I have plated some glioblastoma cells in a 24 well/plate, and am looking to see the impact that adding DHA (the fatty acid) to these cells has. From my reading I expect there to be an accumulation of lipid droplets, followed by an increase in cell death (ferroptosis) as the excess lipids undergo peroxidation. I was planning on staining the cells after 72 hours, however by this point surely the cells will be dead. Most papers stain after 24 hours. I only have one plate seeded.
Should I stain after 24 hours, and if so, can I continue to image the cells at 48 and 72 to see if their survival rate after each day?
Oil red O is used on fixed cells, but there are live cell-compatible lipid droplet stains. Lipid droplets have a polar lipid shell around a neutral lipid core. Oil red O stains the neutral core. Live cell fluorescent dyes are available for available for neutral and polar lipids And you could stain your cells at 24 hours.
https://biotium.com/product/lipidspot-488-lipid-droplet-stain-1000x/
i have used the DyRect live cell lipid stains from MarkerGene, now part of Abcam.
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