Albert Stain: Principle, Procedure & Results

Albert stain will help identify metachromatic granules found in Corynebacterium diphtheria in prepared slides from clinical specimens. This Corynebacterium is responsible for the disease diphtheria. Albert stain will stain and react only with metachromatic granules. The storage granules in this bacterium are called metachromatic granules because they exhibit the property of metachromasia, wherein the granules appear in a color other than the color used for staining.

Albert Stain

Albert Stain Principle

The principle of Albert stain is based on when the granules stain the purple-black against the light green counterstain cytoplasm. This helps to distinguish Corynebacterium diphtheria from most nonpathogenic dipatherdios which lack granules. Albert stain is made up of two staining solutions: Albert solution A and the second one is Albert solution B.

Albert Stain Composition

The composition of Albert stain is two types of mainly the first one is Albert stain A and the second one is Albert stain B. In Albert’s stain A, there are different types of components like Toluidine blue, Malachite green, Glacial acetic acid, and ethanol. The Alberts stain B has only two types of components Iodine and potassium iodide.

Albert Stain A

Toluidine blue and Malachite green, are basic dyes with a high affinity for acidic tissue like cytoplasm. When stained with Alberts stain A the volutin granules are stained blue and the cytoplasm is stained blue-green.

Toluidine blue  0.15 gm
Malachite green 0.20 gm
Glacial acetic acid1 ml
Alcohol (95% ethanol)2 ml
Composition of Albert Stain A Solution

Albert Stain B

Alberts stain B stains the blue-stained volutin granules blue-black.

Potassium iodide (KI)3gm
Composition of Albert Stain B Solution

Requirements For Albert Stain

  • Glass Slide: For the smear preparation.
  • Rack: For the help in slide standing.
  • Albert Stain A: Stain solutions.
  • Albert Stain B: Staining solution.
  • Blotting Paper: This is a highly absorbent type of paper used to absorb ink or oil.
  • Immersion Oil: This is a technique used to increase the resolving power of a microscope. 
  • Microscope: Help in finding.
  • Coplin Jar: Helps in holding microscope slides vertically during processing.
  • Spirit Lamp: Help in heating or staining.

Albert Stain Used For

Albert stain is used to detect the presence of metachromatic granules found in Corynebacterium diphtheriae. This will help to start proper treatment for you. Corynebacterium is responsible for the disease diphtheria.

Albert Stain Procedure

Albert stain steps:

  1. Prepare a smear on a clean slide.
  2. Air dry and heat fix the smear with the help of a spirit lamp.
  3. Place the slide on the rack with the smear upwards.
  4. Then put the Albert stain A into the smear.
  5. Allow the Albert stain 1 for 7 minutes.
  6. Drain the excess stain do not use water.
  7. Place Albert stain B for 1-2 minutes.
  8. Wash the slide with water and air dry.
  9. Add cedarwood/immersion oil to the smear.
  10. hen observe under the microscope at 1000x.

Albert Stain Result

  • The metachromatic granules stain bluish while the rest of the microbial cells stain green.
  • Corynebacterium diphtheriae is present in the sample it appears green-colored rod-shaped bacteria arranged at an angle to each other.

Albert Stain Precaution

  • Contamination may decrease the accuracy of the staining.
  • Safety precautions should be taken in handling, processing, and discarding all clinical specimens.
  • Samples should be processed in the correct containment level conditions.
  • Dispose of all material by local regulations.

Albert Stain Sample Collection

Need swab sample, take the sample as per the standard procedure. Bend the head backward and keep the mouth open till the sample is collected.


Reagents used in Albert’s staining procedure

Two types of reagents are used in the Albert stain procedure, Albert Stain A and Albert Stain B

How is Albert Stain different from other stains?

Albert Stain stands out due to its unique composition, offering distinct advantages in microscopic visualization compared to other staining techniques.

Is Albert Stain safe for human tissues?

Yes, when used correctly, Albert Stain is considered safe for human tissues, providing valuable information without causing harm.

Can beginners use Albert Stain in microscopy?

Absolutely! Albert Stain is user-friendly, making it accessible even for beginners in microscopy.

Are there alternatives to Albert Stain?

While alternatives exist, Albert Stain remains a preferred choice due to its effectiveness and versatility in various scientific applications.

What are the common mistakes in Albert Staining?

Common mistakes include improper timing and concentration. Following precise protocols is essential to avoid errors.

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