What is the procedure of Giemsa stain?
Method
- Deparaffinize and rehydrate through graded alcohols to water.
- Rinse in pH 6.8 buffered distilled water.
- Stain in working Giemsa, overnight.
- Rinse in distilled water.
- Rinse in 0.5% aqueous acetic acid until section is pink.
- Wash in tap water.
- Blot until almost dry.
How do you prepare Giemsa for staining?
Recipe
- Dissolve 3.8g of Giemsa powder into 250ml of methanol.
- Heat the solution from step 1 to ~60oC.
- Slowly add in 250ml of glycerin to the solution from step 2.
- Filter the solution from step 3.
- The solution needs to stand a period of time prior to use.
How do you make a 10% Giemsa solution?
Therefore, 4.5 mL of Giemsa stock solution should be mixed with 40.5 mL of buffered water to prepare the required amount of 10% Giemsa working solution for staining 15 individual blood films.
What are the two methods for Giemsa staining?
The two methods for staining with Giemsa stain are the rapid (10% stain working solution) and the slow (3% stain working solution) methods. The rapid (10% stain working solution) method This is the commonest method for staining 1–15 slides at a time.
Why Giemsa stain is used?
Giemsa stain makes it possible to demonstrate the presence of microorganisms in all types of tissues. It can especially be used to detect the presence of Helicobacter pylori in the diagnosis of gastric ulcers or chronic gastritis.
How do you perform a Gram stain procedure?
Gram Staining – YouTube
Why glycerol is used in Giemsa stain?
In 1904 Giemsa published an essay on the staining procedure for flagellates, blood cells, and bacteria. Giemsa improved the Romanowsky stain (Eosin Y and Methylene Blue) by stabilizing this dye solution with glycerol. This allowed for reproducible staining of cells for microscopy purposes.
How do you dilute Giemsa?
Giemsa stain (3 ml) is diluted with buffered distilled water (100 ml) and is the stain of choice for demonstrating the presence of parasites in blood smears.
What is the purpose of Giemsa stain?
Giemsa stain is performed on paraffin sections. It is used to stain the blood cells of hematopoietic tissues. It can also be applied to all tissue sections in which the presence of microorganisms is suspected. Gram + and Gram Bacteria are not differentiated with this staining.
How long does Giemsa stain take?
45-60 minutes
3. Place slides into the working Giemsa stain (2.5%) for 45-60 minutes. 4. Remove thin smear slides and rinse by dipping 3-4 times in the Giemsa buffer.
What are the 4 steps of Gram staining?
The Gram staining process includes four basic steps, including:
- Applying a primary stain (crystal violet).
- Adding a mordant (Gram’s iodine).
- Rapid decolorization with ethanol, acetone or a mixture of both.
- Counterstaining with safranin.
What are the 2 types of Gram stain?
There are two main categories of bacterial infections: Gram-positive and Gram-negative. The categories are diagnosed based on the how the bacteria reacts to the Gram stain. A Gram stain is colored purple. When the stain combines with bacteria in a sample, the bacteria will either stay purple or turn pink or red.
What is the pH of Giemsa stain?
StainRITE® Wright-Giemsa Stain Phosphate Buffer pH 6.8.
How do you make 500ml of Giemsa stain?
Preparation of the Giemsa Stain Stock solution (500ml)
- Into 250ml of methanol, add 3.8g of Giemsa powder and dissolve.
- Heat the solution up to ~60oC.
- Then, add 250ml of glycerin to the solution, slowly.
- Filter the solution and leave it to stand for about 1-2 months before use.
What type of stain is Giemsa?
Giemsa stain (/ˈɡiːmzə/), named after German chemist and bacteriologist Gustav Giemsa, is a nucleic acid stain used in cytogenetics and for the histopathological diagnosis of malaria and other parasites.
What bacteria is detected using Giemsa stain?
Giemsa stains the fungus Histoplasma, Chlamydia bacteria, and can be used to identify Mast cells.
What is the correct sequence for a Gram stain?
The stains are applied to a smear of bacteria on a microscope slide in the following order: crystal violet, Gram’s iodine, decolorizing agent, and safranin.
What are the 4 steps of gram staining?
What color is Gram-positive?
A Gram stain is colored purple. When the stain combines with bacteria in a sample, the bacteria will either stay purple or turn pink or red. If the bacteria stays purple, they are Gram-positive. If the bacteria turns pink or red, they are Gram-negative.
Is Giemsa stain acidic or basic?
Giemsa stain is a differential stain. It is composed of eosin and azure, both of which are acidic dyes, and methylene blue, which is a basic dye. The acidic dyes eosin and azure stain the basic cellular components such as cytoplasm and granules among others.
Which is applied first in Gram staining?
crystal violet dye
The first step in gram staining is the use of crystal violet dye for the slide’s initial staining. The next step, also known as fixing the dye, involves using iodine to form crystal violet- iodine complex to prevent easy removal of dye.