Albert staining of metachromatic granules ideal for Corynebacterium diphtheriae

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Albert stain MLTGEEKS

This staining technique is used to stain the volutin or metachromatic granules of  corynebacterium diphtheriae,other corynebacterium species and occasionally Bacillus species especially when the organism is cultured on a protein rich medium such as dorset eggs,loeffler serum or Egg yolk agar which makes the volutin granule most numerous.

Principles of Albert staining technique

This stain demonstrates the presence of metachromatic granules which are found in Corynebacterium diphtheriae and other corynebacterium species.

  • The granules stain purple-black against the light green counterstained cytoplasm.
  • This helps to distinguish Corynebacterium diphtheriae from most of the short nonpathogenic diphtheroids species lacking volutin granules

Albert’s stain A (Composition)

IngredientsGrams  and ml
Toluidine blue0.150 gm
Malachite green0.200 gm
Glacial acetic acid1.000 ml
Ethyl alcohol, 95%2.000 ml
Distilled water100.000 ml

Albert’s stain B (Composition)

Iodine crystals2.000 gm
(S002) Iodine crystals3.000 gm
Distilled water300.000 ml

Material Required for Albert staining technique

  • Albert’s stain A
  • Albert’s stain B
  • Inoculum or specimen
  • Clean slide
  • 70% alcohol

Procedure for Albert staining technique

  • Make a thin smear of the specimen on a clean slide
  • Allow to air dry
  • Fix the dried smear with 70% alcohol or with gentle heat
  • Cover the dried smear Albert’s Stain A for 3-5 minutes.
  • Do not wash instead drain the slide
  • Flood the smear with Albert’s Stain B for 1 minute
  • Rinse the slide with  clean water
  • Blot dry the slide with tissue paper and examine  starting with 40X objectives in order to check the staining and the distribution of the cells then switch to 100X  oil immersion objective
Also read  Egg Yolk Agar Base Selective and Differential Medium for the Isolation of Clostridium perfringens

Observation and interpretation

ObservationStain’s appearance
Bacteria cellsPale green
Volutin or metachromatic granulesBlack

The metachromatic granules of Diphtheria bacilli  will stain black while their cytoplasm stains light green or green


  • Monica cheesbrough Part 2,p44.

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About the Author: Arthur Westmann

DEFFE ARTHUR (AMOEBAMANN) is the founder and author of MLTGEEKS and MLTEXPO.He’s from Cameroon and is currently a Final year State Medical Laboratory Technician (MLT MA). Beyond lab works, he’s a passionate internet user with a keen interest in web design and blogging. Furthermore He likes traveling, hanging around with friends and social networking to do in his spare time.

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