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Hello,
I've been staining fixed, adult stem cells with oil red o. A few of my runs didn't go too well- a lot of excess staining
on well walls prior to eluting. Upon attempting to stain
a second time, my cells didn't stain at all. Any help would be greatly appreciated! Here's a bit more about my protocol:
Stock solution: 0.5% ORO in 100% Isopropanol, Working solution: 3:2 solution of ORO:
H2O , made 10 min before use
1) Cells fixed with formalin
2) Incubate cellsat RT in 60% Isopropanol, 5 min
3) Add filtered working solution, incubateat RT for 15 min
4) Wash wells carefully with 1xPBS
5) Elute in 100% Isopropanol, 0.5-1 hr
6) Remove elution
I had been adding PBS after the elution, also, before attempting my second staining
attempt.
Thanks!
