Papanicolaou staining technique pdf

The papanicolaou test abbreviated as pap test, also known as pap smear, cervical smear, cervical screening or smear test is a method of cervical screening used to detect potentially precancerous and cancerous processes in the cervix opening of the uterus or womb or colon in both women and men. This stain reveals nuclear detailsmembrane irregularities, chromatin pattern, and intranuclear invaginationsimportant for establishing a diagnosis of malignancy, especially welldifferentiated neoplasms that lack significant. The universal stain for oral cytological screening for precancer and cancer of oral cavity is papanicolaou stain which has been used in different. Pdf comparing triomodified papanicolaou staining methods. Papanicolaou stain, for in vitro diagnostic use sigmaaldrich. The technique uses a high number of dyes in its procedure. Papanicolaou stain definition of papanicolaou stain by. Comparison of modified ultrafast papanicolaou stain with. Pap staining procedure is classified into two types i.

Mast cells and basophils are readily recognized with a romanowsky stain, but they are not recognizable in papanicolaou stained smears or in cell blocks stained with hematoxylin and eosin. This staining technique divides bacteria into two groups namely acidfast and non acidfast. Papanicolaou smear cytology is widely accepted as a valuable diagnostic tool in the detection of cervical cancer and has also been described to be useful in the diagnosis of external ocular tumors. Cyto prep techniques questions and study guide quizlet. To study the efficacy of the ultrafast papanicolaou ufp staining technique for immediate cytologic diagnosis, to check specimen adequacy during radiologically guided fine needle aspiration. Nnewer techniquesewer techniques a conventional pap smear has a sensitivity ranging. The schaefferfulton method the most commonly used endospore staining technique uses heat to push the primary stain malachite green into the endospore. Shandon rapidchrome papanicolaou staining kit, papanicolaou. Leishman giemsa lg cocktail, being a relatively new staining technique, has not been used in exfoliative cytology. It also eliminates several steps to save time and expense.

It is a polychromatic stain containing multiple dyes to differentially stain various components of the cells. Washing with water decolorizes the cell, but the endospore retains the green stain. Pdf modified ultrafast papanicolaou staining technique. Sep, 2012 1865bohmer adds mordant to hematoxylin to make first successful hematoxylin stain 1. Guidelines of the papanicolaou society of cytopathology for. Papanicolaou stain for cytological preparations newcomer supply. The study is designed to search for a superior and improved qualitative staining technique which is. The first, between 1933 and 1942, saw the introduction of alcoholether fixation and aqueous waterblue staining to enhance cellular transparency, aiding the distinction of cervical cancer cells from benign cells, with quantitative and qualitative assessment of squamous cell maturity. Papanicolaou test definition of papanicolaou test by.

It discusses the principles of and offers clear guidance on all routine and special laboratory techniques. Revised february 2016 papanicolaou stain for cytological. Of these ultrafast pap stain ufp has become popular. Comparison of ultrafast papanicolaou stain with the.

A papanicolaou stain is typically performed in the laboratory on alcoholfixed slides. Approved guidelinethird edition this document discusses procedures for cervicovaginal specimen collection, as well as the preparation, fixation, staining, and storage of papanicolaoustained cervicovaginal cytology slides. This technique was further modified in india as many of the reagents. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou, the father of cytopathology. To standardize an inexpensive and rapid papanicolaou staining technique with limited ethanol usage. Staining was performed by a rapid manual technique stain is commonly used when there is need to examine the blood smear for the parasites but is a good stain for routine examination of blood smear and used to differentiate nuclear and cytoplasmic morphology of the various cells of the blood like platelets, rbcs giemsa staining technique. Comparison of ultrafast papanicolaou stain with the standard papanicolaou stain in body fluids and fine needle aspiration specimens. Wellstained cells are transparent, allowing the observer to interpret any underlying cells. Proper technique procedure, preparation of smears, fixation, staining. A greater knowledge of how to optimize or modify the technique to achieve the results desired a greater knowledge of how to solve staining issues. Stain in papanicolaou stain, ea36, ea50 or ea65 of choice for minutes, depending on specimen type and preference of stain intensity. Cytometry 7101103 1986 technical note a protocol for papanicolaou staining of cytologic specimens following flow analysis todd k. Papanicolaou stain papanikolow, a multichromatic stain used principally on exfoliated cytologic specimens and based on aqueous hematoxylin with multiple counterstaining dyes in 95% ethyl alcohol, giving great transparency and delicacy of detail.

The universal stain for cervical cytological screening is papanicolaou stain which has been used in different laboratories with many modifications. In the present study, a modified staining technique was adapted where gills hematoxylin was replaced by harris hematoxylin. The 5dye pap stain method evolved through 2 salient phases. Papanicolaou society of cytopathology fineneedle aspiration fna is a simple, safe, and costeffective procedure for the investigation of patients with a mass. M r srinivasan, a sabarinathan, a geetha, k shalini, m sowmiya. Pap smear staining papanicolaou staining technique. This procedure is extensively used in the diagnosis of tuberculosis and leprosy. The choice of which green dye to use will depend upon staining intensity preference. The specimen and staining solutions were sequen tially added to a 5micrometer pore size, 47 mm diameter gelman metricel filter while. The specimen pre staining fixative solution is a 10%. A protocol for papanicolaou staining of cytologic specimens. Pap test pap smear vaginal or cervical cells obtained for cytological study.

The study revealed that the papanicolaou staining is a better method over crystal violet staining technique, because the papanicolaou staining is polychromatic, hence it yields clear nuclear and cytoplasmic details of vaginal exfoliativecells. Ultrafast papanicolaou stain ufp was introduced as a hybrid of romanowsky and papanicolaou pap stain. Comparison of three staining methods for the morphological. Acosta the evaluation of sperm morphology is still an important parameter in the diagnosis of the infertile male. Mycobacterium tuberculosis is the most important of this group, as it is responsible for the disease called tuberculosis tb along. This technique was developed by george papanicolaou, the father of cytopathology. Basic and advanced laboratory techniques in histopathology. The dye stains the background but does not penetrate the capsules, which appear like halos around the borders of the cell. The papanicolaou stain solutions can be used for either manual or automated staining. Cell staining is a technique used for the main purpose of increasing contrast through changing the color of some of the parts of the structure being observed thus allowing for a clearer view. The pap stain, ea also varies with each ea formulation made with either light green sf or modified with fast green. This method enhances the nuclear component, increases cytoplasmic transparency, and reduces background staining. Background the present study was designed to determine whether the thinprep plus papanicolaou stain thinprep method is more sensitive than the cytospincoupled wrightgiemsa wg stain cytospin method in diagnosis of leptomeningeal metastasis lm from malignant solid tumors in cerebrospinal fluid csf.

Cell staining in microscopy types, techniques, preparations. Papanicolaou, papanicolaou stain underwent various modifications. Evolution of pap stain biomedical research and therapy. The science and application of hematoxylin and eosin staining. They can be recognized in toluidine bluestained wet films fig.

In addition, it covers various advanced laboratory techniques, such as immunocytochemistry, flow cytometry, liquid based cytology, polymerase chain reaction, tissue microarray, and molecular technology. We also explored if the thinprep method could be used in the differential diagnosis of. Sep 21, 2015 rehydration of air dried smears unfixed, airdried gynaecological smears received from peripheral areas can be used for papanicolaou staining by rehydration method the simplest rehydration technique is to place air dried cytological specimens 50 % aqueous solution of glycerine 2 rinses in 95% ethyl alcohol pap staining 44. Inception and development of the papanicolaou stain method.

The specimen does not need to be heatfixed prior to negative staining. This stain is a frequently used method for examining cell samples from a variety of tissue types in various organs. Papanicolaou stain pap stain protocol ihc world, llc. Over oxidation by excessive air contact over time will cause staining problems. Papanicolaou 18831962 that malignant uterine tumors slough off cancerous cells into surrounding vaginal fluid. The technique involves papanicolaou stain ing of cells on a membrane filter that has been previously stained with acridine orange and fixed with glutaraldehyde buffer. Papanicolaou staining is applied to vaginal exudates for the detection of uterine or vaginal cancer. Over the years cytopathologist have felt the need for a rapid pap stain that is as fast as diff quik and provides cytomorphological features as exquisite as does pap stain. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic multicolored cytological staining technique developed by george papanicolaou in 1942. Rehydration of air dried smears unfixed, airdried gynaecological smears received from peripheral areas can be used for papanicolaou staining by rehydration method the simplest rehydration technique is to place air dried cytological specimens 50 % aqueous solution of glycerine 2 rinses in 95% ethyl alcohol pap staining 44. The technique involves papanicolaou stain ing of cells on a membrane filter that has been previously stained with. Papanicolaou staining is commonly used for staining exfoliative cytology smears with romanowsky stains being used sparingly. Comparison of the routine papanicolaou staining technique with the rapid, economic, acetic acid, papanicolaou reap technique asthana 2a1, singh ak abstract background. Achieve rapid results of fine needle aspirations or highrisk specimens with the thermo scientific shandon rapidchrome papanicolaou staining kit.

A staining technique using modified mayers hematoxylin and supersaturated lithium carbonate for aspiration cytology and body fluid smear is described. The principle of pap stain is to clearly distinguish between basophilic and. The aim of this study was to investigate the possible application of ultrafast pap ufpap method in cytology. New and reproducible modified papanicolaou staining procedure. An average pap smear slide contains 50,000300,000 cells that must be examined and if the sample contains only a few abnormal cells within a crowded background of healthy cells, the abnormal cells may be missed. Refers to the improved papanicolaou staining method. Papanicolaou stain an overview sciencedirect topics.

Modified ultrafast papanicolaou staining technique. Two slides were immersed in 95% ethanol for staining with standard pap stain. On the basis of these results, the 1992 who manual 16 introduced the diffquik method together with a number of other staining. Pap staining is used to differentiate cells in smear preparations of various bodily secretions. Method details for use in the papanicolaou technique are given below. Papanicolaou stain imparts a characteristic range of coloration to exfoliative cells of vaginal, cervical, prostatic and other body secretions, allowing critical examination of nuclei and cytoplasmic components. Stain in papanicolaou stain, og6 for 30 seconds to 2 minutes, depending on specimen type and preference of stain intensity. A method for staining a pap test slide specimen for conventional pap test and, particularly a thinprep pap test specimen, in which a number of multiple hydrating, staining and dehydrating step are performed in an equal number of 10 to 15 dippings at about equal dip times of about 1 second. This item has been discontinued and is no longer available. Approved guidelinethird edition this document discusses procedures for cervicovaginal specimen collection, as well as the preparation, fixation, staining, and storage of papanicolaou stained cervicovaginal cytology slides. Mufp stain in comparison to routine pap, h and e, and giemsa provides an excellent and suitable alterative in cytological staining for the study of various organs. Most techniques used for staining human sperm are very timeconsuming. Journal of haracolog linical research how to cite this article. Singh et al pap staining for sperm morphologic analysis features in smears.

Diagnostic implications for routine diagnostic cytology to aid in the identification and classification of exfoliative cells. Microscopy refers to the practice that involves the use of a microscope for the purposes of observing small scale structures that cannot be viewed using the naked eye and often cell staining is necessary as s tructures are difficult to discern due to insufficient contrast. Airdried material normally submitted for diffquik modified romanowsky stain was rehydrated in normal saline, then fixed for a short period in formol alcohol, before staining by a modified papanicolaou technique. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou. Papanicolaou stain also papanicolaou s stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou, the father of cytopathology. Stains and dyes are frequently used in histology the study of tissue under the microscope and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses disease at a microscopic level. Papanicolaou stains are used to differentiate cells in smear preparations of various body secretions. Staining is a technique used to enhance contrast in samples, generally at the microscopic level. Either a progressive or regressive technique may be. The modified technique is referred as rapid, economic, acetic acid papanicolaou stain reap. A comparative study on staining techniques for vaginal. Dec 12, 2018 papanicolaou stains are used to differentiate cells in smear preparations of various body secretions.

Malignant cells are generally characterized by relatively large nuclei with an abnormal chromatin pattern and small amounts of cytoplasm. Importance of papanicolaou staining for sperm morphologic. In the present study, although there was a good correlation between the methods, cases in which there was no agreement of results had more midpiece and tail defects falsenegative or a lower value by the sqaiicp system owing to low smi values falsepositive. Us6284543b1 rapid papanicolaou staining method for. Papanicolaou stain a multichromatic stain used in cancer screening, especially of gynecologic smears. Smears from 200 patients were collected 2 per patient and fixed in. The papanicolaou stain is one of the most widely used stains in cytology, where it is used to aid pathologists in making a diagnosis. Papanicolaou stains ea50 s1060 and og6 s1070 and s0500 haematoxylin and cytopack ii sp800. Most cytology laboratories in all gulf countries including oman, use the standard papanicolaou pap method to stain various cytological specimens. Endospore staining uses two stains to differentiate endospores from the rest of the cell.

Papanicolaou staining, or pap staining, was developed to replace fine needle aspiration cytology fnac in hopes of decreasing staining times and cost without compromising quality. Pap staining is used to differentiate cells in smear preparations. Residents ppap smearap smear bioline international. The terms papanicolaou staining, papanicolaou staining, papanicolaou test, etc. Comparison of ultrafast papanicolaou stain with the standard.

New and reproducible modified papanicolaou staining. Papanicolaou stain is a multichromatic multicolored cytological staining technique developed. Thinprep plus papanicolaou stain method is more sensitive. Harris hematoxylin is the optimum nuclear stain and the combination of og6 and ea50 give the subtle range of green, blue and pink hues to the cell cytoplasm. Papanicolaou stain purpose of stain a polychromatic cytological staining technique used to differentiate cells in smear preparations of various bodily secretions. To optimize quantitation of dna on routine cytolog ical material, a technique for destaining papanicolaou stained slides and restaining them by the feulgen. The outcome of this method is crisp nuclear detail and transparency of the cytoplasm, which allows the examiner to clearly visualize cellular morphology. There are a variety of microscopic stains that can be used in microscopy. A quick, reliable staining technique for human sperm.

Cell staining in microscopy types, techniques, preparations and procedures. The hematoxylin and eosin stain a better understanding of the basic chemistry of the h and e stain will provide the attendee with. Theory and practice of histological techniques, edited by bancroft jd and gamble, m, churchill livingstone, new york, 2002, p129. Jan 12, 2020 papanicolaou stain also papanicolaous stain or pap stain is the most important stain utilized in the practice of cytopathology. This video has information about pap smear staining papanicolaou staining technique. Papanicolaou stain also papanicolaou s stain or pap stain is the most important stain utilized in the practice of cytopathology. Papanicolaou stain also papanicolaou s stain and pap stain is a multichromatic multicolored cytological staining technique developed by george papanicolaou in 1942. Newcomer supply papanicolaou pap stain provides three classic staining solutions for cytology preparations, allowing for crisp and distinct nuclear detail and differentially stained cytoplasm. Original article comparison of the routine papanicolaou. The slides were prepared by the clinicians immediately after fna procedure using crash and pull technique.

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