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TISSUE BATH

MYOGRAPH

SYSTEM

- 720MO

- USER GUIDE

USER GUIDE

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Summary of Contents for 720MO

Page 1: ...1 TISSUE BATH MYOGRAPH SYSTEM 720MO USER GUIDE USER GUIDE ...

Page 2: ...TISSUE BATH 720MO USER GUIDE ...

Page 3: ...tart 8 3 3 1 Principles of the standard start procedure 8 3 4 Endothelium function 9 3 4 1 Principles of checking endothelium function 9 3 5 In vitro experiment 1 Noradrenaline contractile response 9 3 5 1 Background 9 3 6 In vitro experiment 2 Acetylcholine relaxation curve 10 3 6 1 Background 10 3 5 2 Protocol 10 3 6 2 Protocol 11 Chapter 4 CleAning and maintenance 12 4 1 Cleaning the Tissue Bat...

Page 4: ...R 1 CHAPTER 1 TISSUE BATH OVERVIEW Micrometer Pin support connected to micrometer Force transducer pin Pin support connected to force transducer Figure 1 1 Tissue Bath with close up of chamber Transducer house ...

Page 5: ...L WHEN CHANGING OR AD JUSTING THE MOUNTING SUPPORTS 2 1 1 Changing the mounting supports figure 2 1 1 Use the micrometer to separate the supports as far apart as possible 2 Use the small screwdriver provided to gently loosen screw D on the support attached on the transducer side Screw D is the screw on the support closest to the transducer 3 Gently pull the support away from the transducer pin 4 L...

Page 6: ...crew C to align the transducer side tissue holding pin horizontally 4 Loosen screws D and E to align the heights of the tissue holding pins vertically 2 2 Calibration of the force transducer As a part of the general maintenance of the Tissue Bath DMT recommends that the Tissue Bath is force calibrated at least once a month The Tissue Bath should also be force calibrated every time the Multi Interf...

Page 7: ...ollowing thickness L shaped mounting pins Diameter 200µm 250µm 300µm and 400µm 1 Fill the chamber with a given buffer 5 7ml see appendix 1 for Buffer recipes DO NOT FILL THE CHAMBER TO THE EDGE OF THE CHAMBER 2 Make sure that the tissue holding pins are properly aligned 3 Use the micrometer to get the pins close together as possible without touching each other 4 Slide your tissue ring preparation ...

Page 8: ...ension T mN mm respectively Plotting wall tension against internal circumference reveals an exponential curve and by applying the isobar curve corresponding to 100 mmHg IC100 is calculated from the point of intersection using the Laplace relation figure 3 9 IC1 is calculated from IC100 by multiplying a factor giving an internal circumference at which the active force production as well as the sens...

Page 9: ...relaxation of the vascular smooth muscle cells If the endothelium is undamaged by the dissection and mounting procedures then a substantial relaxation will occur With complete removal or damaged endothelium a partial relaxa tion or no relaxation to acetylcholine is observed It is important to note that the amount of NO or EDRF in a vessel is often dependent upon its size In certain vessels endothe...

Page 10: ...sence of cocaine to block the noradrenaline reuptake To determine the sensitivity to noradrenaline the vessel segment is exposed to increasing concentrations of noradrenaline Each concentration is applied until a steady response has been reached and then the next concentration is applied When the vessel segment is fully contracted or does not response more upon increasing the noradrenaline concent...

Page 11: ...onse curve as described in the previous section The vessel segment is exposed to the noradrenaline concentration and when the response has stabilised increasing concentra tions of acetylcholine are added to relax the vessel Each concentration is applied until a steady response has been reached and then the next concentration is applied When the vessel segment is either fully relaxed or does not re...

Page 12: ...aining in the chamber or on the tubing using absorbent paper IMPORTANT NOTES BE VERY CAREFUL USING HCL OR HNO3 BECAUSE THESE ACIDS MAY CAUSE EXTREME DAMAGE TO THE STAINLESS STEEL CHAMBERS AND SUPPORTS DO NOT USE BLEACH TO CLEAN THE CHAMBERS REPEATED USE OF CHLORINATED SOLUTIONS SUCH AS BLEACH AND HCL WILL CAUSE DAMAGE TO THE STAINLESS STEEL PARTS OF YOUR TISSUE BATH AVOID USING THEM IF AT ALL POSS...

Page 13: ...ery vulnerable to applied forces exceeding 1 Newton 100 grams or fluid running into the transducer compartment due to insufficient greasing of the transducer pinhole If the force readings on the Multi Interface appear unstable or noisy then first check that the chambers are connected properly to the Multi Interface and that the chambers are plugged all the way into the Multi Interface If the force...

Page 14: ...around the bottom of the transducer housing to seal the transducer housing when set back in place see figure 4 3 6 Carefully realign the transducer housing with the new transducer on the Tissue Bath and reinsert the Allen screws through the bottom of the Tissue Bath 7 Tighten the screws and place some vacuum grease around the transducer pin that protrudes from the transducer housing see figure 4 3...

Page 15: ... linear slides under the black covers for grease at least once a week In case of insufficient lubrication grease the slides with the Grease for Linear Slides included with your system See figure 4 4 Figure 4 4 The areas where linear slide grease may be applied for smooth micropositioner movement ...

Page 16: ... while being constantly stirred For example if 1 litre of PSS is to be made then dissolve all the chemicals in 800mL of double distilled H2 O 3 Add the appropriate volume of 1 0M CaCl2 for the total volume of PSS being made for example 1 6mL of 1 0M CaCl2 for 1 litre of buffer Continue to stir the PSS while the CaCl2 is being added 4 Bring the solution up to the final volume with double distilled ...

Page 17: ... 8 73 17 46 KCl 74 557 60 2 24 4 47 8 95 17 89 KH2 PO4 136 09 1 18 0 08 0 16 0 32 0 64 MgSO4 7H2 O 246 498 1 17 0 145 0 29 0 58 1 16 NaHCO3 84 01 14 9 0 625 1 25 2 50 5 00 Glucose 180 16 5 5 0 5 1 00 2 00 4 00 EDTA 380 0 026 0 005 0 01 0 02 0 04 CaCl2 110 99 1 6 0 8mL 1 6mL 3 2mL 6 4mL 1 0 M solution 1 Make a 1 0M solution of CaCl2 110 99 in double distilled H2 O Filter sterilize the calcium solut...

Page 18: ...are just separated and is given by IC0 2 d where d is the wire diameter For 40 µm wires IC0 205 6 µm Using the Laplace relation the effective pressure Pi is calculated for each pair of readings The effective pressure is an estimate of the internal pressure which is necessary to extend the vessel to the measured internal circumference Pi Ti ICi 2 The stepwise distension is continued until the calcu...

Page 19: ...H SYSTEM 720MO USER GUIDE The normalized internal lumen diameter is then calculated by IC1 d1 The micrometer reading X1 at which the internal circumference of the normalized vessel is set to is calculated by IC1 IC0 2 X1 X0 ...

Page 20: ...pped at a point beyond 10 on the sleeve indicating 10000 µm 10 mm 2 Note that there is no mark completely visible between the 10 mm mark and the thimble 3 Read the value on the thimble corresponding to the intersection with the horizontal line on the sleeve A Reading on sleeve B No additional mark visible C Thimble reading Total reading Example 2 1 Note that the thimble has stopped at a point beyo...

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