I would like to try to either coat plates with fibronectin or fix the cells in acetone ahead of time to save time during my IF assay. Can I coat the glass in fibronectin and let dry to use at a later date (or store in PBS?)? Or, can I fix cells with acetone and store for later use? Any details storage temperature, protocols, or advice are appreciated! Thank you!
Hi Erica Briggs . I know that you can coat glass slides with fibronectin (10 ug/ml) in PBS for an hour at RT, wash with PBS, shake off the liquid and store them dry at -80oC (not -20oC!!!) in a closed container for over a year and they are still good.
Hi Erica
With respect to fixation of cells with Acetone and their storage. We routinely fix cells on coverslips (coated or uncoated) in pure acetone for 5-7 min at -20 C. Samples are rinsed thrice in ice cold PBS and for short term storage (1 week to 1 month) are stored in 0.1% PBS-Azide solution at 4C. For medium term storage (2-6 months) coverslips are mounted on a semi-solid mounting media (fluoromount G) on a glass slide (allowed to dry), followed by packaging these slides in plastic bags with dessicating agents (silica beads in pouches) and sealed for storage at 4C. When needed for future use, we hydrate the coverslips till the mounting medium comes loose and coverslip can be lifted with a fine tip forceps.
Hope this helps
Best
Aparajita
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