I am currently doing senesecence-related beta-gal staining. The problem is I have to spend a lot of time to count the cells, which was indeed countless.
Does any one have an idea about how many cell counts are considered "enough"? How many different scopes should be taken in consideration of repitition?
You need to count at least 1000 TOTAL cells irrespective of number of fields to get a true assessment of percentage positivity. The fraction of SA-beta-gal positive cells within this 1000 will vary but this method will give a good statistically significant value as used in many labs who routinely score positive cells for a wide range of markers!
Dear Siyuan,
Did you ever figure out a good method for counting beta-gal cells? I have started using DAPI to count total cells but I'm still searching for an automated or semi-automated method to set an appropriate threshold for the blue signal and count the number of SAbeta-gal cells.
Also - I am working with 12-well plate cultures and sometimes I do not have 1000 cells in each well. Have you tried using the 1000 cell rule to determine degree of senescence between samples? Did it work well?
Best,
Sindhu
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