My cells are fixed on a coverslip and are stained with DAPI, Cytokeratin and CD45. What steps should I take after staining the cells?
I do not think you need a special protocol. Use 10-20ul of antifade solution depending upon the slide size where cells are and cover with a cover slip. Be sure to avoid air bubbles. Keep slides in dark area on a a horizontal slide organizer. You can keep in refrigerator for several days or you can see immediately under microscope. You may or may not use nail polish around the rim of coverslip to avoid slippage of material outside. That is subjective. Keep antifade solution wrapped and at 4C
I do not think you need a special protocol. Use 10-20ul of antifade solution depending upon the slide size where cells are and cover with a cover slip. Be sure to avoid air bubbles. Keep slides in dark area on a a horizontal slide organizer. You can keep in refrigerator for several days or you can see immediately under microscope. You may or may not use nail polish around the rim of coverslip to avoid slippage of material outside. That is subjective. Keep antifade solution wrapped and at 4C
The step after adding the antibodies and washing is the antifaid - mounting step. You should be careful with this step because you need a small amount of mounting. Once you put the mounting on the slide put the cover slip slowly. Make sure no bubble are formed. Leave them in the dark at room temperature or in the fridge for 20-30 minutes. Then you can do the sealing step.
Both the above methods are correct, however, after mounting your slide with Anti-fade, you must keep it at RT in dark till the anti-fade gels up. This may require an overnight step at RT. The gelling of anti-fade does not happen very well at 4C. Depending on your type of microscopy, the gelling of anti-fade may/may not have an effect on images. Once the gel is set, you can store your slides for a few months in 4C.
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