Oil Red O Stain for Glycol Methacrylate Sections

Staining Procedures For Plastic Embedded Tissue

Bob Schoonhoven, Laboratory of Molecular Carcinogenesis and Mutagenesis, Dept. of Environmental Sciences and Engineering, University of North Carolina, CB#7400, Chapel Hill, NC 27599


Demonstrate lipids


60% aqueous triethyl phosphate

0.5% Oil Red O solution
0.5g Oil Red O (CI 26125
100.0ml 60% aqueous triethyl phosphate

Filter before use
Celestin Blue
0.5g celestin blue B
100 ml 5% aqueous ferric ammonium sulfate

Boil gently 2-3 minutes; cool to room temperature; filter; and add 12 ml glycerol
Filter before use


Rinse briefly in 60% triethyl phosphate

Stain 5-20 minutes in Oil Red O solution

Rinse 1-2 seconds in 60% triethyl phosphate

Rinse well in distilled water

Counterstain in Celestin Blue 15 minutes

Rinse well in distilled water

Mount in glycerin jelly or other water-soluble mount


lipids: red-orange
nuclei: blue


Feldman, A.T. and Dapson, R.W., "Relative Effectiveness of Various Solvents for Oil Red O," Medical Laboratory Technology, Vol. 31:335-341, 1974.


Energy Beam Sciences manufactures the JB-4 and JB-4A microtomes for sectioning plastic-embedded tissue, and sells GMA kits. Hematoxylin and Eosin stain is a good general stain for many types of tissue.

29b Kripes Road · East Granby, Connecticut 06026-9669
Phone: 800-992-9037 or 860-653-0411 · Fax: 860-653-0422
E-mail: ebs@ebsciences.com