Biotek Synergy LX, User Manual

Page: 124 / 198

100 | Chapter 6 : Instrument Qualification Procedures
l
Sodium Fluorescein Powder (1 mg vial, BioTek PN 98155)
l
A new, clean 96-well solid black microplate, such as Corning Costar #3915, or
equivalent
l
Green filter cube: EX: 485/20, EM: 528/20, Mirror: 510 nm; installed in the reader and
configured in Gen5
Test Solutions
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
any open, unused 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:
l
(Optional, but recommended) Using a 0.45-micron filter, filter 200 mL of deionized
or distilled water.
l
Follow the manufacturer’s instructions on the PBS packaging to create 200 mL,
dissolving the necessary amount of PBS into the filtered water.
l
Stir the solution (preferably using a stir table) until the PBS is completely dissolved.
l
Check the pH; it should be between 7.2 and 7.6 at 25°C.
2. Prepare the sodium fluorescein stock solution:
l
Add 2.0 mL of the buffer solution to the 1 mg Sodium Fluorescein (SF) vial. This
yields a 1.3288 mM stock solution.
l
Ensure that the dye has completely dissolved and is well mixed.
3. Carefully prepare the dilutions. Label each with “SF” and the concentration:
Synergy LX User Manual
1501000N Revision A BioTek Instruments, Inc.