Flagella Staining by Loeffler’s Method and its Details

Flagella Staining by Loeffler’s Method and its Details

Flagella is the most important locomotory organ of a bacterial cell and for studying this structure it is important to stain the flagella.So here we have another staining technique used for staining of flagella that is Loffler’s staining technique.

There are some other staining techniques that are also used for staining of flagella that are Bailey’s method,Gray’s method, Casares-Gil’s method and Leifson’s method.

Here in this article we are going to study  Loffler’s method in detail.

Aim

To stain flagella of a cell by using Loffler’s method.

Requirement

  1. Flagellated cell culture.
  2. Loeffler’s mordant.
  3. Loeffler’s stain.
  4. Distill water.
  5. Chromic acid.

Procedure

  1. The slides are cleaned by boiling them in chromic acid solution.
  2. The smear is prepared as given in the flow chart below.
  3. Further after smear preparation the slide is treated with Loeffler’s mordant and heated it till steam appears approximately it takes 3 minutes.
  4. After that slide is gently washed by distill water.
  5. Further slide is treated with Loeffler’s stain and heated till steam appears.
  6. Finally slide is washed with water,air dried and observe under oil immersion.

Flow chart for smear preparation of flagella staining.

Flow chart of smear preparation for Flagella staining

Flow chart of flagella staining procedure by Loeffler’s method.

Flow chart of flagella staining procedure by Loeffler's method

Mechanism

  1. As we know flagella of a bacterial cell are very thin,hair like structure .This delicate structure can easily get destroyed so here there is no heat fixation step.
  2. Now it is very important to increase the thickness of flagella so that we can stain it and observe it under microscope.
  3. So here in this staining technique we use Loeffler’s mordant this mordant contains some colloidal particles with alcohol base.
  4. Now when we apply this mordant and heat it the alcohol get evaporated and the colloidal particles get adhere to the flagella and increase its thickness.
  5. Further this smear is treated with Loeffler’s stain.This stains precipitate on flagella and thus stains the flagella as well as cell in pink colour.

Result

Here Flagella as well as bacterial cell get stain in pink colour.

 

 

 

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