OG6 is a counter stain designed to stain the cytoplasm of cells derived from the human body. The stained cell sample is used to facilitate the diagnosis of the pathological state, and is intended for in-vitro examination of cellular deformity. PRINCIPLE OF METHOD: Papanicolaou staining (Pap.
What is the color of OG6 solution?
Papanicolaou’s solution 2a Orange G solution (OG6) gives a pale, yellow-orange staining result with mature and keratinized squamous cells.
How do you prepare OG6?
1) Rinse in 70% alcohol, 50% alcohol and distilled water. 2) Stain in Harris haematoxylin (without acetic acid) for 5 -10 minutes. 3) Rinse in distilled water. 4) Rinse 3 or 4 times in 0.5% aqueous solution of hydrochloric acid.
What is the next reagent used after OG6 in Paps staining procedure?
6. Post-OG and post-EA alcohol is replaced by 0.5% of acetic acid. Acid pH keeps acid dyes in cells as 95% alcohol. In addition, dilute acetic acid solution is too weak to remove hematoxylin as HCl used to differentiate in regressive procedure.
What does OG6 stain? – Related Questions
What is the most commonly used fixative in Pap smear?
Various fixatives are used in exfoliative cytology. Out of which, 95% ethanol is the commonly used fixative. Hence, the conventional method for fixation of Pap smear is to fix the Pap smear immediately in 95% ethyl alcohol after preparing the smear.
Which stain is used for Pap smear?
The PAP stain is a polychromatic counterstaining method consisting of stains such as Orange G 6 (OG6) and modified eosin azure (EA). The strength of the Pap stain is such that it results in: Well-stained nuclear chromatin. Differential polychromatic counterstaining of cytoplasm.
What is the correct order of staining reagents in Gram staining?
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.
Which is the correct order of solutions used during 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
What are the solutions reagents in order used in Gram staining?
Reagents needed for Gram staining include:
Crystal violet (primary stain) [1] Gram’s iodine solution (the mordant) [1] Acetone/ethanol (50:50 v:v) (the decolorizer) [1] 0.1% basic fuchsin solution (the counterstain) [1]
What is the fourth and final reagent used in the Gram stain?
The final step in gram staining is to use basic fuchsin stain to give decolorized gram-negative bacteria pink color for easier identification. It is also known as counterstain. Some laboratories use safranin as a counterstain; however, basic fuchsin stains gram-negative organisms more intensely than safranin.
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.
Why is Gram-positive purple?
Gram positive bacteria stain violet due to the presence of a thick layer of peptidoglycan in their cell walls, which retains the crystal violet these cells are stained with.
Is E coli pink or purple?
After gram staining, gram-positive Staphylococcus aureus turn purple and gram-negative Escherichia coli turn pink.
Why is Gram-negative pink?
Gram negative bacteria appear a pale reddish color when observed under a light microscope following Gram staining. This is because the structure of their cell wall is unable to retain the crystal violet stain so are colored only by the safranin counterstain.
How do you remember a gram positive purple?
Gram positive bacteria have a thick peptidoglycan cell wall that stains with crystal violet stain.
Use the mnemonic “CLAP”:
- C – Clostridium.
- L – Lactobacillus.
- A – Actinomyces.
- P – Propionibacterium.
Why is gram negative pink and Gram positive purple?
The Gram positive cell looses some of its large chunky peptidoglycan cell wall but keeps enough of it to retain the purple colour. The Gram negative cell has its outer membrane and small peptidoglycan layer completely stripped away, leaving it colourless.
Is purple stain Gram positive or negative?
gram stain test
Gram-positive bacteria remain purple because they have a single thick cell wall that is not easily penetrated by the solvent; gram-negative bacteria, however, are decolorized because they have cell walls with much thinner layers that allow removal of the dye by the solvent.
