ACID FAST STAIN

                                                            ACID FAST STAIN

Principle

This organism has a fatty capsule which influences the penetration and resistance to removal of stain by acid and alcohol. Phenolic acid and heat are used to reduce surface tension , increasing the porosity and forcing dyes to penetrate this capsule .


Solutions

 Strong carbol fuchsin
                                    Basic fuchsin              -           1 gm
                                    Absolute alcohol            -          10ml
                             5%Aqueous phenol     -           100ml.

Methylene Blue
                   Methylene Blue                -        1 gm
                   Distilled Water                 -        100ml

1% Acid Alcohol
                   Hydrchloric Acid              -        1 ml
                   70% Alcohol           -        99ml









Procedure

1)   Deparaffinise the section in xylene for 10 mts.
2)   Bring the section to water through descending series of alcohol.
3)   Flood section with carbol-fuchsin and heat to steaming, by intermittent flaming for 20 mnts.
4)   Wash it in Water.
5)   Differentiate with 1% acid alcohol until the section is light pink in colour.
6)   Keep it in running tap Water for 5 - 10 mnts.
7)   Counterstain with methylene blue for 30 see.
8)   Wash it in water. dehydrate, clean and mount.
ܝܚܚ
Result

Tubercle bacilli                 -        Pink

 Background          -       Blue

Comments

Post a Comment

Popular posts from this blog

PAS WITH AND WITHOUT DIASTASE

              PAS WITH AND WITHOUT DIASTASE Principle Section must be oxidized with periodic acid to convert 1:2 glycol group to          di-aldehyde in order that they will react with Schiff reagent to produce a colored end product. Glycogen is destroyed by the enzyme diastase which may be obtained from saliva or Malt Solutions Periodic Acid                 -           500mg Distilled Water               -           100 ml The solution is colourless All the glass wares should be properly cleaned and rinsed thoroughly in distilled Water. Procedure 1)    Add 1gm of basic fuchsin into 200 ml of hot distilled  water in a coni...
Read more

MELANIN BLEACH

                        MELANIN BLEACH Melanin is insoluble in almost all organic solvents. Strong oxidizing agent like potassium permanganate will bleach the melanin pigment. Solutions  1) 0.25% Potassium permanganate .                                     Potassium permanganate               -           250mg                                     Distilled water               ...
Read more

LUXOL FAST BLUE

                                      LUXOL FAST BLUE The  method is excellent for staining myelin fibres with good cellular differentiation. Solutions 1)     Luxol fast blue Solution . Luxol fast blue- 1 gm 95% alcohol-100 ml 10% acetic acid-5 ml 2)     Lithium Carbonate 0,05% Aqueous solution 3)     Haematoxylin .  Procedure 1)     Deparaffinise the section in xylene for 10 mts. 2)     Bring the section to water through descending series of alcohol. 3)     Stain in Luxol fast blue overnight at 37°C. 4)       Wash in 95% alcohol then in distilled water 5)     Commence differentiation by immersing sections in lithium carbonate solution for not more than 20 seconds 6)       Differen...
Read more