Re: Rhodanile blue (Used by? & Discussion)
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From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> (by way of histonet) |
To: | histonet@histosearch.com |
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On Thu, 9 Dec 1999, Lesley Weston wrote:
> Does anyone happen to remember the recipe for rhodanile blue? I've lost
> mine and can't find it on the net either. Thanks.
My short answer is, "No, I've not got the recipe," but Lesley's
enquiry raises some interesting points. How much is this stain
used, for what purposes, and with what success?
This was a concoction of Edward Gurr ("Encyclopaedia of Microscopic
Stains" and various other books). The following notes are from
long-term memory and Lillie's 10th edition of Conn's Biological
Stains (1977), which is to hand. Lillie cites MacConnail & Gurr,
Irish J. Med. Sci. 1964: 243-250 for the seminal work on rhodanile
blue. I haven't seen this paper, but have made a note to look it
up.
It seems likely that Gurr mixed a cationic dye (nile blue) with an
amphoteric (but predominantly cationic) dye, rhodamine B. He published
a formula, presumably speculative, that implied the formation of
an amide from the -COOH of rhodamine B and the -NH2 of nile blue, to
make a cationic xanthene-oxazine hybrid dye. This is chemically
improbable if the parent dyes were combined in aqueous solution, and
it is very much more likely that solutions contain coloured cations
of rhodamine B (red), nile blue (blue) and perhaps also some nile
red, which is a hydrophobic product of oxidation of nile blue.
Lillie (1977) reviewed 4 or 5 published applications of rhodanile
blue, but gave no indication of how the results were influenced by
such things as the pH solvents during and after staining.
Edward Gurr published other dye mixtures as "compounds," notably
FALG (and FALGOG). These contained acid fuchsine ("fuchsinic acid"
in Gurr's terminology) with light green (and orange G). They were
touted as one-step multicolour reagents. I tried these soon after
Gurr's "Encyclopaedia" came out, but could get only grey-violet
shades with the occasional bright red cell or unrecognizable blob.
Please don't interpret the preceding paragraphs as an attack on the
memory of the late Edward Gurr, who sold good stains and wrote good
books. ("When I am dead let it be said, That his sins were scarlet
but his books were read." - Hillaire Belloc) Perhaps some
HistoNetter older than I will tell us more about the saga of the
Brothers Gurr.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
E-mail: kiernan@uwo.ca
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