Pewarnaan Aceto Orcein Dan Aceto Carmine

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 ac etic 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 a dd water into acid! Acetocarmine staining solution (1%) Carmine

Glacial acetic acid (45%) Ferric Chloride (FeCl2*6H2O) (10%) Optional Total volume

1

gram (g)

100 millilitre (ml) 5

millilitre (ml)

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 45 ml 5.0 gms STOCK 2 55 ml 45 ml 5 gms

Distilled water Acetic acid Carmine

Distilled water Acetic Acid 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. 2. 3. 4.

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