[Histonet] pararosaniline for aldehyde fuchsin

[Gayle Callis]

Sigma has an excellent pararosaniline for this purpose

At 08:27 AM 8/18/2005, you wrote:
>  "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 .........
>
>Geoff
>
>
>>
>>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
>>bottle.
>>
>>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.
>>
>>Method.
>>
>>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 mcauliff <@t> umdnj.edu
>**********************************************
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>
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)