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FIXATION:        10% buffered neutral formalin.

TECHNIQUE:        Cut paraffin sections at 6 μm.

SOLUTIONS:

Crystal Violet Solution

Crystal violet, C.I. 42555--------------------------------------------------------                1.4  gm

Alcohol, 95% ------------------------------------------------------------------                10.0 ml

Distilled water -------------------------------------------------------------------        300.0 ml

Hydrochloric acid, concentrated -------------------------------------------------        1.0 ml

*Apathy’s Mounting Media

Acacia (gum arabic) ------------------------------------------------------------                50.0 gm

Sucrose (can sugar) -------------------------------------------------------------                50.0 gm

Distilled water -------------------------------------------------------------------        150.0 ml

Sodium chloride -----------------------------------------------------------------        10.0 gm

Thymol -------------------------------------------------------------------------                0.1 gm

       Mix the acacia, sucrose and distilled water in a 500 ml beaker using

       a hot plate stirrer.  Heat the solution to 58o-60o C.  It usually takes

       5-6 hours for the acacia to completely dissolve.  Add distilled water

       as needed to maintain the original volume of the solution.  Dissolve

       the sodium chloride and thymol in the solution.  Filter through Pyrex

       glass wool fiber.  Store in a refrigerator at 3o-6o C.  Sodium chloride

       prevents “bleeding” and thymol acts as a preservative.

• Use control slide.

1. Deparaffinize and hydrate to distilled water.
2. Stain in working crystal violet solution for five minutes.
3. Wash in running tap water for five minutes and rinse in distilled water.
4. Mount with Apathy’s mounting media.

Amyloid ----------------------------------------------------------                purplish violet

Other tissue elements ---------------------------------------------                blue

*”Bleeding” or diffusion into the surrounding mounting media of basic aniline dyes, tends to occur with aqueous mounting media.  Because this Apathy’s mounting media has been modified form the original formula by the addition of sucrose and sodium chloride, diffusion of the dye does not occur.

This crystal violet method does not stain all types of amyloid as well as Puchtler’s Congo red technique.  However, it usually better demonstrates amyloid in heart muscle than the Congo red method.

Lieb, E.:   Permanent Stain for Amyloid, Amer. J. Clin. Path. 17:413-414, 1947.