Re: Casteneda's stain

From:Bryan Hewlett


Your original posting requested a reference for this staining method.
I checked but could not find one!

However, you now seem to be requesting the method, that I do have.
It may be found in "Biological staining methods"  6th edition by GT Gurr or
in ' Medical Laboratory Technology and Clinical Pathology" 2nd edition 1969,
p982. Lynch, Raphael, Mellor, Spare and Inwood.

Casteneda's method for Rickettsiae in smears.

Solutions required.
A) buffer(pH7.5) Potassium dihydrogen phosphate 1.0g, disodium hydrogen
phosphate 12 H2O 25.0g, water 1L add 1.0mL formalin.
B) 1% w/v Methylene blue in methyl alcohol.
C) 0.2%w/v aqueous safranin 'O' 25mL, 0.1% aqueous acetic acid 75mL

1 Air dry thin smears.
2 stain for 3 minutes in freshly prepared mixture of 20.0mL buffer; 1.0mL
formalin and 0.15mL solution B.
3 drain(don't wash) slide
4 Treat with counterstain C for 2-5 secs
5 wash in tap water and dry.

Rickettsia...... blue
Cells..... red

If you wish to stain Rickettsia in tissue sections, I would recommend Giemsa
or Macchiavello's  or Mann's methyl blue/eosin.

Hope this helps


----- Original Message -----
From: "sue hacker (IAH-C)" 
Sent: Friday, March 21, 2003 4:05 AM
Subject: Casteneda's stain

> Dear All,
> Just trying this again, Is there anyone out there in Histonet land who has
come across Casteneda's stain and has any reference no matter how small to
this method.
> I have found a paper for a method used by Casteneda for Brucellosis dated
1947 but that is not the correct one.
> I am looking for Casteneda's stain for Rickettsiae.
> Desperation setting in!!
> Thanks,
> Sue Hacker
> IAH Compton,
> Berkshire.UK.

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