Wednesday 16 September 2015

Staining of Metachromatic granules by Albert's staining method.

Hello to all lovelies..
Hope you are doing well.
I know I have been neglecting our staining techniques since a long time and we all know how basic and important they are. And that's why I am here.
Which staining I was gonna discuss today?
...
...
Oh yeah, Albert's :D
I don't know why most of the staining techniques were invented by males, I mean it's the females who are more considered about the stains and staining since time started. You know what I mean, right?
Hey girl, here Take this straw....Now suck it up and start with the content.
Sure let's go.



What is this staining technique all about?
This technique is about staining metachromatic granules also known as volutin granules. It requires two different stains named Albert's stain 1 and Albert's stain 2. It is a differential staining techniques just like gram staining or capsule staining. It imparts different colors to the bacterial cells and the granules.
What are metachromatic granules?
Metachromatic granules are present in Corynebacterium diphtheriae. They are present inside the cytoplasm of the cell and function as storage bags for organic polyphosphate. They are called metachromatic granules because they display different colors. The cells take up the green color whereas the granules are seen in red.
How do I prepare Albert's stains?
Here are the reagents that you will require-
Albert's stain 1:
Toluidine blue                  0.15gm
Malachite green.              0.20 gm
Glacial acetic acid           1 ml
Alcohol(Ethanol)            2 ml

Dissolve the dyes in alcohol and then mix it with water and glacial acetic acid.
Filter the stain using after 24 hrs to eliminate any precipitates.
Make up the volume to 100 ml with distilled water.

Albert's stain 2-
Iodine                              2 gm
Potassium iodide.            3 gm

Now some sulking examiners who have nothing to do in life might expect you to remember this.
So easy peasy lemon squeezy, here's a mnemonic for it.


How to perform this staining?

  1. Prepare a smear on a clean, grease free slide. Having a glass is enough according to me, who cares for the grease...nah, just kidding..or was I?
  2. Dry and heat fix.
  3. Cover the stain with Albert's 1 and keep it for 5 mins.
  4. Drain the stain and cover the smear with Albert's iodine for 2 mins.
  5. Drain and wash under tap water. Note: keep the tap low:p
  6. Air dry and observe under oil immersion.
How do they look?
Well to me, they look like dumbells, because the straight cells are flanked on the two sides with round shaped granules. They cells though sometimes are straight or V shaped or L shaped.

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