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using automated fluorescence microscope rather than flow cytometry

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#1 TTT



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Posted 31 May 2010 - 01:49 AM

Hi, we dont have a FACS machine in the lab so we cant do flow cytometry, what we do have is automated fluorescence microscope (ImageXpress Micro). I am going to do an apoptosis assay using Propidium iodide, Hoescht and YO-PRO-1 dyes. However the protocol i got from the company are for flow cytometry use, Im just not sure how i can adapt a protocol for flow cytometry to fluorescent microscope, i guess with ImageXpress, my cells are all attached to the plate whereas in the flow cytometry protocol, the cells are in solutions. So I have questions like do I remove the dyes before imaging, do i fix cells or not etc, has anyone done apoptosis study with fluorescent microcope rather than flow cytometry and has a good protocol for me to adapt, thanks.

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