[Histonet] staining collagen

[John A. Kiernan]

Methyl blue, NOT methylene blue! Methyl blue is
one of the components of aniline blue, which is
often a mixture of related anionic
triphenylmethane dyes. (Methylene blue is a
cationic thiazine dye with very different staining
capabilities.) The picro-aniline blue methods pick
up thin collagen fibres that are missed by van
Gieson. The staining pattern is closely similar to
picro-sirius red F3B, but aniline blue does not
enhance the birefringence of type 1 collagen. 

John Kiernan
London, Canada.
----------------------------------------
Barbara Bublava wrote:
> 
> hi Marc,
> 
> pikro-aniline blue:
> 
> add 0,5g Anilinblue or methylenblue to 500ml  saturated aquous solution of
> picric acid.
> everything else is done like van Gieson
> Result: collagen, basement membranes and reticulin blue; cytoplasm yellow
> 
> John Kiernan´s book says reticulin and basement membranes are not stained if
> you use indigocarmine (0,5 g in 200ml) collagen should be blue-green in this
> case
> 
> greetings
> Barbara
> 
> ----- Original Message -----
> From: "Marc Tjwa" <marc.tjwa <@t> med.kuleuven.be>
> To: "Barbara Bublava" <barbara.bublava <@t> meduniwien.ac.at>
> Sent: Sunday, February 26, 2006 4:32 PM
> Subject: Re: [Histonet] staining collagen
> 
> Dear Barbara
> 
> I am also interested in the protocol that you
> have suggested. Would you please be so kind to
> send me the protocol by mail?
> 
> Tnx in advance
> 
> YS
> 
> Marc Tjwa
> 
> >You could use anilineblue instead of acid fuchsin. makes better contrast
> >and is described in John Kiernans book
> >
> >tell me if you can´t find a protocol
> >
> >greetings
> >Barbara, Vienna
> >----- Original Message -----
> >From: "Lobke De Bels" <Lobke.DeBels <@t> UGent.be>
> >To: <histonet <@t> lists.utsouthwestern.edu>
> >Sent: Friday, February 24, 2006 8:58 AM
> >Subject: [Histonet] staining collagen
> >
> >
> >
> >Von Gieson gives a good contrast for the human eye, but the computer can't
> >make
> >a difference between yellow/orange and red.
> >If I only use acid fushine (or aniline) everything stains pink(blue).
> >
> >I have to find a way to stain the connective tissue so the computer can
> >clearly
> >make a difference between connective tissue and the other tissue.
> >
> >The ideal should be a method that only stains collagen.
> >
> >
> >Many thanks in advance for any help
> >
> >
> >
> >Lobke De Bels
> >Department of Morphology
> >Faculty of Veterinary Medicine
> >Ghent University
> >Salisburylaan 133
> >9820 Merelbeke
> >Belgium
> >lobke.debels <@t> ugent.be
> >
> 
> --
> ============================================================
> Marc Tjwa, MD
> Center for Transgene Technology and Gene Therapy (CTG)
> Flanders Interuniversity Institute for Biotechnology (VIB)
> University of Leuven (KUL)
> Campus Gasthuisberg, Herestraat 49
> B-3000, Leuven, Belgium
> Tel: +32-16-345775; Fax: +32-16-345990
> E-mail: Marc.Tjwa <@t> med.kuleuven.be
> ============================================================