Biotek Cytation 5, Instructions For Use Manual

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70 | Instrument Testing
Test Solutions
Corners/Sensitivity/Linearity Tests
If using BioTek’s sodium fluorescein powder (PN 98155), be sure to hold the
vial upright and open it carefully; the material may be concentrated at the top.
If a centrifuge is available, spin down the tube before opening.
When diluting the sodium fluorescein powder in buffer, it takes time for the
powder to completely dissolve. Allow the solution to dissolve for five minutes,
with intermittent vortexing, before preparing the titration dyes.
Wrap the vial containing the stock solution in foil to prevent exposure to light.
Discard unused solution after seven days. Discard any open, unused buffer
solution after seven days.
1. The Sodium Borate solution does not require further preparation; proceed to
step 2. If you are using PBS, prepare the solution:
• (Optional, but recommended) Using a 0.45-micron filter, filter 200 mL of
deionized or distilled water.
•
Follow the manufacturer’s instructions on the PBS packaging to create 200
mL, dissolving the necessary amount of PBS into the filtered water.
• Stir the solution (preferably using a stir table) until the PBS is completely
dissolved.
• Check the pH; it should be between 7.2 and 7.6 at 25°C.
2. Prepare the sodium fluorescein stock solution:
•
Add 2.0 mL of the buffer solution to the 1 mg Sodium Fluorescein (SF)
vial. This yields a 1.3288 mM stock solution.
•
Ensure that the dye has completely dissolved and is well mixed.
3. Carefully prepare the dilutions. Label each with “SF” and the concentration:
Mix This SF Solution:
With
Buffer:
To
Make:
0.53 mL of 1.3288 mM stock
solution
13.47 mL 50.2 μM
110 µL of 50.2 μM SF 13.89 mL 400 nM
3.5 mL of 400 nM SF 10.5 mL 100 nM
0.46 mL of 100 nM SF 13.54 mL 3.3 nM Corners Test
4.24 mL of 3.3 nM SF 9.76 mL 1 nM
Sensitivity/Linearity
Tests
BioTek Instruments, Inc.