Miller's stain for elastic fibres

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From:Katri Tuomala <> (by way of histonet)
To:histonet <>
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Hi everyone!
Since I had a few requests for the recipe, I'm posting it for anyone
interested. This is very similar, but not quite the same as the
Humberstone method Eric Kellar quoted.

Miller staining solution:

Victoria blue 4R            1g
New fuchsin                 1g
Crystal violet              1g

Dissolve above in 200 ml of hot distilled water (use a 600ml or bigger
beaker), then add the following:

Resorcin                    4g
Dextrin                     1g
30% Ferric chloride        50ml (freshly prepared)

Boil for 5 minutes, then filter while hot. Transfer precipitate plus the
filter paper to original beaker and re-dissolve in 200 ml of 95%
ethanol. Boil on a hot plate for 15 - 20 minutes (with filter paper and
all). Filter and make up to 200 ml with 95% ethanol. Add 2 ml
concentrated hydrochloric acid. This solution may be used repeatedly and
keeps several months.


1. Bring sections to water.
2. Oxidize in 0.5% potassium permanganate...5 min. Rinse in dist. water.
3. Decolorize in 1% oxalic acid ...2-3 min. Rinse in dist.water.
4. Rinse in 95% ethanol and transfer into staining solution .. 1-3
5. Rinse in 95% ethanol to remove excess stain.
6. Rinse well in dist.water and counterstain in van Gieson's ...5 min.
7. Dehydrate, clear and mount.

Results: Elastic fibres and mast cell granules .... black.
         Other tissue components as counterstained.

Reference: J.Med.Lab.Tech. vol 28, 1971 pg.148

Techs and pathologists are very happy with this method, since there is
no need for subjective differentiating step and results are consistent.

Katri Tuomala
St.Joseph's Hospital

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