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SEMINAR ON PAS

PRESENTED BY :-Ms ANJANA B.Sc .MLT 3rd yrMODERATED BY:- Dr AMAR RANJAN

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PERIODIC ACID SCHIFFS REACTION :- [ PAS ]Detects intracellular glycogen and neutral

mucopolysaccharides

found in variable quantities in most hematopoieticcells.

Staining is seen in blast of acute lymphoblasticleukemia.

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Diffuse cytoplasmic positivity in

erythroleukemia.

Very useful in demonstrating the

abnormal glucocerebrosidaseaccumulation in GAUCHERS disease.

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PRINCIPLE PAS reaction depends upon the liberation of

carbohydrate radicals from the combination with

protein and their oxidation to aldehydes by the

schiff s reagent.

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RESULTS AND CONFIRMATION Positive reaction denotes the glycogen presence

Reaction can be confirmed by demonstrating that thepositive reaction disappears on treatment with diastase

or saliva before staining

other PAS positive material is changed by diastase

digestion.

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REACTION

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REAGENTS USED AND PREPARATION PERIODIC ACID SOLUTION 1g

DISTILLED WATER 100 ml

SHIFFS REAGENTCOUNTER STAIN

HEMATOXYLIN

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PREPARATION OF SCHIFFS Dissolve 1 g basic fuchsin in 200 ml of boiling, removingthe flask of water from the bunsen just before adding the

Basic fuchsin.

Allow the sol. to cool to 50c & add 2 g potassium

metabisulphite with mixing.

Allow to cool then add 2 ml conc. HCL, mix & add 2g

activated charcoal & leave overnight in the dark at room

temperature . Filter & store at 4c in amber coloured

bottle.

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FIX THE SMEAR INMETHANOL

TREAT WITH

PERIODIC ACIDFOR 10 MINUTES

RINSE INDISTILLED

WATER

COVER WITHSCHIFFS REAGENTFOR AN HOUR

WASH WELL INRUNNING WATER

COUNTER STAINWITHHAEMATOXYLIN

STAIN WASH AND RINSE

IN ALCOHOL

CLEAR INXYLENE MOUNT

IN DPX

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GIANT MULTINUCLEATE CELLSSHOWING GRANULAR POSITIVITY

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INTERPRETATION Normally all blood cells are (+ve) but

Erythroblasts (-ve)

Diffused stain pattern (Granulocytes)

- Granular stain (lymphocytes and monocytes)

- Plts deeply stained

- nRBCs (-ve) stain

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In diseases:

In CML

Lymphosarcoma

Hodgkins disease

nRBCsin M6

Thallasemia

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Positive PAS stain in ALL PAS positivity in M6. Not the intensestaining of the large abnormalerythroblast.

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CLINICAL SIGNIFICANCEPOSITIVITY IS SEEN IN :-

B CELLS :- Strongly +ve

T Cells :- Weakly +ve

Deep diffuse staining in erythroleukemia & in

thallasemia.

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Lesser degree of staining may be seen ina) Iron deficiency anaemia

b) Myelosclerosisc) Cord blood erythroblasts

d) Sideroblastic anaemiae) In various type of leukemia & hemolytic

anaemia.

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Reaction is +ve ina) Pernicious anaemia

b)Aplastic anaemia

c)

Lead poisoningd) Polycythemia Vera

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NOTE Reaction is best carried out on

a) Fresh blood

b) Bone marrow films

c) Old methanol fixed films

d) Films stained by Romanowsky dyes, months or before

can be quite satisfactory.

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THANK YOU