[Histonet] Differentiation between Eosinophil and Neutrophil by SiriusRed staining?

[John Kiernan]

Bryan Llewellyn's method uses sirius red F3B from
an alkaline solution, in contrast to the methods for
collagen (Puchtler, Junqueira etc) in which the same
dye is dissolved in saturated aqueous picric acid.

An alkaline solution of an anionic dye will be
attracted only to strongly basic proteins - those
rich in arginine. (The guanidino side-chain of
arginine is a cation even at high pH.) This limits
the staining to eosinophil granules, Paneth cell
granules and the tails of spermatozoa. Amyloid is 
also stained - the principal application of the
method - but that's for different reasons. Alkaline
solutions of some anionic dyes can stain elastic
fibres and laminae (also for different reasons).
Stained eosinophils are unlikely to be confuesed 
with any of the other things that can be coloured 
by an alkaline solution of an acid dye. The dye
doesn't have to be sirius red F3B. Eosin Y will
do the job; that's how the cells got their name.

John Kiernan
Anatomy, UWO
London, Canada.
______________________________________________
Felix Rintelen wrote:
> 
> Hi everyone,
> 
> 
> 
> I'm trying to decide by histological staining whether an infiltrate in mouse tissue is dominated by either neutrophils or eosinophils. I performed the Llewllyn's sirius red F3B staining (see web page http://stainsfile.info/StainsFile/stain/amyloid/siriusllew.htm, Sirius Red F3B = Direct Red 80 was from Sigma Cat N° 365548) which according to the protocol stains Eosinophils and Paneth cells. This staining was also recommended in the Histonet and indeed the staining worked very nicely.
> 
> Now, my question is just, if anybody knows if neutrophils are not stained by this procedure or not. In other words, if I can be sure that when a cell is stained red, it will for sure be an eosinophil and not a neutrophil.
> 
> 
> 
> Thank you very much in advance!
> 
> 
> 
> Best regards,
> 
> 
> 
> Felix
> 
> 
> 
> 
> 
> Felix Rintelen
> 
> Serono Pharmaceutical Research Institute
> 
> Geneva, Switzerland
> 
> 
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