29
1. Mount and normalize the vessels as described
in the former chapters.
2. Perform a standard start and check the
vessel segment for endothelium function, as
described in chapter 3.4 and 3.5.
3. Add noradrenaline to obtain a response
around 60% of maximum (determined from
the previous noradrenaline concentration-
response curve). When the contractile
response is stable, add increasing
concentrations of acetylcholine to the
chamber, using the table below as a guideline.
Wait for a stable contractile response or a
standard time such as two minutes between
each application.
3.7.2 PROTOCOL
Prepare the following stock solutions:
Acetylcholine:
10
-4
, 10
-3
, 10
-2
M
Noradrenaline:
10
-2
M
[ACh] in chamber (µM)*
Volume of stock solution to add to chamber
0.1
5 μL of 10
-4
M
0.3
1 μL of 10
-3
M
0.5
1 μL of 10
-3
M
1
2.5 μL of 10
-3
M
1.3
1.5 μL of 10
-3
M
1.5
1 μL of 10
-3
M
3
7.5 μL of 10
-3
M
5
1 μL of 10
-2
M
10
2.5 μL of 10
-2
M
*In calculating the [ACh] in the Wire Myograph chamber, the applied volume of ACh is ignored.
Содержание 630MA
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