[Histonet] Alcianblue for fast frozen sections

Tony Henwood (SCHN) tony.henwood at health.nsw.gov.au
Sun Jan 29 08:00:06 CST 2023

Hello Gudrun,
Alcian blue for fast frozen sections

I was not aware that alcian blue would be routinely used on frozen sections of transplant liver biopsies.
There are other staining techniques that contain a blue dye, that have been used in liver biopsies including Masson's Trichrome (aniline blue) and Millers Stain (Victoria blue). Sulphated Alcian Blue has been used for the demonstration of amyloid in liver biopsies, though not usually on transplant liver biopsies.
Alcian blue has been used on frozen sections to demonstrate carboxylated mucins (J Clin Pathol 42(1): 101-105, 1989).

Alcian blue staining has been used on FFPE sections of transplant liver sections to show loss of heparan sulphate from endothelial cells of liver graft vessels. Alcian blue at pH 5.8 with 0.3 M MgCl2 on 3-micrometre paraffin-embedded liver section. At this MgCl2 concentration only sulfate glycosaminoglycans such as heparan sulfate stain. Loss of heparin sulphate reduces the ability of the graft to maintain an anti-coagulant environment and can result in diffuse microvascular thrombosis and vascular congestion (see Remuzzi et al (2005). Hemolytic uremic syndrome: a fatal outcome after kidney and liver transplantation performed to correct factor H gene mutation. American Journal of Transplantation, 5(5), 1146-1150).
If this is the alcian blue technique needed, then the question arises whether heparan sulphate is insoluble, and therefore stainable, following cryotomy and fixation. Heparan sulphate has a solubility of 100mg/mL in water but is insoluble in ethanol. I would suggest validating the technique on frozen sections of kidney and liver, comparing the staining results to corresponding FFPE sections.

Please tell us how you go.


Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA)
Principal Scientist, the Children’s Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney
Tel: 612 9845 3306
Fax: 612 9845 3318
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA
From: Gudrun Lang via Histonet <histonet at lists.utsouthwestern.edu>
Sent: 28 January 2023 04:38
To: histonet at lists.utsouthwestern.edu <histonet at lists.utsouthwestern.edu>
Subject: [Histonet] Alcianblue for fast frozen sections

Dear histonetters!

Today I've heard about alcianblue staining on fast frozen sections, but I've
got no details.

I would like to know, if the staining result is the same as for staining AB
on paraffinslides.

They use the stain on transplantation liver.

Is this a usual procedure?

I would be glad about any information.

Thanks in advance


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