What stain is used for Plasmodium falciparum?
Abstract. The fluorescent dye benzothiocarboxypurine (BCP) intensely stains nucleic acids. The dye does not penetrate viable white blood cells but does stain these cells following fixation. It has also been found that the dye stains the nucleic acid of viable Plasmodium falciparum.
Why is Giemsa stain used in malaria?
Use of Giemsa stain is the recommended and most reliable procedure for staining thick and thin blood films. Giemsa solution is composed of eosin and methylene blue (azure). The eosin component stains the parasite nucleus red, while the methylene blue component stains the cytoplasm blue.
Which stain is used for detection of Plasmodium?
Giemsa stain
Giemsa stain – Recommended for detection and identification of blood parasites.
What is Giemsa stain used for?
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 do you identify Plasmodium falciparum?
Malaria parasites can be identified by examining under the microscope a drop of the patient’s blood, spread out as a “blood smear” on a microscope slide. Prior to examination, the specimen is stained (most often with the Giemsa stain) to give the parasites a distinctive appearance.
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 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 is Giemsa staining done?
Giemsa’s solution is a mixture of methylene blue, eosin, and Azure B. The stain is usually prepared from commercially available Giemsa powder. A thin film of the specimen on a microscope slide is fixed in pure methanol for 30 seconds, by immersing it or by putting a few drops of methanol on the slide.
How do you make a 10% Giemsa stain?
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.
Why is Giemsa stain preferred?
The Wright-Giemsa stain is the most commonly used Classical stain by technicians because of its versatility and consistent staining appearance. It is formulated to produce more intense basophilic/nuclear staining in blood cell morphology.
What Colour is Giemsa stain?
dark blue
Giemsa stain is one of the best known histological stains, coloring the nuclei dark blue and the cytoplasm blue to pink, according to the acidity of the cytoplasmic contents.
How do you differentiate P. falciparum and P vivax?
The main difference between Plasmodium falciparum and Plasmodium vivax is that P. falciparum causes severe malaria as it rapidly multiplies in the blood whereas P. vivax is less virulent than P.
What are the characteristics of Plasmodium falciparum?
Unlike those of other Plasmodium species, the gametocytes of P. falciparum are elongated and crescent-shaped, by which they are sometimes identified. A mature gametocyte is 8–12 μm long and 3–6 μm wide. The ookinete is also elongated measuring about 18–24 μm.
How do you make a 5% Giemsa stain solution?
Stock Solution
- 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.
What type of stain is Giemsa stain?
Giemsa stain is a buffered thiazine-eosinate solution designed to provide coloration of blood cells similar to the original product described by Giemsa. It may be used separately or in combination with a May Grünwald Stain, also available from Sigma-Aldrich.
What type of dye is Giemsa stain?
Giemsa stain is a differential stain and contains a mixture of azure, methylene blue, and eosin dye. It is specific for the phosphate groups of DNA and attaches itself to where there are high amounts of adenine-thymine bonding.
How do you make a 5% Giemsa?
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 does P. falciparum look like?
P. falciparum gametocytes are crescent or sausage shaped. The chromatin is in a single mass (macrogamete) or diffuse (microgamete). Gametocytes in a thick blood smear.
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.
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.
How do you make a 10% Giemsa?
How do you identify P. falciparum?
The most definitive finding of P. falciparum is the shape of the gametocytes. Unlike what we see in the other species of malaria, they are crescent-shaped or banana-shaped.