Flagella Staining Technique by Liefson’s Method

Flagella Staining Technique by Liefson’s Method


  • Bacteria have two types of locomotory organs and that are Flagella and pili.
  • Here we are studying about flagella staining.
  • Flagella is a thin,hair like structure made up protein called as flagellin.
  • It size ranges from 20 µ to 200 µ in length.
  • Flagella is one of the most important locomotory organ .It is mainly made up of three parts- 1) Basal body 2) Filament 3) Hook.
  • Flagella is generally present in rod shape bacteria and very few cocci shape bacteria posses flagella.
  • As flagella are very thin and hair like they cannot be easily observed under microscope.
  • So a special technique is design to increase thickness of flagella as well as stain it.
  • Due to these technique we can observe structure of flagella easily under microscope.

Please check out the below video for Flagella staining by Liefson’s method.


To Stain Flagella of cell by using Leifson’s method


  1. Flagellated cell culture slant.
  2. Leifson’s stain.
  3. 1 % Methylene blue.
  4. Distilled water.


  • First of all take two hours old flagellated cell culture slant and add two to three drops of sterile distill water in the slant with the help of sterile pipette.
  • Note that the distill water is added slowly without disturbing the growth of cells.
  • After addition of distill water incubated the slant for 20 minutes.
  • Then take a drop of suspension from the slant and place the drop on a clean slide which is kept in slanting position.
  • The drop should flow slowly from one end of slide to other end to avoid folding of flagella on cell.
  • Allow smear to air dry here we don’t use heat fixation treatment .
  • After air drying the slide is flooded with Leifson’s stain till a thin film of shinny surface appear.
  • After this give a gentle stream of water wash treatment to a slide.
  • Now treat the slide with 1 % methylene blue treatment for 1 minute.
  • Give the slide water wash treatment ,air dry and observe under oil immersion lens.

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