[Histonet] Oil Red O for FS on Muscle
macveigh <@t> usc.edu
Thu Jan 27 15:09:20 CST 2011
Order the stain from Electron Microscopy Sciences - cat # 26503-02. It
comes as 250 ml solution and is a lot less expensive than some other
sources. I guess you can probably get it through VWR.
Mix 3 parts of this stock with 2 parts dist. Water and let it stand for 10
Filter through coarse filter paper. I have even used paper towel with no
problem. Anything finer will take forever to filter.
Fix the sections with regular 10% NBF for few min. 3-5
Rinse slides in dist. Water few times
Working Oil -red-O for 10 min. I use the plastic containers for shipping
slides, because the stain will color the plastic and I can easily and
cheaply replace those, but you can wash them and reuse them multiple times.
70% alcohol - quick rinse to clear background
Dist. Water - 5 min
Mayers Hematoxylin 5 min
Tap water 10 min
Dist. water rinse or up to 1 min
Mount with Aqua Mount
Never had any problems. Normal muscle will have very, very tiny droplets. We
used it on rodent muscle, but it should work the same for human.
USC Keck School of Medicine
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