Related Manuals for DMT 630MA
Summary of Contents for DMT 630MA
- Page 1 MYOGRAPH SYSTEM - 630MA USER GUIDE VOL. 3.0 Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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Page 2: Table Of Contents
APPENDIX 1 - BUFFER RECIPES PHYSIOLOGICAL SALINE SOLUTION (PSS) HIGH POTASSIUM PHYSIOLOGICAL SALINE SOLUTION (KPSS) APPENDIX 2 - NORMALIZATION THEORY MATHEMATICAL CALCULATIONS APPENDIX 3 - READING A MILLIMETRE MICROMETER Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... -
Page 3: Chapter 1 - Wire Myograph Overview
CHAPTER 1 - WIRE MYOGRAPH OVERVIEW Figure 1.1 Close up of Multi Myograph unit and myograph jaws Temperature probe Funnel For drug application Figure 1.2 Chamber cover Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... -
Page 4: Chapter 2 - The Multi Wire Myograph Unit
3. Gently pull the support away from the transducer pin. 4. Loosen screw B on the micrometer side with the appropriate fitting Allen key. 5. Pull the support away. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 5 4. Loosen screw D to move the jaw to the right or left on the transducer side. Figure 2.1 Illustration of the screws for changing supports and coarse adjustment of the jaws Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 6 Figure 2.2 Fine adjustments of the jaws in the Wire Myograph chamber Figure 2.3 - Illustrations of properly aligned jaws (depicted on the far left) and incorrectly aligned jaws (depicted in the middle and far right). Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 7 Figure 2.4 - Fine adjustments of the pins in the Myograph chamber Figure 2.5 - Illustrations of properly aligned pins (depicted on the far left) and incorrectly aligned pins (depicted in the middle and far right). Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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Page 8: Calibration Of The Force Transducer
2.2 CALIBRATION OF THE FORCE TRANSDUCER As a part of the general maintenance of the Wire Myograph, DMT recommends that the Wire Myograph is force calibrated at least once a month. The Wire Myograph should also be force calibrated every time the interface has been moved. Although lab benches are all supposedly perfectly horizontal, small differences in lab bench pitch can affect the calibration of the system. -
Page 9: Chapter 3 - Experimental Set-Up
(figure 3.1 B-C). • Fill the Wire Myograph chamber with PSS (at room temperature). See appendix 1 for example of buffer recipes. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 10 (figure 3.2 A-C). Be careful not to stretch the vessel segment if the end of the wire catches the vessel wall. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 11 (figure 3.4 B). The excessive vessel segment is finally dissected free and removed from the wire (figure3.4 C). Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 12 (connected to the transducer). Secure the near end of the wire in a clockwise direction under the far fixing screw on the right-hand jaw (figure 3.6 B). Figure 3.6A and B Mounting step 6 Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 13 Use a dissection microscope at this step while moving the jaws together using the manual micropositioner until the two wires are as close to each other as possible but without touching. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 14 This is a side view of the two wires on the two. • The artery is now mounted correctly and can be moved onto the 630MA interface with the heat turned on. When the buffer in the chamber has reached 37 C, replace the buffer in the chamber with fresh well aerated warm buffer.
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Page 15: Mounting Protocol For Larger Arteries
For mounting of larger arteries with an internal diameter larger than 500µm and up to 11mm replace the mounting Jaws with Pins (see chapter 2.1). With the 630MA system a box with 4 sets of 200µm pins is delivered. For large arteries pins with 250µm, 300µm or 400µm pins can be purchased from DMT. - Page 16 Take the isolated artery/tissue section and slide it over the two pins as shown in the figures below. The artery is now mounted correctly and the heat of the 630MA interface can be turned ON. When the buffer in the chamber has reached 37C, replace the buffer in the chamber with fresh well gassed warm buffer. Wait 5 min and start the Automated Normalization as described in Chapter 3.3.5.
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Page 17: Normalization
) that a fully relaxed segment would have at a specified transmural pressure. For small rat arteries the target transmural pressure is typically 100mmHg = 13.3kPa. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 18 1. Zero the force of the Chamber going to be SELECT CHAMBER 2: normalized automatically. SELECT SELECT CHAMBER 3: SELECT 2. Go into the Norm. Set-up menu. SELECT CHAMBER 4: SELECT ENTER Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 19 Pressure is 13.3kPa and the Norm Factor is 0.9 but for mouse mesenteric arteries the Norm Pressure is 13.3kPa and the Norm Factor 1.1. To find the Norm factor for your specific artery and specie please read DMT Normalization Guide. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 20 8. Select the appropriate Chamber MOUNT ON CHAMBER NO. SELECT CHAMBER 1 SELECT SELECT CHAMBER 2 SELECT SELECT CHAMBER 3 SELECT SELECT CHAMBER 4 SELECT ENTER Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 21 If the normalization fails it can always be repeated with new values for the Norm Force. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 22 Step no.: 2 Time: 0 Xo: +0.00 um Yo: +48.13 mN Force: +53.14 mN Not enough steps to normalize. GO BACK and check parameter. BACK Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 23 Mount chamber 4 and start the automated normalization when chamber 3 has finished 20. All chambers are now mounted, and the Standard start procedure can be started. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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Page 24: Standard Start
37 and aerated with a mixture of 95% O and 5% CO before use. Instructions °C for making the necessary solutions are described in appendix 1. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... -
Page 25: Endothelium Function
(EDCF). Therefore, it is important to check the existing literature in order to determine the expected response in your particular vessel with the given concentration of agonist. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... -
Page 26: In Vitro Experiment 1: Noradrenaline Contractile Response
When the vessel segment is fully contracted or does not respon d more upon increasing the noradrenaline concentration, the experiment is ended. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 27 1μL of 10 7.5μL of 10 1μL of 10 2.5μL of 10 *In calculating the [NA] in the Wire Myograph chamber, the applied volume of noradrenaline is ignored. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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Page 28: In Vitro Experiment 2: Acetylcholine Relaxation Curve
When the vessel segment is either fully relaxed or does not relax more upon increasing the acetylcholine concentration, the experiment is ended. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 29 1μL of 10 7.5μL of 10 1μL of 10 2.5μL of 10 *In calculating the [ACh] in the Wire Myograph chamber, the applied volume of ACh is ignored. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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Page 30: Chapter 4 - Cleaning And Maintenance
Turn off the vacuum and gas supply. Remove any water or buffer remaining in the chamber or on the tubing using absorbent paper. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 31 12. Wash the chamber and supports several times with double distilled water. 13. Dry the surfaces using absorbent paper (i.e. Kim-Wipes) or cotton-tip applicators. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 32 PROPER CLEANING OF ALL SUPPORT SURFACES. NEVER SOAK THE SUPPORTS IN ANYTHING STRONGER THAN 8% ACETIC ACID FOR EXTENDED PERIODS OF TIME (I.E. SEVERAL HOURS OR OVERNIGHT). Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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Page 33: Maintenance Of The Force Transducer
Figure 4.1 Close-up of transducer pin from outside. The arrow indicates the place that the grease needs to be applied to prevent water and buffer from damaging the transducer. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 34 3.5.1 in 630MA Myograph System User Manual. Electrical noise from the surroundings can be removed by grounding of the system using the ground connection on the back side of the 630MA system. During the new calibration, monitor the relative force reading values in the Force Calibration sub -menu on the Multi Interface the normal operating values for the force transducer during calibration should be between 3000 and 3500.
- Page 35 IMPORTANT: CALIBRATE THE NEW FORCE TRANSDUCER BEFORE PERFORMING A NEW EXPERIMENT. Figure 4.3 - The 8 screws that secure the transducer house to the chamber Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 36 Figure 4.4 Inside the transducer housing and close-up of transducer pin. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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Page 37: Maintenance Of The Linear Slides
“Grease for Linear Slides” included with your system. (See figure 4.5 below). Figure 4.5 The areas where linear slide grease may be applied for smooth micropositioner movement Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... -
Page 38: Appendix 1 - Buffer Recipes
This takes about 15 minutes at room temperature. Aerate the solution with carbogen (95% O 2 + 5% CO 2 ) for about 20 minutes. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 39 25x Concentrated PSS (Stock solution): Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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Page 40: High Potassium Physiological Saline Solution (Kpss)
HIGH POTASSIUM PHYSIOLOGICAL SALINE SOLUTION (KPSS) 1x 60mM KPSS: Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... -
Page 41: Appendix 2 - Normalization Theory
Using the Laplace relation, the effective pressure P i 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. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 42 The normalized internal (lumen) diameter is then calculated by: The micrometer reading X1 at which the internal circumference of the normalized vessel is set to is calculated by: Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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Page 43: Appendix 3 - Reading A Millimetre Micrometer
0.5mm. Each division on the thimble scale is 10µm. If the thimble scale falls between two lines, then a number between 0 and 10 µm must be app roximated. Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk... - Page 44 3. Read the value on the thimble corresponding to the intersection with the horizontal line on the sleeve. Reading on sleeve: 16000µm 500m One additional mark visible: 280µm Thimble reading: Figure A2.3 Example 2: reading = 16780µm 16780µm Total reading: Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
- Page 45 Danish Myo Technology A/S | Tel.: +45 87411100 | Email: support@dmt.dk | Web: www.dmt.dk...
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