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Oil Red O won't stain cells


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#1 Cuse Grad

Cuse Grad

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Posted 14 July 2012 - 08:22 AM

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 cells

at 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!






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