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.
Why is Giemsa referred to as a differential stain?
Giemsa stain is also a differential stain, such as when it is combined with Wright stain to form Wright-Giemsa stain. It can be used to study the adherence of pathogenic bacteria to human cells. It differentially stains human and bacterial cells purple and pink respectively.
What is the principle of Giemsa?
PRINCIPLE: The “neutral” dyes combining the basic dye methylene blue and the acid dye eosin, give a wide color range when staining. The pH of the staining solution is critical and ideally should be adjusted for different fixatives.
What are the steps of gram staining?
The performance of the Gram Stain on any sample requires 4 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 …
Who Giemsa stain preparation?
Giemsa stain preparation
- Dissolve 3.8g of Giemsa powder within 250ml of methanol.
- Heat the solution at 60oC.
- Add 250ml of glycerin.
- Filter the solution.
- leave the solution to stand for about 1-2 months before use. Store the solution in a cool, dark place.
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 differential staining technique?
Differential staining is a staining process which uses more than one chemical stain. Using multiple stains can better differentiate between different microorganisms or structures/cellular components of a single organism.
How do you distinguish between gram-positive and Gram negative bacteria?
Gram positive bacteria have a thick peptidoglycan layer and no outer lipid membrane whilst Gram negative bacteria have a thin peptidoglycan layer and have an outer lipid membrane.
What is the purpose of using a Giemsa staining technique on chromosomes?
G-banding, G banding or Giemsa banding is a technique used in cytogenetics to produce a visible karyotype by staining condensed chromosomes. It is useful for identifying genetic diseases through the photographic representation of the entire chromosome complement.
How do you dilute Giemsa?
Dilute Giemsa Stain 1:20 with deionized water. Color can be varied by diluting in buffer. 4. Stain film for 15-60 minutes.
How do you distinguish between Gram-positive and Gram-negative bacteria?
What are the 3 types of differential staining?
Article Summary: The Gram, Ziehl Neelsen acid fast, and endospore stains are differential tests used to help identify bacteria.
What is the purpose of a differential stain?
Differential staining is a procedure that takes advantage of differences in the physical and chemical properties of different groups of bacteria. It allows us to differentiate between different kinds of bacterial cells or different parts of a bacterial cell.
Why is it important to know if bacteria is Gram-positive or negative?
A Gram stain is a common laboratory test that can help diagnose the presence of a bacterial infection quickly. Healthcare providers often order it alongside a bacteria culture to diagnose the type of bacterium causing the infection.
Which stain is usually used to Colour chromosomes?
The stains commonly used for staining nucleic acid are Acetocarmine and Giemsa.
What nucleotides does the Giemsa dye stain?
Chromosomes are visualized using Giemsa staining (G-banding). Light bands represent early replicating regions, rich in guanine and cytosine nucleotides. Dark bands represent late replicating regions, rich in adenine and thymine nucleotides.
What is the function of glycerol in Giemsa stain?
In the early 1900s, Gustav Giemsa designed the Giemsa stain to detect parasites such as malaria and Treponema pallidum in blood smears. He developed a “secret” oxidation process using a unique mixture of methylene azure, methylene blue, and eosin, with glycerol added as a stabilizing agent.
How do you dilute Giemsa for malaria?
Stock Giemsa Buffer 10.0 ml Deionized water 990.0 ml Check pH before use. Should be 7.2. Stable at room temperature for one month. Deionized water (warmed to 56°C) 95.0 ml Triton X-100 5.0 ml Prewarm the deionized water and slowly add the Triton X-100, swirling to mix.
What are differential staining techniques?
What are the advantages of differential stain?
Differential staining can help identify a culture with more than one type of bacteria or other microorganism and also its physicalcharacteristics through gram staining, which can identify the makeup of the bacteria’s cell wall.