DMT 720MO, Руководство пользователя

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17 TISSUE BATH MYOGRAPH SYSTEM - 720MO - USER GUIDE
1. Make a 1.0M solution of CaCl
2
(110.99) in double-distilled H
2
O. Filter-sterilize the calcium solution through a 0.22 μm filter.
The sterilized solution can be stored in the refrigerator for up to 3 months.
2. Dissolve all the chemicals except the CaCl
2
in approximately 80% of the desired final volume of double distilled H
2
O 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 H
2
O.
3. Add the appropriate volume of 1.0M CaCl
2
for the total volume of PSS being made (for example, 1.6mL of 1.0M CaCl
2
for 1
litre of buffer). Continue to stir the PSS while the CaCl
2
is being added.
4. Bring the solution up to the final volume with double-distilled H
2
O. Continue to stir the solution until the EDTA is fully dis-
solved. This takes about 15 minutes at room temperature.
Before use:
5. Dilute the 25 x PSS stock solution 1:25 using double distilled H
2
O.
6. Add:
1.091 g/L Glucose
2.100 g/L NaHCO
3
7. Aerate the solution with carbogen (95%O
2
+ 5%CO
2
) for at least 20 minutes. If necessary wait further for the pH of the buffer
to reach pH 7.4.
High potassium Physiological Saline Solution (KPSS)
1x 60mM KPSS:
Chemical Mol.Wt mM g/0.5L g/L g/2L g/4L
NaCl (58.45) 74.7 2.18 4.37 8.73 17.46
KCl (74.557) 60 2.24 4.47 8.95 17.89
KH
2
PO
4
(136.09) 1.18 0.08 0.16 0.32 0.64
MgSO
4
7H
2
O (246.498) 1.17 0.145 0.29 0.58 1.16
NaHCO
3
(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
CaCl
2
(110.99) 1.6 0.8mL 1.6mL 3.2mL 6.4mL
(1.0 M solution)
1. Make a 1.0M solution of CaCl
2
(110.99) in double-distilled H
2
O. Filter-sterilize the calcium solution through a 0.22 μm filter.
The sterilized solution can be stored in the refrigerator for up to 3 months.
2. Dissolve all the chemicals except the CaCl
2
in approximately 80% of the desired final volume of double distilled H
2
O 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 H
2
O.
3. Add the appropriate volume of 1.0M CaCl
2
for the total volume of PSS being made (for example, 1.6mL of 1.0M CaCl
2
for 1
litre of buffer). Continue to stir the PSS while the CaCl
2
is being added.
4. Bring the solution up to the final volume with double-distilled H
2
O. Continue to stir the solution until the EDTA is fully dis-
solved. This takes about 15 minutes at room temperature.
5. Aerate the solution with carbogen (95% O
2
+ 5% CO
2
) for about 20 minutes.
Calcium-free physiological saline solution (Ca-free PSS)
To make 1 L of Ca-free PSS:
Use the recipe for regular PSS but omit the CaCl
2
and add EGTA (to buffer the residual Ca
2+
in solution). The concentration of EGTA
in the PSS should be 1-5 mM to ensure sufficient buffering.