Spirochete Staining by Becker’s Method

Spirochete Staining by Becker’s Method

  • Spirochete are spiral shape bacteria .Becker’s staining is another special staining used to stain spirochete.
  • Other Spirochete staining techniques are Rue’s method and Levaditi’s method.

Aim

To stain spirochete by using Becker’s method.

Requirement

  1. Tartar sample.
  2. Fontana’s fixative.
  3. Fontana’s mordant.
  4. Becker’s stain.

Procedure

  1. First of all a sterile pin is used to take a sample of tartar from teeth.
  2. The tartar sample is mixed and homogenized in sterile distill water and further used as a suspension.
  3. A drop of suspension is taken on slide ,smear is prepared,air dried and  fixed by using chemical fixative that is Fontana’s fixative for 30 seconds for 2 times after interval of 1 minute.
  4. The slide is further washed with water,air dried and treated with Fontana’s mordant for 5 minutes.
  5. After 5 minutes the mordant is drained off and slide is given a water wash treatment.
  6. Further the slide is treated with Becker’s stain for 3 to 5 minutes.
  7. Finally slide is washed with water,air dried and observe under oil immersion.

Flow Chart of Becker’s staining procedure

Flow chart of Becker's spirochete staining procedure
Flow chart of Becker’s spirochete staining procedure

Mechanism

  1. The smear is prepared and air dried,due do delicacy of cells the smear is fixed by using chemical fixative that is Fontana’s fixative.
  2. After chemical fixative the slide is treated with Fontana’s mordant this mordant contains tannic acid increases affinity of stain towards cell.
  3. Further slide is water wash to remove excessive mordant and treated with Becker’s stain for 3-5 minutes.
  4. This Becker’s stain is made up of basic fuchsin dissolved in alcohol.
  5. When this Becker’s stain is applied the alcohol evaporates the colloidal particles precipitates on cells and result in increases in size of spirochetes.
  6. Further the slide is water washed and dried ,observed under oil immersion lens.

Result

After completion of staining procedure the spirochete are observed pink in colour.

 

 

 

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