Papanicolaou stain

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Papanicolaou stain showing a low-grade squamous intraepithelial lesion (LSIL) from a Pap test. Cell nuclei stained blue. Low-grade squamous intraepithelial lesion.jpg
Papanicolaou stain showing a low-grade squamous intraepithelial lesion (LSIL) from a Pap test. Cell nuclei stained blue.

Papanicolaou stain (also Papanicolaou's stain and Pap stain) is a multichromatic (multicolored) cytological staining technique developed by George Papanicolaou in 1942. [1] [2] [3] The Papanicolaou stain is one of the most widely used stains in cytology, [1] where it is used to aid pathologists in making a diagnosis. Although most notable for its use in the detection of cervical cancer in the Pap test or Pap smear, it is also used to stain non-gynecological specimen preparations from a variety of bodily secretions and from small needle biopsies of organs and tissues. [4] [5] Papanicolaou published three formulations of this stain in 1942, 1954, and 1960. [2]

Contents

Usage

Pap staining is used to differentiate cells in smear preparations (in which samples are spread or smeared onto a glass microscope slide) [6] from various bodily secretions and needle biopsies; the specimens may include gynecological smears (Pap smears), sputum, brushings, washings, urine, cerebrospinal fluid, [4] abdominal fluid, pleural fluid, synovial fluid, seminal fluid, [7] fine needle aspirations, tumor touch samples, or other materials containing loose cells. [8] [4] [9]

The pap stain is not fully standardized and comes in several formulations, differing in the exact dyes used, their ratios, and timing of the process. [2] [1] Pap staining is usually associated with cytopathology in which loose cells are examined, but the stain has also been modified and used on tissue slices. [9]

Pap test

Pap staining is used in the Pap smear (or Pap test) and is a reliable technique in cervical cancer screening in gynecology. [10]

Generalized staining method

The classic form of the Papanicolaou stain involves five stains in three solutions. [2] [11] [12]

The counterstains are dissolved in 95% ethyl alcohol which prevents cells from over staining which would obscure nuclear detail and cell outlines especially in the case when cells are overlapping on the slide. [3] [2] Phosphotungstic acid is added to adjust the pH of counterstains and helps to optimize the color intensity. [2] The EA counterstain contains Bismarck brown and phosphotungstic acid, which when in combination, cause both to precipitate out of solution, reducing the useful life of the mixture. [2]

Results

The stain should result in cells that are fairly transparent so even thicker specimens with overlapping cells can be interpreted. [2] Cell nuclei should be crisp, blue to black on color [12] [13] and the chromatin patterns of the nucleus should be well defined. Cell cytoplasm stains blue-green and keratin stains orange in color. [13] [5]

Eosin Y stains the superficial epithelial squamous cells, nucleoli, cilia, and red blood cells. [2] Light Green SF yellowish confers a blue staining for the cytoplasm of active cells such as columnar cells, parabasal squamous cells, and intermediate squamous cells. [14] Superficial cells are orange to pink, and intermediate and parabasal cells are turquoise green to blue. [12]

Ultrafast Papanicolaou stain

Ultrafast Papanicolaou stain is an alternative for the fine needle aspiration samples, developed to achieve comparable visual clarity in significantly shorter time. The process differs in rehydration of the air-dried smear with saline, use 4% formaldehyde in 65% ethanol fixative, and use of Richard-Allan Hematoxylin-2 and Cyto-Stain, resulting in a 90-second process yielding transparent polychromatic stains. [15]

Examples of Papanicolaou stain


Papers by George N. Papanicolaou describing his stain

See also

Related Research Articles

<span class="mw-page-title-main">Pap test</span> Cervical screening test to detect potential cancers

The Papanicolaou test is a method of cervical screening used to detect potentially precancerous and cancerous processes in the cervix or colon. Abnormal findings are often followed up by more sensitive diagnostic procedures and, if warranted, interventions that aim to prevent progression to cervical cancer. The test was independently invented in the 1920s by the Greek physician Georgios Papanikolaou and named after him. A simplified version of the test was introduced by the Canadian obstetrician Anna Marion Hilliard in 1957.

<span class="mw-page-title-main">Haematoxylin</span> Natural stain derived from hearthwood and used in histology

Haematoxylin or hematoxylin, also called natural black 1 or C.I. 75290, is a compound extracted from heartwood of the logwood tree with a chemical formula of C
16H
14O
6. This naturally derived dye has been used as a histologic stain, as an ink and as a dye in the textile and leather industry. As a dye, haematoxylin has been called palo de Campeche, logwood extract, bluewood and blackwood. In histology, haematoxylin staining is commonly followed by counterstaining with eosin. When paired, this staining procedure is known as H&E staining and is one of the most commonly used combinations in histology. In addition to its use in the H&E stain, haematoxylin is also a component of the Papanicolaou stain which is widely used in the study of cytology specimens.

<span class="mw-page-title-main">Georgios Papanikolaou</span> Greek pathologist (1883–1962)

Georgios Nikolaou Papanikolaou was a Greek physician, zoologist and microscopist who was a pioneer in cytopathology and early cancer detection, and inventor of the "Pap smear".

<span class="mw-page-title-main">Cytopathology</span> A branch of pathology that studies and diagnoses diseases on the cellular level

Cytopathology is a branch of pathology that studies and diagnoses diseases on the cellular level. The discipline was founded by George Nicolas Papanicolaou in 1928. Cytopathology is generally used on samples of free cells or tissue fragments, in contrast to histopathology, which studies whole tissues. Cytopathology is frequently, less precisely, called "cytology", which means "the study of cells".

<span class="mw-page-title-main">Romanowsky stain</span> Family of related stains for examination of blood including the detection of parasites

Romanowsky staining, also known as Romanowsky–Giemsa staining, is a prototypical staining technique that was the forerunner of several distinct but similar stains widely used in hematology and cytopathology. Romanowsky-type stains are used to differentiate cells for microscopic examination in pathological specimens, especially blood and bone marrow films, and to detect parasites such as malaria within the blood. Stains that are related to or derived from the Romanowsky-type stains include Giemsa, Jenner, Wright, Field, May–Grünwald and Leishman stains. The staining technique is named after the Russian physician Dmitri Leonidovich Romanowsky (1861–1921), who was one of the first to recognize its potential for use as a blood stain.

<span class="mw-page-title-main">Staining</span> Technique used to enhance visual contrast of specimens observed under a microscope

Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Stains and dyes are frequently used in histology, in cytology, and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses of diseases at the microscopic level. Stains may be used to define biological tissues, cell populations, or organelles within individual cells.

<span class="mw-page-title-main">Fine-needle aspiration</span> Diagnostic medical procedure

Fine-needle aspiration (FNA) is a diagnostic procedure used to investigate lumps or masses. In this technique, a thin, hollow needle is inserted into the mass for sampling of cells that, after being stained, are examined under a microscope (biopsy). The sampling and biopsy considered together are called fine-needle aspiration biopsy (FNAB) or fine-needle aspiration cytology (FNAC). Fine-needle aspiration biopsies are very safe minor surgical procedures. Often, a major surgical biopsy can be avoided by performing a needle aspiration biopsy instead, eliminating the need for hospitalization. In 1981, the first fine-needle aspiration biopsy in the United States was done at Maimonides Medical Center. Today, this procedure is widely used in the diagnosis of cancer and inflammatory conditions. Fine needle aspiration is generally considered a safe procedure. Complications are infrequent.

<span class="mw-page-title-main">Light green SF</span> Chemical compound

Light green SF, also called C.I. 42095, light green SF yellowish, is a green triarylmethane dye.

<span class="mw-page-title-main">Orange G</span> Chemical compound and histologic stain

Orange G also called C.I. 16230, Acid Orange 10, or orange gelb is a synthetic azo dye used in histology in many staining formulations. It usually comes as a disodium salt. It has the appearance of orange crystals or powder.

<span class="mw-page-title-main">Koilocyte</span>

A koilocyte is a squamous epithelial cell that has undergone a number of structural changes, which occur as a result of infection of the cell by human papillomavirus (HPV). Identification of these cells by pathologists can be useful in diagnosing various HPV-associated lesions.

Aurel A. Babeș was a Romanian scientist and one of the discoverers of the vaginal smear as screening test for cervical cancer.

<span class="mw-page-title-main">Diff-Quik</span>

Diff-Quik is a commercial Romanowsky stain variant used to rapidly stain and differentiate a variety of pathology specimens. It is most frequently used for blood films and cytopathological smears, including fine needle aspirates. The Diff-Quik procedure is based on a modification of the Wright-Giemsa stain pioneered by Harleco in the 1970s, and has advantages over the routine Wright-Giemsa staining technique in that it reduces the 4-minute process into a much shorter operation and allows for selective increased eosinophilic or basophilic staining depending upon the time the smear is left in the staining solutions.

The Bethesda system (TBS), officially called The Bethesda System for Reporting Cervical Cytology, is a system for reporting cervical or vaginal cytologic diagnoses, used for reporting Pap smear results. It was introduced in 1988 and revised in 1991, 2001, and 2014. The name comes from the location of the conference, sponsored by the National Institutes of Health, that established the system.

FNA mapping is an application of fine-needle aspiration (FNA) to the testis for the diagnosis of male infertility. FNA cytology has been used to examine pathological human tissue from various organs for over 100 years. As an alternative to open testicular biopsy for the last 40 years, FNA mapping has helped to characterize states of human male infertility due to defective spermatogenesis. Although recognized as a reliable, and informative technique, testis FNA has not been widely used in U.S. to evaluate male infertility. Recently, however, testicular FNA has gained popularity as both a diagnostic and therapeutic tool for the management of clinical male infertility for several reasons:

  1. The testis is an ideal organ for evaluation by FNA because of its uniform cellularity and easy accessibility.
  2. The trend toward minimally invasive procedures and cost-containment views FNA favorably compared to surgical testis biopsy.
  3. The realization that the specific histologic abnormality observed on testis biopsy has no definite correlation to either the etiology of infertility or to the ability to find sperm for assisted reproduction.
  4. Assisted reproduction has undergone dramatic advances such that testis sperm are routinely used for biological pregnancies, thus fueling the development of novel FNA techniques to both locate and procure sperm.
<span class="mw-page-title-main">Calcofluor-white</span> Fluorescent blue dye

Calcofluor-white or CFW is a fluorescent blue dye used in biology and textiles. It binds to 1-3 beta and 1-4 beta polysaccharides of chitin and cellulose that are present in cell walls on fungi, plants, and algae.

<span class="mw-page-title-main">Cervical screening</span> Type of medical screening

Cervical cancer screening is a medical screening test designed to identify risk of cervical cancer. Cervical screening may involve looking for viral DNA, and/or to identify abnormal, potentially precancerous cells within the cervix as well as cells that have progressed to early stages of cervical cancer. One goal of cervical screening is to allow for intervention and treatment so abnormal lesions can be removed prior to progression to cancer. An additional goal is to decrease mortality from cervical cancer by identifying cancerous lesions in their early stages and providing treatment prior to progression to more invasive disease.

<span class="mw-page-title-main">Liquid-based cytology</span> Method of preparing samples for cytopathology

Liquid-based cytology is a method of preparing samples for examination in cytopathology. The sample is collected, normally by a small brush, in the same way as for a conventional smear test, but rather than the smear being transferred directly to a microscope slide, the sample is deposited into a small bottle of preservative liquid. At the laboratory, the liquid is treated to remove other elements such as mucus before a layer of cells is placed on a slide.

<span class="mw-page-title-main">Sampling (medicine)</span> Collection of bodily substances for medical assessment

In medicine, sampling is gathering of matter from the body to aid in the process of a medical diagnosis and/or evaluation of an indication for treatment, further medical tests or other procedures. In this sense, the sample is the gathered matter, and the sampling tool or sampler is the person or material to collect the sample.

John Kingsbury Frost was an American physician specializing in the field of cytopathology - the microscopic study of individual body cells to detect cancer and other diseases. The first area of the body to be studied in this way was the female genital tract, using the Pap smear invented by Georgios Papanikolaou. Frost and other physicians expanded the field to allow for cytopathologic evaluation of the lung, bladder, and many other body sites. Frost was best known as a teacher of cytopathology. He organized and directed a school of cytotechnology and created and led a postgraduate Institute to teach the techniques to physicians.

Leopold George Koss was an American physician, pathologist, and professor at the Albert Einstein College of Medicine. He has been called "one of the founding fathers for the field of cytopathology".

References

  1. 1 2 3 Schulte EK (1991). "Standardization of biological dyes and stains: pitfalls and possibilities". Histochemistry. 95 (4): 319–328. doi:10.1007/BF00266958. PMID   1708749. S2CID   29628388.
  2. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Gill, Gary W. (2013). "Papanicolaou Stain". Cytopreparation. Essentials in Cytopathology. Vol. 12. pp. 143–189. doi:10.1007/978-1-4614-4933-1_10. ISBN   978-1-4614-4932-4. ISSN   1574-9053.
  3. 1 2 Chantziantoniou N, Donnelly AD, Mukherjee M, Boon ME, Austin RM (2017). "Inception and Development of the Papanicolaou Stain Method". Acta Cytol. 61 (4–5): 266–280. doi: 10.1159/000457827 . PMID   28384641.{{cite journal}}: CS1 maint: multiple names: authors list (link)
  4. 1 2 3 Kumar, Vinay; Abbas, Abul K.; Aster, Jon C. (2013). Robbins basic pathology (9th ed.). Elsevier/Saunders. p. 910. ISBN   978-1-4377-1781-5.
  5. 1 2 Drury, R. A. B.; Wallington, E. A. (1980). Carleton's Histological Technique (5th ed.). Oxford University Press. pp. 520p. ISBN   0-19-261310-3.
  6. ↑ Stedman's medical dictionary (27th ed.). Lippincott Williams & Wilkins. 2006. ISBN   978-0683400076.
  7. ↑ Lars Björndahl; David Mortimer; Christopher L. R. Barratt (1 April 2010). A Practical Guide to Basic Laboratory Andrology. Cambridge University Press. ISBN   978-1-139-48249-3.
  8. ↑ Hoda RS (2007). "Non-gynecologic cytology on liquid-based preparations: A morphologic review of facts and artifacts". Diagnostic Cytopathology. 35 (10): 621–34. doi:10.1002/dc.20698. PMID   17854077. S2CID   38797168.
  9. 1 2 Preethi, S.; Sivapathasundharam, B. (2014). "Will modified Papanicolaou stain be the new stain for keratin?". Journal of Histotechnology. 38 (1): 9–13. doi:10.1179/2046023614Y.0000000053. ISSN   0147-8885. S2CID   84486076.
  10. 1 2 Ross, Michael H.; Pawlina, Wojciech (2016). Histology : a text and atlas : with correlated cell and molecular biology (7th ed.). Wolters Kluwer. pp. 984p. ISBN   978-1451187427.
  11. ↑ Carson, Freida L; Hladik, Christa (2009). Histotechnology: A Self-Instructional Text (3 ed.). Hong Kong: American Society for Clinical Pathology Press. pp. 361–3363. ISBN   978-0-89189-581-7.
  12. 1 2 3 4 5 6 Bancroft, John; Stevens, Alan, eds. (1982). The Theory and Practice of Histological Techniques (2nd ed.). Longman Group Limited.
  13. 1 2 Dey, Pranab (2018). "Routine Staining in Cytology Laboratory". Basic and Advanced Laboratory Techniques in Histopathology and Cytology. pp. 133–138. doi:10.1007/978-981-10-8252-8_14. ISBN   978-981-10-8251-1.
  14. ↑ Faith Mokobi (2020-09-09). "Papanicolaou Staining (Pap Stain) For Pap Smear / Pap Test".
  15. ↑ Yang GC, Alvarez II (1995). "Ultrafast Papanicolaou stain. An alternative preparation for fine needle aspiration cytology". Acta Cytol. 39 (1): 55–60. PMID   7531380.
  16. ↑ Demay, Richard (2012). "Chapter 26: Stains". The art and science of cytopathology. Chicago, IL: Am Soc Clinical Pathology. p. 1505. ISBN   978-0-89189-644-9. OCLC   761848930.
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