Preparation of acid-washed glass coverslips for immunofluorescence microscopy
Jenna Ekstrom
Published: 2024-05-17 DOI: 10.17504/protocols.io.6qpvr847blmk/v1
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Abstract
Rizzardi Lab (Adapted from Schwer Lab- UCSF)
Steps
1.
Add coverslips to 10cm culture dish
2.
Add ~20 mL of 1 N HCl; swirl on orbital shaker for 2 h at RT.Room temperature
3.
Remove acid, rinse with 2 x 25 mL PBS until pH is at least 6.0.
4.
Rinse 1 x 25 mL water.
5.
Add ~20 mL of 70% EtOH; swirl on orbital shaker for 10 min, RT.Room temperature
6.
Remove ethanol, transfer coverslips to Whatman paper. Let dry in TC hood.
7.
Sterilize coverslips before use
7.1.
Option 1:
Transfer to glass beaker, cover with aluminum foil and autoclave (dry cycle, 30 min sterilization time)
7.2.
Option 2:
Transfer to new sterile 10cm culture dish and turn on UV. Leave in hood until ready for use.
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