9/17/97

Dorothy and fellow microscopists,



I have a procedure for Oil Red O staining of GMA sections which was sent to

me by Bob Schoonhoven. I haven't tried it myself.



Stain: Oil Red O

Indication: Demonstrate lipids

Solutions:

1) 60% aqueous triethyl phosphate

2) 0.5% Oil Red O solution

0.5g Oil Red O (CI 26125)

100 ml 60% aqueous triethyl phosphate

Filter before use

3) 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

Procedure:

1) Rinse briefly in 60% triethyl phosphate

2) Stain 5-20 minutes in Oil Red O solution

3) Rinse 1-2 seconds in 60% triethyl phosphate

4) Rinse well in distilled water

5) Counterstain in Celestin Blue 15 minutes

6) Rinse well in distilled water

7) Mount in glycerin jelly or other water-soluble mount

Results:

lipids: red-orange

nuclei: blue

Reference:

Feldman, A.T. and Dapson, R.W., "Relative Effectiveness of Various Solvents

for Oil Red O," _Medical Laboratory Technology_, Vol. 31:335-341, 1974.



Disclaimer: Energy Beam Sciences manufactures the JB-4 and JB-4A microtomes

for sevtioning plastic-embedded tissue, and sells GMA kits.



Best regards,

Steven E. Slap, Vice-President

********************************

Energy Beam Sciences, Inc.

Adding Brilliance To Your Vision

ebs@ebsciences.com

http://www.ebsciences.com/

The procedure that Steven posted does indeed work very nicly.....Provided that no alcohol was used

in the "dehydration" process. If you wish to look for lipids in GMA embedded tissues you must use

a series of graded (with water) monomer solutions instead of alcohols. Somewhere ????? I have the

procedure written but I can't put my hands on it right now (ie: I haven't got a clue as to where I

'filed' it). :(



regards,

Bob

Robert Schoonhoven

Laboratory of Molecular Carcinogenesis and Mutagenesis

Dept. of Environmental Sciences and Engineering

University of North Carolina

CB#7400

Chapel Hill, NC 27599

Phone

office 919-966-6343

Lab 919-966-6140

Fax 919-966-6123

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