Hello,
I am looking to use Sytox Green as a substitute for propidium iodide (which I can't use with my microscope) in a live cell imaging experiment with T cells killing Daudi Lymphoma cells.
I know that propidium iodide has to be at high concentrations to enter through perforin pores (100uM).
Should I use the same concentration of Sytox green? Any help would be appreciated.
Hello Emily Walsh
SYTOX Green probe may be substituted for PI. SYTOX Green does not display any side effects on cellular viability, cellular proliferation or cell migration when used in concentration upto 1mM. You may use 150nM SYTOX Green. Please see the paper attached below.
Article Real-Time Cytotoxicity Assays
Also, in another article attached below, the investigators explored the interaction of live cells with different fluorescent dyes, over extended periods of time in which cells were continuously cultured in the presence of selected probes: SYTOX Green (SYG, 1–1000 nM), SYTOX Red (SY-R, 1–1000 nM), YO-PRO 1 (1–1000 nM), PO-PRO 1 (1–1000 nM), propidium iodide (PI, 0.1-10 μg/mL), and 7-aminoactinomycin D (7-AAD, 0.1–10 μg/ mL) for up to 5 days. They found that treatment of several different human tumor cell lines in cultures for up to 72 h with the PI, 7-AAD, SYTOX Green (SY-G), SYTOX Red (SYR), TO-PRO, and YO-PRO had no effect on cell viability assessed by the integrity of plasma membrane, cell cycle progression, and rate of proliferation.
Article Chip-Based Dynamic Real-Time Quantification of Drug-Induced ...
Best.
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