHAPTER4 29 4 2 The Pressure regulator The Confocal Pressure Regulator controls the flow and pressure applied to the vessel mounted in the pressure myograph cham ber via settings in the Myo Interface and the DMT VAS 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 figure 4 8 on next page 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 250 ml AS HIGHER VOLUMES MAKES PRECISE CONTROL OF THE PRES SURE 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 figure 4 8 on next page 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 My
10. the Pressure regulator by turning it counter clockwise IMPORTANT MAKE SURE THAT THE CALIBRATION VALVE IS COMPLETELY OPENED AFTER THE PRESSURE CALIBRATION A HALF CLOSED CALIBRATION VALVE WILL MAKE IT VERY DIFFICULT TO CONTROL THE PRESSURE AND MAKE THE PRESSURE READINGS UNRELIABLE 28 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL F4 Calibrate Pressure Apply 50 mm Hg Push F3 when stable P13614 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 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 Figure 4 1 Adjustment of glass cannulas on page 24 The recommended size for the new glass cannulas is 16mm for the glass cannula con nected to P1 right side and 14 mm for the glass cannula connected to P2 left side 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 th
11. 2003 EMC EMI 7 CERTIFICATE OF CONFORMITY DMT A S Skejbyparken 152 8200 Aarhus N Denmark hereby declares its responsibility that the following product Confocal Pressure Myograph System Model 120 CP is covered by this certificate and marked with CE label conforms with the following standards EN 61010 1 2001 Safety requirements for electrical equipment for measurement control EN61010 1 Corr 1 2003 and laboratory use EN 61010 1 Corr 1 2003 Part 1 General requirements EN 61010 2 101 2003 Safety requirements for electrical equipment for measurement control and laboratory use Part 2 101 Particular requirements for in vitro diagnostic IVD medical equipment Electrical equipment for measurement control and laboratory use EMC EN 61326 2 6 2005 requirements Part 2 6 Particular requirements In vitro diagnostic IVD medical equipment With reference to regulations in the following directives 2006 95 EC SY 550 EEC 8 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL ABOUT THIS MANUAL This manual contains a complete list of procedures describing how to install maintain and get started using the Confocal Pres sure Myograph System Model 120CP Chapter 1 provides an overview of the construction and basic features of the complete Confocal Pressure Myograph System Chapter 2 describes step by step how to set up a complete Confocal Pressure Myograph System Chapter 3 is a complete manual to the Myo
12. 5 1 ec Rec 1 hi 200 Rec 4 P1 Inlet pressure output a 7 m 2 ec 5 Rec 2 P2 Outlet pressure output Rec 2hi 200 Rec 3 Longitudinal force output ae QUEE eo 0 Rec 4 Temperature output Rec 3hi 100 Rec 4 gt Temp 4 P Rec 4 lo 6 The full scale output on the recorder is one volt In the Recorder Menu it is possible to Rec 4hi 50 change the value that corresponds to O V lo and 1 V hi 20 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL 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 e 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 Enter the specific access code and press Enter to go to the submenu The Setup Menu has the following functions Info F1 F4 Press the Info key and the display shows the F key options 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 F1 to proceed to the
13. EDURES IN THE USER MANUAL DMT VAS MANUAL CORE amp ANALYSIS FUNCTIONS TO INSTALL DMT VAS ON YOUR OWN COMPUTER 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 figure 2 1 on previ ous page NOTE BEFORE PROCEEDING WITH THE CONNECTION PROCEDURE MAKE SURE THAT THE MYO INTERFACE THE PRESSURE REG ULATOR 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 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 Interface 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
14. I I DMT Microscope Not included Superfusion Buffer I I I I I I I I I I I I I I I I I I I I Oxygen Supply Pressure and Oxygen Supply Vaccum Pump optional Sucction Bottle Outlet Superfusion buffer Inlet Superfusion buffer Inflow Perfusion buffer Outflow Perfusion buffer Pressure Regulator Rear Panel Figure 2 1 Example of the Complete Confocal Pressure Myograph System Model 120CP Fig 2 1 is a diagram of the complete set up for the Confocal Pressure Myograph System model 120CP The diagram includes op tional 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 161FM The Flowmeter enables flow measurements in the range of 15 1500 ul min Contact the DMT Sales Department for further product information and prices on any of the optional products shown in figure 2 1 above 14 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL 2 2 Setting up step by step NOTE IF YOU HAVE PURCHASED A COMPUTER FROM DMT ALONG WITH YOUR CONFOCAL PRESSURE MYOGRAPH SYSTEM THEN THE DMT VESSEL ACQUISITION SUITE DMT VAS HAS ALREADY BEEN INSTALLED ALONG WITH DRIVERS FOR THE DIGITAL CCD USB CAMERA FOLLOW THE PROCEDURES IN SECTION 2 2 1 BELOW TO SET UP THE PRESSURE MYOGRAPH SYSTEM IF YOU HAVE NOT PURCHASED A COMPUTER FROM DMT FOLLOW THE PROC
15. MARKS 3 CONTENTS Uh CU TN AS ceed cae he ec see nw ne oa ec ee ene see le he es sen 3 DUNE M 5 M 6 EMC 2 7 7 P 8 c ntc 9 Unpacking the myograph system seni oco xvi oER YR UB KEEN 10 Chapter 1 System overview cesi eux S RE EX EDU E E uU BREK VEM E E E EARN EN ERN NEM NNNN FUE KM KEE nnmnnn nnmnnn CR EM RE REN NECEM NENNEN E 11 1 4 Confocal Pressure Myograph piden XR 11 1 2 Confocal Pressure Myodnterfate piesa ita UH A RED a TM dh ota atv DOR Oba NIGRAE a SDN heme Ida NNNM ON DOM NNUS E ON MODI chub 12 L3 PIESSUTE OBI duce cite ccs hc sc atta cal coUS ERN NEN UE 19 Chapter 14 2 1 Confocal Pressure Myograph System Model 1200P os viciisiisas ha raw o pra ai RAE E Iw ce 14 Pare CNS UD SLC SLE NEP 15 Fi EE E T o A LEN 15
16. bration tube Two glass cannulas e Chamber cover Pressure Myo Interface Pressure Myograph Unit connection cable with a temperature probe 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 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 Two 500 ml Schott Duran Bottles incl special designed screw caps Riester Big Ben pressure manometer Accessories e Allen key e Spare screws e Small screwdriver e 3m nylon thread e tube of high vacuum grease e tube of grease for linear slides Silicone tubing internal diameter 1 mm 25 m Hardware software amp manuals e 1 CD with user manual Confocal Pressure Myograph System model 120CP e Cleaning procedure 10 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL CHAPTER 1 SYSTEM OVERVIEW 1 1 Confocal Pressure Myograph Unit Horizontal adjustment screws peser nnne nnne nnns i Outlet pressure transducer connection pipe e 25 pin serial port for commu nication with Myo Interface e 9 9 Outlet valve 5 Force transducer pin mOem x Figure 1 1 The ConfocakPressure Myograph chamber Perfusion
17. ctivated in the Status line by choosing ON OFF IMPORTANT PLEASE NOTE THAT THE P1 INLET PRESSURE CANNOT BE SMALLER THAN THE P2 OUTLET PRESSURE IF A P1 VALUE SMALLER THAN THE P2 VALUE IS ENTERED THE P1 VALUE WILL AUTOMATICALLY BE SET TO A VALUE EQUAL P2 FURTHERMORE IF A P2 VALUE LARGER THAN THE P1 VALUE IS ENTERED THEN THE P2 VALUE WILL AUTO MATICALLY BE SET TO A VALUE EQUAL P1 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 gradi ent The Pressure Menu has the following functions Info Press the Info key and the display shows the F key options in the Pressure Menu Info Press Info again to return to the Pressure Menu or one of the F keys to pro i CUI DIS Start pres Ctrl ceed with one of these options F1 Press F1 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 con nection to a data recording system Each of these output ports can be programmed to represent inlet pressure reading outlet pressure reading longitudinal force reading recorder Men temperature or pH meter The default settings are i A
18. e 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 Myo graph Unit are properly connected through the 25 pin grey cable 2 f the force reading still appears unstable then perform a new weight calibration of the fore transducer as described in 4 1 3 Weight Calibration Procedure on page 26 During the weight calibration monitor the relative force reading values in the calibration 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 If the 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 trans ducers are placed inside the pressure myograph unit to protect them from mechanical damage The pressure transducers are very vu
19. egulator by turning it clockwise IMPORTANT IF THE CALIBRATION VALVE IS NOT COMPLETELY CLOSED IT WILL RESULT IN AN INACCURATE PRESSURE CALIBRATION AND UNRELIABLE PRESSURE READINGS AN SETTINGS DURING AND 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 cali brate 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 9 Apply a pressure of 50 mmHg on the pressure myograph using the Big Ben pressure manom eter When the relative readings at the bottom line of the window appears stable press F3 to proceed 10 Apply a pressure of 125 mmHg on the pressure myograph using the Big Ben pressure ma nometer When the relative readings at the bottom line of the window appears stable press F3 to proceed 11 Apply a pressure of 200 mmHg on the pressure myograph using the Big Ben pressure ma nometer When the relative readings at the bottom line of the window appears stable press F3 to proceed 12 The calibration is now finished The readings in the two bottom lines are the calibration pa rameters stored in the internal memory of the Myo Interface Press ESC to return to the Setup menu 13 Carefully remove the resistance tube shunt and reconnect the tubing to the glass cannulas Open the calibration valve on
20. er Menu Rec Rec Rec Rec Rec Rec Rec Rec Rec Rec Rec Rec l 1 lo 1 hi 2 2 lo 2 hi 3 3 lo 4 4 lo gt 0 200 gt Press 2 E 0 200 gt Force 5 9 z IOO gt Temp E 0 50 F4 Calibrate Pressure Apply 50 mm Hg Push F3 when stable pL SSMA Pp2 3593 F3 Calibrate Pressure Apply 125 mm Hg Push F3 when stable Pal Awe Pas 222 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 18 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL Press 1 gt Calibrate Resovoir Option Info Setup Menu Press Change menu Cal pressure Setup Options Menus 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 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 9 1 1 General Key Functions Je Info Scroll up and down in the menus Being in the top line of a menu use f to change to the previous main menus Push this key in any menu to display all specific F key function
21. esolution Temp probe Weight calibration Analogue output Digital output Voltage Ambient temperature Fluid capacity Gas superfusion ports Valves gt 60 um Manually X Y amp Z settings Glass pipettes Single Max 10 ml 5 ml typical Acid resistant stainless steel With pipes for suction gassing superfusion 50 mN 0 1 mN O 250 mmHg requires external pressure source app 1ATO Manual 0 1 mmHg Built in Ambient 50 C 0 1 C Included Semi automatic 0 1 0 V full scale 12 bit Serial interface RS232 RS485 100 to 240 VAC auto 50 60 Hz 15 30 C 250 ml Built in Safety and reduction Optional accessories FlowMeter range 15 ul min to 1500 ul min Enable pH meter on the interface range pH temp correction 0 14 O C 50 C APPENDIX 2 35 VAS Skejby Science Center Skejbyparken 152 DK 8200 Aarhus N Denmark Tel 45 87 41 11 00 FES ml B LIL OL www dmt dk sales dmt dk support dmt dk DMT Asia Ltd Rm 2402B Great Eagle Centre 23 Harbour Road Wanchai Hong Kong S A R P R China Tel 852 6621 8337 Fax 852 3020 7554 www dmt asia com sales dmt asia com support dmt asia com DMT Asia China office Rm 28C No 8 Dong Fang Road Lu Jia Zui Financial District Shanghai 200120 P R China TES O 21 A25 1S Fax 86 0 21 5877 0063 www dmt asia com sales dmt asia com Support dmt asia com DMT USA Inc 201 East Libe
22. f the blood vessel Left glass cannula Horizontal positioning of the left glass cannula is adjusted by carefully loosening screw D see figure 4 1 on previous page Verti cal positioning of the left glass cannula is adjusted by carefully loosening screw E see figure 4 1 on previous page Longitudinal positioning of the left glass cannula is adjusted by gently loosening of screw F see figure 4 1 on previous page 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 PROTECTION 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 see figure 4 1 on previous page Verti cal positioning of the right glass cannula is adjusted with the micropositioner C see figure 4 1 on previous page Longitudinal positioning of the right glass cannula is adjusted by gently loosening of screw G see figure 4 1 on previous page 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 figure 4 2 and
23. figure 4 3 below Figure 4 2 Three way valve flow settings at P1 Figure 4 3 Three way valve flow settings at P2 CHAPTER4 25 4 1 3 Weight Calibration Procedure The section contains a complete step by step description of how to weight calibrate the force transducer 1 2 Fill the myograph chamber with double distilled water and move the glass cannulas apart Place the calibration bridge balance and weight on the myograph unit allowing it to be pre warmed together with the myo graph unit Turn on the heating in the Main Menu on the Myo Interface 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 figure 4 4 below 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 5 below 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 Figure 4 4 Weight Calibration Set up Figure 4 5 Illustration of how to fit the balance just behind the glass cannula fixation plate 26 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL Go to the Setup Menu on the Myo Interface and choose Force to weight calibrate the force transducer Press F4 to start calibration Make
24. for this connection must be 2 mm a 2 mm tube is not included in the myograph system CHAPTER2 17 CHAPTER 3 THE CONFOCAL PRESSURE MYO INTERFACE The DMT VAS control the Confocal Pressure Myo Interface However the following chapter will go through how to use the Myo Interface without having DMT VAS This chapter is a complete manual for the Confocal Pressure Myo Interface The chapter contains a step by step description of how to navigate in the menus and how to use the special features of the Confocal Pressure Myograph System like the sequencer program 3 1 Myo Interface Manual Figure 3 1 below is an overview of the menus in the Pressure Myo Interface The menus are divided into main menus and sub menus In the following sections all menus are described in detail Myo Interface Model 120CP Version Main Menu 64 4 mm Hg 217 55 3 miming 10 15 mN 520 5 Set Temp 37 0 C Heat on pH 2 KOO Info Change menu Start pres Ctrl Zero force Condensed menu P1 Inlet P2 Outlet Status Flow mm Hg sec Info Change menu Start pres Ctrl Near 12 5mN Far 2 7 mN Act 26 5 Set 37 0 Heat OFF pH 7 00 Setup Options Menus n wr No 7 Comm Menu Serial mode RS485 Myograph No 7 pH Menu Offset buffer LOO Slope buffer 4 00 Temp comp Auto Manual temp e C Figure 3 1 Menus in the 120CP Myo Interface 1 Record
25. he pressure myograph unit as illustrated in figure 4 9 below To prevent the buffer from running into the transducer house the hole is filled with high vacuum grease As a part of daily maintenance it is very important to inspect the greasing of the transducer hole before starting any experiment Insufficient greasing causes damage and malfunction of the force transducer Figure 4 9 Transducer pin hole sealed up with high vacuum grease IMPORTANT DMT RECOMMENDS THAT THE HIGH VACUUM GREASE SEALING UP THE TRANSDUCER HOLE BE CHANGED AT LEAST ONCE A WEEK DMT TAKES NO RESPONSIBILITIES FOR THE USE OF ANY OTHER KINDS OF HIGH VACUUM GREASE THAN THE ONE TO BE PURCHASED FROM DMT DMT TAKES NO RESPONSIBILITIES FOR ANY KIND OF DAMAGE APPLIED TO THE FORCE TRANSDUCERS 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 enable continuous monitoring of the pressure readings from P1 and P2 2 Fill a small syringe with 896 acetic acid and flush both pressure transducers and glass cannulas by connecting the syringe needle to the 3 way valve for the individual pressure transduce
26. inlet pipe Inlet valve Micropositioner for vertical adjustment of glass cannulas Fixation plate for left glass cannula 5 e Outlet connecting pipe t eem e Fixation plate for right glass cannula T PPP Bae miu 9 Silicone tube connecting outlet connecting 9999999996 CHE EEEE E e000000000009000 0000900090400090400090000009000009000900400900090009009 pipe with left glass cannula Left glass cannula HH e Right glass cannula e Myo chamber window Figure 1 2 The Confocal Pressure Myograph chamber close up Silicone Tube connecting perfusion inlet pipe and right glass cannula CHAPTER 1 11 1 2 Confocal Pressure Myo Interface Up and Down Arrow Keys Scroll up and down in the menus If Analogue Recorder Output Connection to external you are at the top of a menu use f to change to the previous 1 data acquisition system optional main menu Myo Interface Display F Keys Have different but specific functions Numeric Keys dependent of the active main menu Enter data or setting values Enter Key Confirm entered numeric values or change between available choices INFO Key Push this key in any menu to display all specific F key functions ESC Key Delete incorrectly entered numeric values In some submenus used to return to main menu Figure 1 3 ConfocalPressure Myo Interface Front 115 230 V 50 60 Hz Automa
27. istilled water 2 Use a swabestick 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 continuing the mechanical cleaning with a swab stick 5 Remove the ethanol solution and wash a few times with double distilled water Incubate the myograph chamber with an 896 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 CHAPTER4 33 APPENDIX 1 FUSE REPLACEMENT The main fuse of the myograph system is placed inside the power inlet on the Myo Interface 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 Red Fuse Block 34 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL APPENDIX 2 SYSTEM SPECIFICATIONS Technical specifications Vessel size Vessel alignment Mounting supports Chamber Chamber volumes Chamber material Chamber cover Force range Force resolution Pressure range Pressure calibration Pressure resolution Heating Temp range Temp r
28. lnerable to applied pressures exceeding 300 mmHg 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 THE PRESSURE APPLIED ON THE PRESSURE TRANSDUCERS WHICH PREVENT THEM FROM BEING DAMAGED THE APPLIED FLUSHING PRESSURE SHOULD NOT EXCEED 150 mmHg This chapter describes how to check the pressure transducers for any kind of damage 4 1 7 1 Simple pressure transducer check 1 If the pressure reading on the Myo Interface appears unstable then first check that the Myo Interface and the Confocal Pressure Myograph Unit are properly connected through the 25 pin grey cable 2 If the pressure reading still appears unstable then perform a new pressure calibration of the pressure transducers as de scribed in 4 1 4 Pressure Calibration Procedure on page 27 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 fthe 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
29. mputer 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 22 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL 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 Temp comp AUTO or MANUAL temperature compensation Manual temp Manual temperature setting The pH submenu contains the settings of the pH calibration 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 between 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 buffers is measured with a thermometer and entered manually in the manual temperature line Press F1 to return to the Setup Menu No 7 pH Menu Offset buffer 7 00 Slope buffer 4 00 Temp comp Auto Manual temp
30. n 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 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 5 Wash the myograph chamber and glass cannulas several times with double distilled water IMPORTANT NOTES BE VERY CAREFUL USING STEP 3 AND 4 REPEATEDLY AS STRONG REAGENTS CAN CAUSE EXTREME DAMAGE TO THE MYO GRAPH 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 SUFFICIENT 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 20 of a 2mM T 1210 Tetrakis 2 pyridylmethyl ethylenediamine solu tion dissolved in double d
31. n 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 read ing T and temperature setting T set Myo Interface Model 120CP Version 3 2 Main Menu Set Temp Heat pH Info Change menu Start pres Ctrl Zero force Condensed menu F4 P1 141 P1 set 140 P2 set 130 pH 7 65 T set 37 CHAPTER 3 19 3 1 4 Pressure Menu The Pressure Menu displays P1 Inlet Inlet pressure setting in mm Hg Pressure Menu P2 Outlet Outlet pressure setting in mm Hg P1 Inlet 90 mm Hg Status Activation of pressure control ON OFF ea Flow Activation of flow control ON OFF Flow ON mm Hg sec Setting of pressure changing gradient The Pressure Menu enables control and adjustment of pressures and flow 1 Inlet and P2 Outlet settings control the inlet and outlet pressure of the perfusion buffer The pressure control is activated or dea
32. n the range of 15 1500 ul min The Flowmeter is eas ily combined with any DMT Pressure Myograph System as illustrated in Figure 2 1 Example of the Complete Confocal Pressure Myograph System Model 120CP on page 14 Use of the FlowMeter 161FM 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 161FM also enables measurement of flow rate as a parameter due to changes in vessel struc ture and function in various physiological 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 a real time flow graph showing flow rate development over time 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 figure 2 2 on previous page to a vacuum pump via a suction bottle and a vacuum valve as illustrated in Figure 2 1 Example of the Complete Confocal Pressure Myograph Sys tem Model 120CP on page 14 The internal diameter of the silicone tube used
33. o 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 controlling the air pressure in the left bottle with bufffer in figure 4 8 on next page Activating P3 serves to prevent a huge pressure to build up in the left bottle with buffer which may damage the mounted vessel when re establishing the flow 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 figure 4 8 on next page 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 figure 4 8 on next page 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 30 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL Air In Max 1 atm Inlet to P1 on Pressure Myograph unit Reduction Valve Manometer 0 5 Bar O
34. oisture does not operate normally or has been dropped 6 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL 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 61326 2 6 2005 EN 61000 3 2 Certified with the safety standards Directive 2006 95 EC EN 61010 1 2001 EN 61010 1 Corr 1 2003 EN 61010 1 Corr 1 2003 EN 61010 2 101
35. on circuit oxygen supply pressure supply and suction device Perfusion outlet Suction pipe for connection to Superfusion outlet Perfusion inlet e vacuum e A e E a J i b i e e n Extra perfusion outlet Superfusion Pipe for connection to Access hole for reagents buffer Extra perfusion inlet inlet oxygen supply temperature or pH probe Figure 2 2 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 9 1 4 Pressure Menu on page 20 To connect the Pressure regulator to the Confocal Pressure Myograph unit use the silicone tube internal diameter 1 mm deliv ered 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 figure 2 1 on previous page NOTE I To fill the tubing and avoid air bubbles mount the Resistance tube shunt as described in 4 1 4 Pressure Calibration Proce dure on page 27 Fill the buffer bottle with 250 ml buffer and run the following flush programme P1 150 mmHg P2 70 mmHg Enter the pressure values in the Pressure menu and press F2 in the main menu to start the flush prog
36. ramme Run the pro gramme 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 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 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 4 1 2 3 way valve adjustments on page 25 16 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL 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 Myograph 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 illustrated in Figure 2 1 Example of the Complete Confocal Pressure Myograph System Model 120CP on page 14 2 3 2 4 Connecting Flowmeter model 161FM optional The DMT Flowmeter model 161FM is optional for flow measurements i
37. rature probe in the buffer solution Sages SIS The third line shows the output from the pH probe as raw data from the A D converter Slope buffer 4 00 Temp auto c DD OE pH output 2 177mV 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 F3 Calibrate pH OFFSET i ff a 22 In the first line is now shown the value of the slope buffer second buffer solution which ee p 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 Special Communication submenu To open the special Communication submenu enter the code 7 in the option line in the Setup Menu No 7 Serial mode RS232 or RS485 serial communication setting Camin Menu i aps Serial mode RS485 Myograph Myograph serial identification A B C or D ec ME Er 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 co
38. rs and glass cannulas using a small piece of silicone tube 32 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL IMPORTANT BE VERY CAREFUL NOT TO APPLY A TOO HIGH PRESSURE WHEN FLUSHING AS THIS MAY DAMAGE THE PRESSURE TRANS DUCERS KEEP AN EYE ON THE PRESSURE TRANSDUCER READINGS ON THE MYO INTERFACE WHEN FLUSHING AND MAKE SURE THAT THE PRESSURE DOES NOT EXCEED 150 mm Hg 3 Flush the pressure transducers and glass cannulas as described in step 2 Repeat the procedure two to three times 4 3 4 Pressure regulator cleaning To clean the buffer circuit port no P2 and 1 on the Pressure regulator follow the procedure described in 4 1 4 Pressure Calibra ion Procedure on page 2 IMPORTANT I BE VERY CAREFUL NOT TO DAMAGE PRESSURE TRANSDUCER P3 WHEN FLUSHING THE BUFFER CIRCUIT THERE ARE NO READINGS FROM P3 VISIBLE ON MYO INTERFACE THEREFORE DMT RECOMMENDS THAT THE PRESSURE MYOGRAPH UNIT ALWAYS BE FLUSHED FIRST THIS WAY IT IS POSSIBLE TO GET A FEELING OF HOW MUCH FLUSHING PRESSURE IS EQUAL TO 150 mmHg IMPORTANT II NEVER FLUSH THE AIR CIRCUITS PORT NO 2 3 AND 4 ON THE PRESSURE REGULATOR WITH ANY KIND OF FLUID 4 3 5 Cleaning pressure myograph chamber DMT STRONGLY RECOMMENDS THAT THE MYOGRAPH CHAMBER AND SURROUNDINGS BE CLEANED AFTER EACH EXPERI MENT After a normal experiment use the following procedure to clean the myograph chamber and supports 1 Fill the myograph chamber to the edge with an 8 acetic acid solutio
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40. s 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 T and keys to adjust the display contrast setting 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 Force Longitudinal force reading in mN Temp Actual temperature probe reading in C Set temp Heating temperature setting in C Heat Actual heating status ON OFF 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 cha nge 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 The Main Menu has the following functions Info F1 F2 F3 F4 Press the Info key and the display shows the F key options i
41. 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 stable Press F3 to pro ceed with calibrating Carefully place the 2 g weight on the pan as illustrated in figure 4 4 on previous page 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 SetupMenu Calibrate Press Resovoir SUC Option 2 0 FA Calibrate Force Apply no Force Push F3 when stable Force 1938 F3 Calibrate Force Place 2 g on the pan Push F3 when stable 8 Press Esc and go to the Main Menu on the Myo Interface The force reading on Force 2245 the Myo Interface should now be very close to 9 81 mN If the force reading is differ ent from 9 81 mN then try to calibrate the fore transducer once again starting with F3 step 3 Calibrate Force DONE Push Esc Force Zero 20315 Force gain 668 3 9 After calibrating the force transducer carefully remove weight balance and calibra tion bridge The Pressure Myograph System is now ready for use longitudinal force measurements 4 1 4 Pressure Calibration Procedure The setup to perform the pressure transducer calibration procedure is illustrated in figure 4 6 below Carefully
42. 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 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 Pressure Myograph System is now ready to be used NOTE A DETAILED GUIDANCE IN HOW TO CONNECT THE CONFOCAL PRESSURE MYOGRAPH SYSTEM FOR AN EXPERIMENTAL SETUP IS DESCRIBED IN DETAIL IN CHAPTER 2 3 BELOW 2 3 Experiment setup 2 3 1 The first weight and pressure calibrations Prior to the shipment of the Confocal Pressure Myograph System 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 Confocal Pressure Myograph System The weight and pressure calibration procedures are described in detail in 4 1 4 Pressure Calibration Procedure on page 27 CHAPTER2 15 2 3 2 External confocal pressure myograph unit connections This section describes how to connect the perfusion circuit the superfusi
43. tic Voltage Selector 9 9 ON OFF Switch 9 Power Connector Jess Ee GEE REGULATOR RS232 pH MYOGRAPH Connection Port for control of Connection Port for Confocal Pressure Regulator Pressure Myograph Unit RS 232 Port for Serial to USB connection to PC Connection Port for pH Meter optional IE ir 12VDCPort Figure 1 4 Pressure Myo Interface Back 12 CONFOCAL PRESSURE MYOGRAPH SYSTEM MODEL 120CP USER MANUAL 1 3 Pressure Regulator Pressure and Perfusion Buffer Inlet and Outlet Pipes l Pressure Transducer Calibration Valve Pressure Valve Activity Indicators Figure 1 5 Pressure Regulator Front Main Pressure Inlet e 115 230 V 50 60 Hz Automatic Voltage Selector Connection Port for the Confocal Pressure Myo Interface e Interface Pressure inlet ON OFF Switch Inlet Pressure Manometer Power Connector Figure 1 6 Pressure Regulator Back CHAPTER 1 13 CHAPTER 2 SETTING UP 2 1 Confocal Pressure Myograph System Model 120CP pH Meter optional Myo Interface Rear Panel Peristaltic Pump optional I Connection cable with i temperature probe PC Data Acquisition and Software RS 232 USB Waste Bottle FlowMeter 161FM I I I I I I I I I I I I I I I I I I I I I 3 I e I optional I I I I I I I I I I I I I I I I I I I I I
44. ufacturer Unplug this apparatus during lightning storms or when unused for long periods of time This apparatus must be grounded Use a three wire grounding type cord similar to the one supplied with the product 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 Voltage 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 m
45. utlet from P2 on Pressure Myograph Unit Relief Valve 0 8 Bar pm A Valve 2 Valve 3 Valve 4 i Air UT 3 No Flow Valve Needle Valve Calibration Valve Buffer Flow Air Flow Figure 4 8 Pressure Regulator Diagram CHAPTER4 31 4 3 Pressure myograph maintenance and cleaning The Confocal Pressure Myograph System Model 120CP is a very delicate and sophisticated 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 clean ing 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 Confocal 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 t

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