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Perls Prussian BlueSree Puliyampatta

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 Perls: A German Pathologist

Perls Max (1843-1881) who

first suggested this method

Prussian A dark blue pigment (ferric 

blue: hexacyanoferrate)

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Perls prussian blue test

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The very two commonly testedtissues for

Perls in our lab

Liver core non neo

Bone marrow (trephine)

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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.

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Iron overload..some facts

A condition such as accumulation of iron in thebody: 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 consequenceI.e., iron from the senescent transfused RBC released in to circulation

and then to body organs.

Visual signs: Bronze or slate gray skin pigmentation

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Perls test technicalPrinciple: 

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 ferocyanide giving the blue colour (Prussian blue)

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Method (routine) Dewax to water, both test and known positive

control

Mix equal volumes of 2 per cent hydrochloric acidand 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

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Method (microwave) Dewax to water, both test and known positive

control

Place slide in plastic staining jar ( slide mailing boxwould 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

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Result

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

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Ferric salts

Nucleus

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Microwave advantage/disadvantage

Quick 15 seconds over 10 minutes

Microwave electrical hazards

Usage of more chemicals than theroutine

Toxic fumes

Safety: Should be very careful insetting time and power

Proper laboratory microwave isexpensive

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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

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Thank you