Hi, everybody. Currently, I am working with the cell cultures (HEK293 cell line). I'm looking for the most common (preferably most affordable dyes, too) that will efficiently label proliferative or dying cells for either flow cytometry or fluorescent microscopy, or perhaps both. Thx.
For labelling dead cell you can use Ethidium Monoazide, it's a cheap dye and works well with eukaryote cells. As it's photoactivable you can fix the staining in one specific time interval.
the classic one is trypan blue exclusion test by standard transmition light microscopy
for cytometry you can use DIOC6, you can use it too for fluorescent microscopy but better for cytometry
Anexin combined with DIOC6 can detect cell death in cytometry
combined with DIOC and anexin, you can use propidioum iodide, with will detect dead cells
JC-1 can be used for cell death in fluorescent microscopy
hope these pointers help
best
JP
CFDA-SE to perform a CFSE dilution assay for cell proliferation is standard flow cytometry protocol. Not very expensive.
7AAD for cell death assay in flow cytometry.
Hi,
I suggest you to go for nuclear staining using Propidium iodide for identifying dead cells and DAPI for live and fixed cells. Both the dyes can be used for flow cytometry and fluorescence microscopy. PI with hoechst stain together gives you more differential observations to identify live and dead cells by flow cytometry and fluorescence microscopy.
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