It can also reveal changes in your cervical cells that may turn into cancer later. In the united states there are approximately 20,000 new cases of cervical cancer. The health care professional first places a speculum inside the vagina. 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. The papanicolaou pap stain kit cytology stain is designed to differentiate between a variety of cells in vaginal smears for detection of vaginal, uterine and cervical cancer. The papanicolaou staining procedure is used for examining exfoliated cells in secretions, exudates, transudates or scrapings of various internal organs and tissue. Pdf modified ultrafast papanicolaou staining technique. Onethousand and sixty women were enrolled in this study. Jan 12, 2020 papanicolaou stain also papanicolaous stain or pap stain is the most important stain utilized in the practice of cytopathology.
The purpose of this study was to examine the characteristics of gram stain versus pap smear in diagnosis of bacterial vaginosis bv. An intro to routine and special staining in histopathology. In addition, this procedure is valuable for staining a variety of other bodily secretions and cell smears. Pap smear screening has been documented to decrease the incidence and mortality rate of cervical cancer. Gail curtis, in essential clinical procedures second edition, 2007. Nov 05, 2018 pap smear staining, pap smear staining procedure, pap smear procedure steps, pap smear staining method, pap smear staining procedure ppt, pap smear staining procedure pdf, how to do pap smear. Submerge successively in alcohol 80%, alcohol 70%, alcohol 50% and water, 1 minute in each liquid. Place the slide in the stain, thick film down to prevent the debris caused by dehemoglobinization from falling onto the thin film. Romanowsky stains may also be used for wet fixed slides, but are primarily applied to airdried smears. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic staining. Hematoxylin, a common nuclear stain, is isolated from an extract of logwood haematoxylon campechianum1 the first successful bi.
It is employed in the pap stain to demonstrate keratin. This item has been discontinued and is no longer available. A pap smear is a simple, quick, and relatively painless. Procedure cytoplasmic stain nuclear stain optimal suboptimal optimal suboptimal papanicolaou no of smears 110 90 20 100 10 reap no. Change alcohols and masterclear when discolored for best stain clarity. Two changes of 95%ethyl alcohol with standard pap stain after og6 are replaced by 1% acetic acid 10dips. Stain in papanicolaou stain, og6 for 30 seconds to 2 minutes, depending on specimen type and preference of stain intensity. Papanicolaou stain includes both acidic and basic dyes. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic multicolored cytological staining technique developed by george papanicolaou in 1942. Understaining is common when a leukemia slide is placed on an automated slide stainer. Papanicolaou stain an overview sciencedirect topics. Choice of fixative and technique is dependent on specimen types analyzed in the laboratory. It has affinity for chromatin, attaching to sulphate groups on the d. After haematoxylin staining in the pap stain procedure, og6 is the first counterstain to be used and must be performed before the ea50 or ea65 counterstain as these will prevent satisfactory take up of og6 if they are preformed ahead of it.
Blue slides in lithium carbonate, saturated aqueous 12215 or. Papanicolaous stain pap stain is widely used in all cervical smear and nongynaecologic exfoliative smears. Stain with hematoxylin harris solution for approximately 5 minutes. Guidelines of the papanicolaou society of cytopathology for. Fill out the specimen manifest, including all areas that are not shaded. Natural dye hematoxylin is the nuclear stain which stains cell nuclei blue. Consequently, to address the shortcomings of the pap test, a number of derivative methods were developed, such as the ultrafast staining technique, the shortduration papanicolaou stain, the reap.
Siobhan bourke discusses and demonstrates correct technique for taking cervical samples. Papanicolaou stain pap stain protocol ihc world, llc. Pap smear as the cellular structure is better preserved because the cells are fixed immediately. Papanicolaou stain for cytological preparations newcomer supply. The papanicolaou stain is one of the most widely used stains in cytology, where it is used to aid pathologists in making a diagnosis. A pap test, also called a pap smear, is an exam a doctor uses to test for cervical cancer in women. The practice of retaining the filter in the filtration apparatus throughout the staining procedure minimizes cell loss and eliminates specimen cross contamination. Mdss gmbh schiffgraben 41 30175 hannover, germany symbols. The main disadvantages of the manual technique are the increased risk of spilling the stain and the longer time required to complete the procedure. Reap technique is the recent modification of standard pap technique. Remove all lids and place each in its slot to the right of the jar. Papanicolaou stain also papanicolaous stain or pap stain is the most important stain utilized in the practice of cytopathology.
Gill hematoxylin solutions are nuclear stains intended for use in histology and cytology. This is a transparent stain and gives very good cellular differentiation and nuclear details. The speculum is a metal or plastic instrument that keeps the vagina open so that the cervix can be seen clearly. The principle of pap stain is to clearly distinguish between basophilic and acidophilic cell components and obtain a detailed chromatin pattern. Papanicolaou, papanicolaou stain underwent various modifications. Revised february 2016 papanicolaou stain for cytological. All cases with symptoms of bv were determined by amsels criteria, which were accepted as the gold standard for diagnosis of bv. This technique is best suited for lowvolume laboratories. The 5dye pap stain method evolved through 2 salient phases. Process slides though solutions following the directions indicated by the arrows. Papanicolaou stain, ea, is comprised of a combination of two dyes. May 10, 2020 wikihow is a wiki, similar to wikipedia, which means that many of our articles are cowritten by multiple authors. Alum acts as mordant and hematoxylin containing alum stains the nucleus light blue. Stain the entire slide with diluted giemsa stain 1.
When a stain is complete the section is covered with a coverglass that makes the preparation permanent. The procedure of giemsa staining varies as per the purpose of staining that means whether the staining is done for the examination of blood cells or to find the parasites in the blood smear and accordingly the blood smears are prepared as thin blood films or thick blood films. Pap smear staining papanicolaou staining technique. A recommended staining protocol can be found below. In standard pap stain final dehydration is done by two changes of absolute alcohol. Papanicolaou tests also known as pap tests are named after their inventor, georgios papanicolaou, and serve to detect hpvassociated lesions in the cervix. As such, it is used for cervical cancer screening in gynecology. For tap water rinse steps, submerse slide into dish filled with tap water and then place the dish under a gently running tap for the time indicated. The pelvic examination and obtaining a routine papanicolaou smear. Stain in papanicolaou stain, ea36, ea50 or ea65 of choice for minutes, depending on specimen type and preference of stain intensity.
Original article comparison of the routine papanicolaou. This technique was developed by george papanicolaou, the father of cytopathology. Harris hematoxylin is the optimum nuclear stain zmaygrunwald giemsa stain is one of the common romanwsky stains used in cytology. Oct 30, 2019 the pap test is a procedure that collects cells from the cervix so that they can be looked at closely in the lab to find cancer and precancer. The technique uses a high number of dyes in its procedure.
Cells are fixed to a slide and stained first with hematoxylin, which stains the nuclei followed by og6 and ea50 or ea65 as a counterstain. Place slides in the jar of 95% alcohol next to the start arrow lower left corner of tray. It is useful for studying cell morphology in airdried smears. The procedure was developed in the early 1940s by george. This chapter presents the principle and steps of the routine stains in cytology laboratory. This is always done as part of the staining process. Comparison of gram stain and pap smear procedures in the. The polychromatic pap stain involves five dyes in three solutions. Label the requisition and specimen containersslidewith the patients name and a second identifier e. Inception and development of the papanicolaou stain method. Residents ppap smearap smear bioline international.
Blood, bone marrow, or tissue touch preparations standard procedure. This uses a combination of two dyes, hematoxylin and eosin used for demonstration of nucleus and cytoplasmic inclusions in clinical specimens. Papanicolaou pap red stain kit cytology stain abcam. Uf pap, rapid pap, and rapid economic acetic acid pap stain have been introduced in 1995, 2004, and 2008, respectively. Cyto prep techniques questions and study guide quizlet. Routine staining in cytology laboratory springerlink. Development of the pap stain method followed specific historical and scientific events. Singh et al pap staining for sperm morphologic analysis features in smears.
The first staining step is dewaxing which uses a solvent to remove the wax from the slide prior to staining. Papanicolaou stain a papanicolaou stain is typically performed in the laboratory on alcoholfixed slides. The papanicolaou stain is recommended for the staining of alcohol fixed cytology slides. Papanicolaou staining is the universal method for staining gynecologic slides. The classic form of pap stain involves five dyes in three solutions. Thermo scientific rapidchrome papanicolaou staining kit. Chlamydia testing using floq and uriswabs is also discussed. Shake slide gently between changes to remove excess liquid and prevent carry over into next dish. Papanicolaou staining is applied to vaginal exudates for the detection of uterine or. Learn vocabulary, terms, and more with flashcards, games, and other study tools.
It is a polychromatic stain containing multiple dyes to differentially stain various components of the cells. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou, the father of cytopathology. Prostatic acid phosphatase pap, also prostatic specific acid phosphatase psap, is enzyme produced by prostate tissue acid phosphatases release phosphate groups, optimally at acid ph may promote hiv infection. To create this article, volunteer authors worked to edit and improve it over time. Cytologists rely heavily on the quality and appearance of the stain.
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. In conventional cytology, preparation of a specimen by wet fixation for papanicolaou stain is potentially subject to dry effect or cell loss which may make cytologic interpretation. This stain reveals nuclear detailsmembrane irregularities, chromatin pattern, and intranuclear invaginationsimportant for establishing a diagnosis of malignancy, especially welldifferentiated neoplasms that lack significant pleomorphism. Evolution of pap stain biomedical research and therapy. Ea36, ea50 and ea65 denote the varying proportions of dyes in each solution and the formula of choice will depend upon a laboratories staining preference. Comparison of ultrafast papanicolaou stain with the standard. Importance of papanicolaou staining for sperm morphologic. The bd surepath liquidbased pap test helps detect disease with a more quality and easily read sample than possible with conventional pap smears and other liquidbased cytology lbc tests.
A nuclear stain, haematoxylin, is used to stain cell nuclei. The pap stain, ea also varies with each ea formulation made with either light green sf or modified with fast. Pap staining is used to differentiate cells in smear preparations of various bodily secretions. The pas staining procedure may also be used for the demonstration of fungal organisms in tissue sections. On implementation, this relatively inexpensive cytology test dramatically reduced the incidence and mortality rates of cervical cancer. Stain in hematoxylin stain, gill i or gill ii for 1 to 6 minutes, depending on preference of nuclear stain intensity.
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