I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using glycerol as the medium and placing coverslips on the slides and observing them but the coverslips are not stable on the slides causing detachment of the cells. Should i try fixing my cells?? will it affect the proteins in the cells??
You should probably perform a light fixation to prevent degradation of the cells in the glycerol. There is always a possibility that fixation might interfere with your proteins, but in my experience fluorescent proteins still perform well after fixation.
If you're having issues with the coverslips falling, I'd suggest using a clear nail polish to seal the edges and better adhere the coverslip to the glass slide.
John Hardy Lockhart thank you for your response. I will try what you suggested
John Hardy Lockhart . Is 4% paraformaldehyde considered a light fixation method??
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