Does it worth to try FACS - (Aug/25/2010 )
Hi everybody
I am working on primary cancer cells. My goal is to find if there is a cooperation between my gene, which is mitotic protein, and the mitotic defect in cancer cells. I have done immunefluorescent experiment. I did not find correlation between the protein level and the mitotic defect. My supervisor wants me to do DNA content analysis experiment using FACS on the same sample set to test the correlation between the gene and the percentage of aneuploid.
She said that FACS can look deeply in the cells and might give different result than IF result. What do you think? Does it worth to try FACS?? Please help me am new PhD student
A good thing about FACS is also that you can measure 10,000 cells in 20 seconds. With IF you can maybe measure 10... So you get more reliable data. Make sure you have good controls!
If your gene is a cytoplasmic protein, it may be difficult to stain your cells for FACS. But not impossible...
rsm
Rsm on Mon Nov 29 09:06:35 2010 said:
A good thing about FACS is also that you can measure 10,000 cells in 20 seconds.
....too slow
Modern cytometers in use (such as LSR-II, Canto II, Aria) are able to measure up to 20-30000 events/sec with appropriate signal CV% (doing that basically everyday in my core facility)