Re: [Histonet] MacNeal's tetrachrome

From:kbowden



5% MC NEAL'S STAIN
from the lab of Dr. Robert Schenk
Bern Switzerland
Milled/Surface Stain

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.
Distilled Water                                                        
    42.5 ml.
   Stir on Stir plate, filter afterwards


0.1% Basic Fuchsin

Basic Fuchsin                                                            
0.5 gm
Deionized water                                                        
500.0 ml


0.1% Formic Acid

Formic Acid                                                            
   1.1 ml
Deionized water                                                        
1000.0 ml

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
Methyl Alcohol                                                        
    500 ml
Glycerin                                                                
        500 ml
   Heat at 50C for 1 day, then 37C for 3 days in oven (capped).  
Filter and store in dark bottle in dark.


Results:
    bone    - reddish/purple
    cells/nuclei    - blue
    Cartilage    - blue/purple
    ligamenti, connective tissue    - pinkish/red

--
Karen Bowden
Staff Research Associate II
University of CA, San Diego
Department of Orthopedics
9500 Gilman Dr. 0630
La Jolla, CA 92093-0630
858-534-4655 voice
858-534-5304 fax


CONFIDENTIALITY NOTICE:
THE INFORMATION TRANSMITTED IN THIS E-MAIL IS INTENDED ONLY FOR THE
PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL
AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR
OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON THIS INFORMATION
BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED.
IF YOU RECEIVED THIS E-MAIL IN ERROR, PLEASE CONTACT THE SENDER AND
DELETE THE MATERIAL FROM ANY COMPUTER.


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,
> Orla
> ******************************
>
> 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
> www.sheffield.ac.uk
>
> E-mail:o.m.gallagher@sheffield.ac.uk
> Tel:   0114 271 3783(lab)
>        0114 271 3337(office)
> Fax:   0114 271 1711 
>
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>   
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet




<< Previous Message | Next Message >>