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Perls Prussian Blue

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Perls Prussian Blue

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    1. Perls Prussian Blue Sree Puliyampatta

    2. Perls: A German Pathologist Perls Max (1843-1881) who first suggested this method Prussian A dark blue pigment (ferric blue: hexacyanoferrate)

    3. Perls prussian blue test

    4. The very two commonly tested tissues for Perls in our lab Liver core non neo Bone marrow (trephine)

    5. Perls Test for what? To detect the presence of ferric iron in tissues Normal iron content: 3-4 grams in the body If the content exceeds beyond 15 grams, the condition is known as iron overload.

    6. Iron overload..some facts A condition such as accumulation of iron in the body: due to.. Hereditary disease (hemochromatosis), mutated genes: Daily iron absorption is greater than the amount needed : Normal body cannot increase iron excretion. Excess iron will be deposited in joints, liver, heart, pancreas etc., causing organ damage. Mutated genes disrupts iron homeostasis. Iron overload through blood transfusion: Chronic anemics due to inborn errors (thalassemia, sickle cella anemia) . Transfusion therapy increases the life span. Iron overload is an inevitable consequence I.e., iron from the senescent transfused RBC released in to circulation and then to body organs. Visual signs: Bronze or slate gray skin pigmentation

    7. Perls test technical Principle: Ferric iron is stored with in a protein called ferritin. Dilute mineral acid releases ferric ions from protein. Ferric ion in the presence of ferrocyanide ions will precipitate in to a complex, potassium ferric fero cyanide giving the blue colour (Prussian blue)

    8. Method (routine) Dewax to water, both test and known positive control Mix equal volumes of 2 per cent hydrochloric acid and 2 per cent of potassium ferrocyanide solution and flood over the sections on a rack for 10 mins. Wash well in several changes of water Counter stain with NFR Wash dehydrate and mount

    9. Method (microwave) Dewax to water, both test and known positive control Place slide in plastic staining jar ( slide mailing box would be fine) containing 16 ml Perls working solution (8ml of HCl and 8 ml of pot.ferrocyanide) and microwave at 450 W for 15 secs. Wash well in several changes of water Counter stain with NFR Wash dehydrate and mount

    10. Result Ferric salts……………Deep blue Nuclei…………………..Red

    12. Microwave advantage/disadvantage Quick 15 seconds over 10 minutes Microwave electrical hazards Usage of more chemicals than the routine Toxic fumes Safety: Should be very careful in setting time and power Proper laboratory microwave is expensive

    13. Questions….. Total Perls test done in MMH the year 2007 Jan to Dec 206 For convenience it is advisable to mix the solution A and B and keep it..If no why? Find the answer or answer on next talk

    14. Thank you

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