5% MC NEAL'S STAIN
from the lab of Dr. Robert Schenk
1. Agitate in 50% EtOH 10 to 15 min.
2. Distilled water wash 3 min.
3. Agitate in 0.1% Formic acid 5 to 10 min.
4. Distilled water wash 3 min.
5. Distilled water wash 3 min.
6. Distilled water wash 3 min.
7. 5% Mc Neal's (Filtered) 5 min.
8. Distilled water wash 3 min.
9. Distilled water wash 3 min.
10. Distilled water wash 3 min.
11. 70% EtOH 1 dip
12. 0.1% Aqueous Basic Fuchsin (Filtered) 15 to 30 sec
13. Distilled water wash 3 min.
14. Distilled water wash 3 min.
15. Distilled water wash 3 min.
16. Air dry
5% Mc Neal's Stain
Mod. Mc Neal's Tetra chrome (KEEP IN DARK) 5 ml.
0.1% Toluidine Blue aqueous 2.5 ml.
Stir on Stir plate, filter afterwards
0.1% Basic Fuchsin
0.1% Formic Acid
10% Mod. Mc Neal's Tetrachrome (STOCK)
Methylene Blue Chloride (Merck 6045-color 52015) 1.0 grams
Azure A Eosinate (Chroma 1A 328) 1.6 grams
Methyl Violet, Bernthsim (Chrome 1A 098) 0.2 gram
Heat at 50°C for 1 day, then 37°C for 3 days in oven (capped).
Filter and store in dark bottle in dark.
bone - reddish/purple
cells/nuclei - blue
Cartilage - blue/purple
ligamenti, connective tissue - pinkish/red
Staff Research Associate II
University of CA, San Diego
Department of Orthopedics
9500 Gilman Dr. 0630
La Jolla, CA 92093-0630
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Orla Gallagher wrote:
> Dear Histonetters,
> Our lab would like to try staining some undecalcified LR White-
> embedded bone sections with MacNeal's tetrachrome stain. I've
> found a recipe on the Histonet archive which mentions making up
> the stain from scratch but doesn't say how long to stain for or how
> to treat the sections after staining. The recipe also mentions
> etching in formic acid before staining. Has anyone got some advice
> as I haven't tried any etching techniques before?
> Many thanks,
> Ms. Orla Gallagher
> Academic Unit of Bone Biology
> D Floor (DU20) Medical School
> Henry Wellcome Laboratories for Medical Research
> University of Sheffield
> Beech Hill Road
> Sheffield S10 2RX
> Tel: 0114 271 3783(lab)
> 0114 271 3337(office)
> Fax: 0114 271 1711
> Histonet mailing list
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