Aceto-Carmine Chromosome Stain Recipe

Non-iron version

1. Heat a solution of 45% acetic acid (45ml glacial acetic acid/55ml of distilled water) to
boiling
2. Add 0.5g of Carmine and continue heating for 15-20 minutes while stirring
3. Cool resulting solution
4. Filter to remove any precipitate

Iron containing version

1. Heat a solution of 45% acetic acid (45ml glacial acetic acid/55ml distilled water) to
boiling. ALWAYS use caution and wear personal protective equipment when in the
laboratory with concentrated acids (or any other time)
2. Add 0.5g of Carmine and continue heating for 15-20 minutes while stirring
3. Cool resulting solution
4. Filter to remove any precipitate
5. Separately make a solution of 45ml glacial acetic acid/55ml distilled water/5g Ferric
oxide)
6. Slowly (dropwise) add the Ferric oxide solution to 50ml of the Carmine solution until a
precipitate starts to appear
7. Promptly add 50ml of Carmine solution to the titrate mixture
8. Filter to remove any precipitate

Cautions: ALWAYS use caution and wear personal protective equipment when in the laboratory
with concentrated acids (or any other time)! Always add acid slowly into water and do not add
water into acid!

Acetocarmine staining solution (1%)

Carmine 1 gram (g)
Glacial acetic acid (45%) 100 millilitre (ml)
Ferric Chloride (FeCl2*6H2O) (10%)
5 millilitre (ml)
Optional
Total volume 100 millilitre (ml)
Note: Add carmine powder to boiling 45% glacial acetic
acid, cool rapidly, and then filter into a dark glass. Staining
can be intensified by adding ferric chloride (FeCl2.6H2O).
Caution: the stain stains clothes.
Aceto Carmine Stain ( Stern)

45 ml Glacial acetic acid

55 ml water
0.5 gm Carmine
Heat the 45% acetic acid solution to boiling. Add the 0.5gm
Carmine and continue heating for several minutes with
stirring. Filter and cool.

ACETO-CARMINE (BELLING'S)

STOCK 1
55 ml Distilled water
45 ml Acetic acid
5.0 gms Carmine
STOCK 2
55 ml Distilled water
45 ml Acetic Acid
5 gms Ferric oxide
(iron acetate)
Boil stock solution 1 for 15 minutes, cool and filter
To 50 mls of stock 1 add stock 2 until pcpt just appears
then quickly add 50 ml more of stock 1
***for most purposes omit stock solution 2 and just use
stock solution 1.

Acetic Orcein Stain Recipe

1. Add 1g of Orcein to 45ml of glacial acetic acid
2. Heat solution from step 1 to dissolve
3. Slowly add solution from step 2 into 55ml of distilled water
4. Allow to cool and store

Cautions: ALWAYS use caution and wear personal protective equipment when in the laboratory
with concentrated acids (or any other time)! Always add acid slowly into water and do not add
water into acid!

Aceto-orcein staining solution (1%)

Orcein 1 gram (g)

Glacial acetic acid 55 millilitre (ml)
ddH2O to 100 millilitre (ml)
Total volume 100 millilitre (ml)
Note: Pour 55 ml boiling glacial acetic acid over 1 g orcein
powder. Cool the solution, add 45 ml of distilled water and
filter. Prepare fresh before use.