Re: [Histonet] aldehyde fuchsin

From:Geoff McAuliffe

  "Staining properties of aldehyde fuchsin analogues" by T.S. Buehner, 
G.S. Nettleton and J.B. Longley. 1979. J. Histochem. Cytochem. 
27(3):782-787. This paper discusses their results with paraldehyde, 
acetaldehyde, etc. Also, it is important that you prepare the aldehyde 
fuchsin from pararosanalin not just any old basic fuchsine (see Mowry et 
al., Stain Technology 55(2):91-103, 1980. And, the shelf life is short 
    One can also make aldehyde thionin .........


Marshall Terry Dr, Consultant Histopathologist wrote:

>Thanks John.
>Between you and Gayle, a most useful post concerning one of my tinctorially (if there is such a word) favourite stains.
>Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path
> Consultant Pathologist
> Rotherham General Hospital
> South Yorkshire
> England
>-----Original Message-----
>From: John A. Kiernan []
>Sent: 17 August 2005 17:36
>To: Marshall Terry Dr, Consultant Histopathologist
>Cc: Histonet
>Subject: Re: [Histonet] aldehyde fuchsin
>This is the method in M. Gabe's "Histological Techniques"
>(1976); it's also in my textbook. A similar method
>was published by Rosa,CC 1953 Stain Technol. 28:299-302.
>For comparisons of aldehyde fuchsine made in different 
>ways, see papers by GW Nettleton in J. Histochem.
>Cytochem. 1980-1982.
>Dissolve 1 g pararosaniline (CI 42500) in 200 ml water. 
>Heat to boiling and leave to cool to room temperature.
>Add 2.0 ml concentrated hydrochloric acid.
>Add 1 ml paraldehyde.
>Leave for 24 hours (longer if a pink ring
>appears when a drop of the solution is spotted 
>onto filter paper).
>Filter and discard the filtrate.
>Wash the residue with 50 ml water, then
>dry the filter paper and its contents in
>ab oven at 60C. Collect the dry aldehyde
>fuchsine powder in a little screw-cap
>The dye must be pararosaniline (CI 42500).
>The other components of basic fuchsine are
>not suitable for this stain. You can use
>acetaldehyde instead of its cyclic trimer, 
>paraldehyde. Acetaldehyde boils at 21C so
>it's less convenient to use, but unlike
>paraldehyde it isn't a controlled drug.
>Acetaldehyde is formed when paraldehyde
>reacts with acidified water.
>Staining solution.
>Aldehyde fuchsine powder 0.25 g
>70% ethanol              200 ml
>Glacial acetic acid      2.0 ml
>Leave overnight to dissolve. The
>solution does not need to be filtered.
>1. Stain hydrated paraffin sections, 5 minutes.
>2. Rinse in running tap water (messy).
>3. Immerse in 95% alcohol with 0.5% conc
>hydrochloric acid until no more colour
>is extracted (usually half a minute, but
>not critical; acid alcohol doesn't
>remove the real staining).
>4. Counterstain if desired (e.g. haemalum
>and fast green FCF).
>5. Wash, dehydrate, clear and coverslip.
>In some methods (eg for cystine-rich 
>proteins including insulin and classical
>neurosecretory material) an oxidizing step
>is put in before staining: 0.5% potassium
>permanganate in 2% sulphuric acid, 5 min,
>followed by a rinse in 1% oxalic acid to
>remove brown deposits of manganese dioxide
>from the sections, then wash in water.

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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