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.
To stain spirochete by using Becker’s method.
- Tartar sample.
- Fontana’s fixative.
- Fontana’s mordant.
- Becker’s stain.
- First of all a sterile pin is used to take a sample of tartar from teeth.
- The tartar sample is mixed and homogenized in sterile distill water and further used as a suspension.
- 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.
- The slide is further washed with water,air dried and treated with Fontana’s mordant for 5 minutes.
- After 5 minutes the mordant is drained off and slide is given a water wash treatment.
- Further the slide is treated with Becker’s stain for 3 to 5 minutes.
- Finally slide is washed with water,air dried and observe under oil immersion.
Flow Chart of Becker’s staining procedure
- 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.
- After chemical fixative the slide is treated with Fontana’s mordant this mordant contains tannic acid increases affinity of stain towards cell.
- Further slide is water wash to remove excessive mordant and treated with Becker’s stain for 3-5 minutes.
- This Becker’s stain is made up of basic fuchsin dissolved in alcohol.
- When this Becker’s stain is applied the alcohol evaporates the colloidal particles precipitates on cells and result in increases in size of spirochetes.
- Further the slide is water washed and dried ,observed under oil immersion lens.
After completion of staining procedure the spirochete are observed pink in colour.