Perls Prussian Blue

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Perls Prussian Blue

• Sree Puliyampatta

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Perls A German Pathologist Perls Max
(1843-1881) who first suggested this
methodPrussian A dark blue pigment (ferric
blue hexacyanoferrate)


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Perls prussian blue test
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The very two commonly tested tissues 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 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

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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)

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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

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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

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Result

• Ferric saltsDeep 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 the routine
• Toxic fumes
• Safety Should be very careful in setting time
  and power
• Proper laboratory microwave is expensive

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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