What is the staining procedure of Pap smear?

What is the staining procedure of Pap smear?

Pap staining procedure In regressive method the smear is over stained with nuclear stain with a non-acidified hematoxylin. The excess stain is removed with dilute HCL acid solution. Harris hematoxylin is preferred for regressive method. The progressive method is the most commonly used method in laboratories today.

What fixation is used for Pap cytological staining?

Rapid fixation in alcohol (wet version) is essential for pap staining, which brings out nuclear details clearly, allowing better identification of malignant cells.

What is rapid Pap stain?

The traditional Pap stain involves wet fixation and subsequent staining, together requiring at least 30 minutes. To cut down the time, the rapid Pap stains were developed by Kline,[1] Tao[3] and Sato[7] with respective staining time of 4 minutes, 5 minutes and 90 seconds.

What is the most critical step in the PAP staining?

As a limitation of this study, we should mention that UF-PAP stain is very sensitive technique, thus air-drying is a critical step. Also, due to the omission of O-G-6 in UF-PAP method, this method cannot be used for the diagnosis of squamous cell carcinoma.

What are the types of fixatives?

Popular fixative solutions

  • Phosphate buffered formalin.
  • Formal calcium.
  • Formal saline.
  • Zinc formalin (unbuffered)
  • Zenker’s fixative.
  • Helly’s fixative.
  • B-5 fixative.
  • Bouin’s solution.

What fixative is used for Pap smear?

A 95% solution of ethyl alcohol or spray fixative is used to fix the slide. The slides should be labeled in pencil with the patient’s name AND date of birth and a properly completed Pap smear requisition should accompany the slide.

Why Pap stain is used?

Pap stain is a universal stain used for gynecologic and non-gynecologic cytology smear. It is mainly used for oral and cervical cancer screening in asymptomatic population and in the follow up of patients with cancer. Pap test has decreased incidence of cervical cancer by 70% in developed countries.

How do you use H and E stain?

In the following sections, the basic steps in performing an H&E stain are outlined.

  1. Remove the Wax.
  2. Hydrate the Section.
  3. Apply the Hematoxylin Nuclear Stain.
  4. Complete the Nuclear Stain by “Blueing”
  5. Remove Excess Background Stain (Differentiate)
  6. Apply the Eosin Counterstain.

What is the principle of H and E staining?

The principle behind H & E stain is the chemical attraction between tissue and dye. Hematoxylin, a basic dye imparts blue-purple contrast on basophilic structures, primarily those containing nucleic acid moeties such as chromtatin, ribosomes and cytoplasmic regions rich in RNA.

What are the three types of fixatives?

What are the reagents used in Pap stain how is it done?

EA-50, or Modified EA-50, or EA-65 stain for 2.5 minutes. 95% Ethanol 10 dips, 2 changes. 100% Ethanol 1 minute. Clear in 2 changes of xylene, 2 minutes each.

Which is cytological fixative?

Abstract. Ninety-five percent (95%) ethanol is the standard cytological fixative used in many laboratories. Commercially available ethanol is expensive and not freely available in some institutions. Methanol, a tissue dehydrant, is also known to be a cytological fixative.

What is the principle of H and E?

Why do we use H and E?

Introduction. The H&E stain provides a comprehensive picture of the microanatomy of organs and tissues. Hematoxylin precisely stains nuclear components, including heterochromatin and nucleoli, while eosin stains cytoplasmic components including collagen and elastic fibers, muscle fibers and red blood cells.

What are the two types of fixative?

Classification of Fixative:

  • Aldehyde Fixatives: Formaldehyde Fixatives. Gluteraldehyde Fixatives.
  • Oxidizing Agents: Osmium tetroxide. Potassium permanganate.
  • Protein Denaturing Agents: Acetic Acid. Methyl Alcohol.
  • Cross Linking Agents: Carbodiimides. Dimethyl Subrimidate.
  • Unknown Mechanism/Miscellaneous: Mercuric Chloride.

What are fixatives?

Definition of fixative : something that fixes or sets: such as. a : a substance added to a perfume especially to prevent too rapid evaporation. b : a substance used to fix living tissue.

What are the techniques used in staining of cells in cytology?

Two of the most commonly used cytological stains are: • Papanicolaou (PAP) stain – Papanicolaou stain is a modi- fied hematoxylin and eosin (H&E) stain and is recom- mended for the staining of alcohol fixed cytology slides 1.

  • August 17, 2022