What is the procedure of Giemsa stain?
For Thin blood smear Stain with diluted Giemsa stain (1:20, vol/vol) for 20 min (For a 1:20 dilution, add 2 ml of stock Giemsa to 40 ml of buffered water in a Coplin jar). Wash by briefly dipping the slide in and out of a Coplin jar of buffered water (one or two dips). Note: Excessive washing will decolorize the film.
What is the purpose of Giemsa stain?
Giemsa is the prototypical stain used to detect malaria and Trypanosoma-infected blood (Figure 5). Plasmodium falciparum gametocytes and mature trophozoites can be detected using thin and thick smears, respectively. WBCs, platelets, and remnants of RBCs are also visible with Giemsa staining on thin and thick smears.
What stain is used to stain blood smear?
Blood films are routinely stained with a Romanowsky-type stain (e.g., Wright or Wright-Giemsa) either manually or using an automatic slide stainer. Romanowsky-type stains are composed of a mixture of eosin and oxidized methylene blue (azure) dyes.
What are the steps of Gram staining?
The performance of the Gram Stain on any sample requires four basic steps that include applying a primary stain (crystal violet) to a heat-fixed smear, followed by the addition of a mordant (Gram’s Iodine), rapid decolorization with alcohol, acetone, or a mixture of alcohol and acetone and lastly, counterstaining with …
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.
What is the size of RBC?
about 7-8 µm
Normal human RBCs have a biconcave shape, their diameter is about 7-8 µm, and their thickness is about 2.5 µm [11,12,34].
What is Field stain A and B?
Field stain is a histological method for staining of blood smears. It is used for staining thick blood films in order to discover malarial parasites. Field’s stain consists of two parts – Field’s stain A is methylene blue and Azure 1 dissolved in phosphate buffer solution; Field’s stain B is Eosin Y in buffer solution.
What is the principle behind the Gram stain reaction?
The basic principle of gram staining involves the ability of the bacterial cell wall to retain the crystal violet dye during solvent treatment. Gram-positive microorganisms have higher peptidoglycan content, whereas gram-negative organisms have higher lipid content.
What is the most important step in any staining procedure?
Poor staining technique could lead to inaccurate results. One of the most important steps in Gram staining is the decolorizing step (use of alcohol/acetone). If the decolorizer is not left on long enough, then it will not be able to differentiate between Gram positive and Gram negative bacteria.
What is the most important step in the Gram staining procedure Why?
The thickness of the smear used in the Gram stain will affect the result of the stain. The step that is most crucial in effecting the outcome of the stain is the decolorizing step.
How do you prepare a solution of Giemsa stain?
Make up a 10% Giemsa solution with distilled/deionized water buffered to pH 7.2. If only one slide is to be stained, you will require about 3 ml of prepared stain. Allow 3 drops of stock Giemsa solution (from the Pasteur pipette) to each millilitre of buffered water to give a 10% solution.
What’s the procedure for making a Giemsa stain?
Add a thick smear of blood and air dry for 1 hour on a staining rack. Dip the thick blood smear into diluted Giemsa stain (prepared by taking 1ml of the stock solution and adding to 49ml of phosphate buffer or distilled water, but the results may vary differently).
How to make a thin film of Giemsa powder?
Into 250ml of methanol, add 3.8g of Giemsa powder and dissolve. Then, add 250ml of glycerin to the solution, slowly. Filter the solution and leave it to stand for about 1-2 months before use. On a clean dry microscopic glass slide, make a thin film of the specimen (blood) and leave to air dry.
What’s the procedure for staining a blood smear?
Procedure for staining • Pour Leishman’s stain dropwise (counting the drops) on the slide and wait for 2 minutes. This allows fixation of the PBF in methyl alcohol. • Add double the quantity of buffered water dropwise over the slide (i.e. double the number of drops).
What’s the best way to use Leishman’s stain?
Staining method with Leishman’s stain. 1) Prepare smear and air dry it. 2) Cover the smear with stain and wait for 2 mints for fixation. 3) Dilute with double volume of buffer water and wait for 7- 10 minutes for staining.