Jacobs:Protocol Oil Red O Staining: Difference between revisions

Latest revision as of 19:04, 16 June 2008

Protocol for Oil Red O Staining. Useful for identifying adipocytes.

Citrate Working Solution (store in 4C for long term use)
- 2mL citrate concentrate solution
- 8mL deionized water
Oil Red O Stock Solution
- 300mg Oil Red O
- 100mL 99% isopropanol
Staining Solution
- 6 parts of stock Oil Red O
- 4 parts of distilled water
- Allow to stand for 10min
- Filter this solution with Whitman paper
- Working solution is stable for 1-2 hr

@@ Line 39: / Line 39: @@
 ==Procedure==
+# Warm citrate working solution
+# Prepare fixative solution: 2 parts citrate working solution + 3 parts acetone
+# Remove media
+# Wash cells 2x with PBS
+# Fix with 1ml fixative solution for 30sec
+# Wash cells 1x with distilled water for 45sec.  Do not allow plates to dry.
+# Stain in 1ml staining solution for 6-15min
+# Clear background using 1ml 60% isopropanol
+# Wash well in distilled water
+# Counter stain with 1ml hematoxylin for 5min
+# Rinse with distilled water
+* Lipid: Red
+* Nuclei: Blue
-==Notes==
-Used in Stanford for Tissue Engineering Lab Course (ME385B)and 2007 Winter TC workshop.
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-==References==
-<!--'''Relevant papers and books'''
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 ==Contact==
@@ Line 62: / Line 59: @@
-or instead, [[Talk:{{PAGENAME}}|discuss this protocol]].
+[[Talk:{{PAGENAME}}|discuss this protocol]].
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