Simple staining method in microbiology
Staining method, technique, or procedure that is commonly employed in microbiology labs. Only one dye is utilized throughout the staining procedure in this form of staining. It's most commonly used to show bacterial shape and organization.
The dye stains the bacteria or its components.
Carbol fuchsin stain, for example.
Blue methylene stain Stain of crystal violet.
Procedure
1. Make a smear and label it.
2. Allow the smear to dry in air.
3. Fix the smear over a flame.
4.Apply a few drops of positive simple stain like 1% methylene blue, 1% carbolfuchsin or
1% gentian violet for 1 minute.
5. Wash off the stain with water.
6. Air-dry and examine under the oil immersion objective.
Simple and differential staining methods in microbiology
1. Basic staining
Is a staining method, technique, or procedure that is commonly employed in microbiology labs. Only one dye is utilized throughout the staining procedure in this form of staining. It's most commonly used to show bacterial shape and organization.
The dye stains the bacteria or its components.
Carbol fuchsin stain, for example.
Blue methylene stain Stain of crystal violet.
2. Differential staining method
In the differential staining approach, multiple stains are utilized to distinguish various cell shapes and/or cell types. Gram and Ziehl-Nelson stains, for example.
Gram staining was developed by a scientist called Christian Gram.
Because of changes in cell wall structure, most bacteria may be distinguished by their gram reaction. Gram-positive bacteria decolorize with acetone-alcohol and then stain purple with crystal violet. Gram-negative bacteria lose their primary stain (crystal violet) after being treated with acetone-alcohol and stain pink with the counter stain (safranin).
Required reagents:
. Gram’s Iodine
. Acetone-Alcohol
. Safranin
Procedure:
1. Prepare the smear from the culture or from the specimen.
2. Allow the smear to air-dry completely.
3. Rapidly pass the slide (smear upper most) three times through
the flame.
4. Cover the fixed smear with crystal violet for 1 minute and wash
with distilled water.
5. Tip off the water and cover the smear with gram’s iodine for 1
minute.
6. Wash off the iodine with clean water.
7. Decolorize rapidly with acetone-alcohol for 30 seconds.
8. Wash off the acetone-alcohol with clean water.
9. Cover the smear with safranin for 1 minute.
10. Wash off the stain wipe the back of the slide. Let the smear to
air-dry.
11. Examine the smear with oil immersion objective to look for
bacteria.
Interpretation of results:
.Gram-positive appears blue or purple after staining procedure.
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