[Histonet] Acriflavine staining of alginate for mucopolysaccharides

[Sujata Sovani]

Hi All,

I am new to histology.
I came across the protocol below for staining GAGs (I plan to use that for alginate) in an online book 'Theory and practice of histological techniques', By John D. Bancroft, Marilyn Gamble. The protocol is by Hollander dated 1963.

Would like to know if anyone has used this protocol before? Any tips/specific details might be helpful.
The 20% HCl - does it mean 20% of 37.3% concentrated hydrochloric acid?
Any details about acriflavine to be used would be helpful.

Thank you.
Regards,
Sujata

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The protocol is as below:

Protocol

Acriflavine-DMAB method for sulfatide (Hollander 1963)

Fixation and sections
Post-fixed cryostat sections; formal calcium-fixed frozen sections

Prepa­ration of reagents

1.Acriflavine stock solution
Acriflavine                                          100mg
Distilled water at 80 deg C                  20ml

Store in dark at 4 deg C

2.Acriflavine working solution
  0.1M citrate-HCl buffer pH 2.5       99ml
  Stock acriflavine solution                       1ml

3.DMAB solution
   p-dimethylaminobenzaldehyde            0.6g
   20% HCl                                          30ml
   Isopropanol                                      70ml

Method


 1.  Mount sections onto slides
 2.  Stain for 6 minutes in acriflavine solution
 3.  Differentiate for 1 minute in two changes of 70% isopropanol
 4.  Treat with DMAB reagent for 30-45 seconds
 5.  Rinse in distilled water for 2-3 minutes
 6.  Counterstain nuclei in Mayer’s or Carazzi’s hematoxylin
 7.  Blue in tap water, rinse in distilled water and mount sections in glycerin jelly.