Wrights stain is named for james homer wright, who devised the stain in 1902 based on a modification of romanowsky stain. A protocol for papanicolaou staining of cytologic specimens. The bv smear will be heat fixed at the receiving laboratory before being gram stained. Acidfast stain principle, procedure, interpretation and. Leishman stain is a mixture of methylene blue, and eosin dye, prepared in alcohol medium and diluted with buffer or distilled water during staining procedure. The papanicolaou stain which aids in preserving the smears, and cells are easily described due to their consistent staining pattern. 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. Bacterial capsules are nonionic, so neither acidic nor basic stains will adhere to their surfaces. Making and staining a blood smear university of vermont. Xylene blot excess xylene before going into ethanol. First, crystal violet, a primary stain, is applied to a heatfixed smear. Fix the airdried smear specimen in methanol for 10 20 minutes 2. The slides must not be in contact with each other or else the stain will be flow off.
Hold the tube at an angle and introduce the capillary tube. A blood film or peripheral blood smear is a thin layer of blood smeared on a microscope slide and then stained in such a way to allow the various blood cells to be examined microscopically. A comparative study on staining techniques for vaginal. Ppap smearap smear vvandana mehta, vani vasanth, c. Staining procedure for thin blood film bp healthcare. After drying, the smear can be placed directly into 7qq alcohol to coagulate the albumin, prior to staining. Flood airdried, heatfixed smear of cells for 1 minute with crystal violet staining reagent. Guidelines for blood smear preparation and staining procedure.
A nuclear stain, haematoxylin, is used to stain cell nuclei. Queensland cervical screening program manual for authorised. Any modifications to the staining procedure working solutions may affect the staining result, and are subject to precise method validation. Aug 23, 2018 a pap smear test is very important to screen for abnormalities of cells in the cervix. Start studying preparation of smears and simple staining. Nov 21, 2017 if your pap smear test results are abnormal, your doctor will perform colposcopy, a procedure in which a special magnifying instrument called the colposcope is used to examine the tissues of the cervix, vulva and the vagina.
Cell staining in microscopy types, techniques, preparations and procedures. Prepare bacterial smears for the microscopic visualization of bacteria. New and reproducible modified papanicolaou staining procedure. Preparation of peripheral blood smear and staining procedure. How to stain peripheral blood and interpret blood film. Place slides containing paraffin sections in a slide holder glass or metal deparaffinize and rehydrate sections. Most information and illustrations in this book are extracted from the authors.
Flagella stain principle, procedure and result interpretation. This technique was developed by george papanicolaou, the father of cytopathology. The extra time and care taken during the field season will be rewarded later when the smears must be scanned, and parasites identified and counted. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas. The doctor may choose to do a procedure known as a colposcopy. Preparation and examination of skin smears official web.
It is a polychromatic stain containing multiple dyes to differentially stain various components of the cells. A protocol has been developed for restaining cytologic specimens that have been analyzed on a multidimensional slitscan flow system. Pap staining is used to differentiate cells in smear preparations of various bodily secretions. Smeared material should be fixed by placing the slides on an electric hotplate prior to staining 6575c. It involves examining cells taken from the cervix under a microscope. However, two modifications have been made since the introduction of bts and changes in the terminologies and definitions may cause confusion. This monograph essentials of pap smear and breast cytology is prepared at the request of a large number of students in cytology who wish to have a small and concise book with numerous illustrations for easy reference during their laboratory training. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic multicolored cytological staining technique developed by george papanicolaou in 1942. The most common cause of unsatisfactory smear on pap spin was scant cellularity in 3 cases 1. In this lesson, readers will learn about the procedure, staining methods, and interpretation of the blood smear test. The pap test is a highly complex laboratory test requiring careful patient preparation, skill in clinical collection, highly skilled and. Compare the shapes and arrangements of bacterial cells. The nucleus, which is acidic in character, has an affinity for basic stains and the.
In your procedure however, the preparation of the smear needs not be sterile but aseptically done to prevent unwanted materialsorganisms on your slide and assuring that only the specimen you intend to examine is on your slide. Your explanation of the principle and procedure for acid fast staining were basically correct. The video shows the procedure for preparation of a peripheral blood smear and its staining prior to observation under a microscope. Giemsa stain recommended for detection and identification of blood parasites. When these abnormal cells are present in the cervix, it is called cervical cancer, or cancer of the cervix. Request pdf guidelines for blood smear preparation and staining procedure for setting up an external quality assessment scheme for blood smear interpretation. Pap smear test procedure, results interpretation health. Wait until the thin and thick films are completely dry before staining. The smear and simple stains san diego miramar college. Prepare a thin smear of the material to be stained such as feces, urine, sputum, saliva. Immunohistochemical staining with cd68 was performed in. To describe the procedure for properly staining malaria blood films with giemsa stain. Immunohistochemical staining with cd68 was performed in 15 selected.
Conventional pap smear and liquid based cytology for cervical. National public health laboratory world health organization. The ideal bv smear should upon visual inspection cover at least twothirds of the surface area of a properly labeled glass slide and airdried. Making and staining a blood smear a wellmade blood smear is a beauty to behold, and likely to yield interesting and significant information for a research project. The needle is briskly redirected within the mass at several different angles.
Neel, dvm, dacvp clinical associate professor, clinical pathology nc state college of veterinary medicine raleigh, nc, 27607 introduction although tremendous advances have been made in the field of pointofcare hematology analyzers, examination of a well prepared, well stained blood smear remains the cornerstone of veterinary. Stain in hematoxylin stain, gill i or gill ii for 1 to 6 minutes, depending on preference of nuclear stain intensity. In capsule staining procedure we do not heat fix and rinse the smear with water as heat and water may dislodge capsules from bacteria. The classic form of pap stain involves five dyes in three solutions. A blood filmor peripheral blood smearis a thin layer of blood smeared on a glass microscope slide and then stained in. If a womans pap smear result is normal, she will continue routine screening. Bacterial smears and simple stains by myriam feldman issuu. Revised february 2016 papanicolaou stain for cytological. The use of tap water is not recommended when making the smears or when performing rinse steps in some staining protocols, for example, in the ziehlneelsen protocol. Check out the leishman staining technique used in hematology laboratory for the examination of peripheral blood smear. When getting it done, you will be asked to lie on an examination table during the procedure with your feet in stirrups for support and your legs spread. Preparation of blood smear with different staining method dr. Flood smear with carbol fuchsin stain zcarbol fuchsin is a lipid soluble, phenolic compound, which is able to penetrate the cell wall 3. Cover a heat fixed smear with a basic dye crystal violet.
The objective of this study was to develop a papanicolaou stain as fast as diffquik yet with cytomorphology as exquisite as that processed by thinprep for the optimal evaluation of fine needle. The definitive diagnosis of malaria infection and other parasites is still based on finding such organisms in blood films. The excess stain is removed with dilute hydrochloric acid solution acid water. One covers the fixed smear with stain for specific period, after which this solution is washed off with water and slide blotted dry. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou, the father of cytopathology. When the blood smear is totally dry, arrange the slides on the slide rack, with the blood smear facing front. Its primary purpose is to identify patients who have cellular changes that place them at risk for the development of cervical cancer. To use and relate the gram stain to the study of bacterial cell morphology, and as an important step in the identification of a bacterial species. As the drawing shows, the cervix is the lower, narrower part of the uterus. Cervical cancer basic facts on screening and the pap test.
This method enhances the nuclear component, increases cytoplasmic transparency, and reduces background staining. To learn the techniques of gram staining, nigrosin staining and koh test. Objectives to learn the technique of smear preparation. If, the smear is unfixed, fix it by gently heating the smear on flame. This technique also utilises a fine gauge needle 22 25 g. Several smear making methods and variations have been documented personal preference and teaching exposure influence the method of choice but the physical properties of the fna material including the concentration of hypocellular material or cyst fluid can also dictate the most useful method of triaging and smear making. Processing cytology samples in the laboratory eurocytology. This is an alternative stain to the chromotrope procedure that is a fast, reliable, and simple method of staining smears to demonstrate microsporidian spores in fecal and other clinical specimens. If the pap smear result is abnormal, the doctor will recommend repeat testing or more frequent followup, depending upon the exact type of abnormality and whether any infection is present. A pap smear, also called a pap test, is a routine screening procedure for cervical cancer. Women age 30 and older who have had 3 normal pap smears in a row should have a pap smear every 3 years. Put your index finger over the top of the capillary tube before removing it or blood will leak run back down into the sample. The current reporting system is the bethesda system, which was introduced in 19886 and later updated again in 1999 table 1. However, note that mast cell and basophil granules may not stain.
The test doesnt diagnose cancer, but rather looks for abnormal cervical changes cervical dysplasiaprecancerous or cancerous cells that could indicate cancer. Introduction to peripheral blood smear examination clinical. I recommend the standard 666 dipping protocol for blood smears. As such, it is used for cervical cancer screening in gynecology. Cancer is a disease in which cells in the body grow out of control. Blood smear is prepared, stained with leishmans stain and cells are identified under oil immersion lens. Notice the angle and direction that the spreader slide is pulled, first back towards the drop of blood, then quickly forward to the end of the slide. As earlier mentioned,the examination of a stained peripheral smear is an integral part of laboratory evaluation of patients and this provides information on red cells, leukocytes and platelets and is used to supplement the information provided by. The technique uses a high number of dyes in its procedure. Jan 12, 2020 procedure of regressive papanicolaou staining method. Blood smear examination is the preferred diagnostic method for certain parasitic infections, such as malaria and babesiosis.
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. Fix the thin film with methanol 100% or absolute and let it dry completely before staining. Ppap smear reportingap smear reporting the pap smear reporting classification has evolved and been refined over time. Cover each slide completely with 12ml of leishmans stain undiluted using. Blood smear 1 2 3 4a 4b 5 gently invert the tube to mix the sample.
Jul 05, 2011 instructions on preparing a bacterial smear and staining using a simple stain. 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. A6600 description fields stain is a histological method for staining of blood smears. The following points highlight the top five types of staining. If you continue browsing the site, you agree to the use of cookies on this website. Done regularly, it can help to recognize changes earlier and allows many women to seek treatment and avoid. The document covers the methods for the staining procedures commonly used in clinical microbiology laboratories for the identification of pathogens and dyes stains. Nov 03, 2016 preparation and staining of peripheral blood smear slideshare uses cookies to improve functionality and performance, and to provide you with relevant advertising. The staining procedures require the use of a mordant so that the stain adheres in layers to the flagella, allowing visualization. The improvised pap staining protocol with minimum alcohol use is a simple, costeffective and technicianfriendly procedure that can be easily adopted in highvolume, resourcelimited laboratories. Examine the stained smear under oil immersion objective x 100 of microscope. It is also used for staining bone marrow aspirates, urine samples and to demonstrate malarial parasites in blood smears.
If necessary, the smear may be left in the fixative for 3 days, but prolonged fixation affects the staining reaction. The papanicolaou stain is one of the most widely used stains in cytology, where it is used to aid pathologists in making a diagnosis. Berkan tk, reeder je, lopez pa jr, gorman km, wheeless ll jr. Learn vocabulary, terms, and more with flashcards, games, and other study tools. Giemsa and may grunwald solutions are intended for use in staining blood films or bone marrow films. It also eliminates several steps to save time and expense. Women who have had a hysterectomy for noncancerous reasons do not. Generally, a fixed smear is supplied in examination as stress is made mostly on the staining procedure. Giemsa stain is a buffered thiazineeosinate solution designed to provide coloration of blood cells similar to the original product described by giemsa. Women from ages 21 to 30 should have a pap smear every three years instead of each year unless they have had a previous abnormal pap smear within the past year. Interpretation of pap smear reports bts has been developed to improve the communication between cytopathologists and clinicians so that the latter can be better informed on the findings on the slides. Papanicolaou stain also papanicolaous stain or pap stain is the most important stain utilized in the practice of cytopathology. The preparation of a smear is required for many laboratory procedures, including the gramstain. Processing cytology samples in the laboratory processing cytology samples involves reception of the specimen and request form, preparation of slides for microscopic examination, staining, screening and reporting the slides.
Cervical cancer basic facts on screening and the pap test what is cervical cancer. Preparation of human blood smear blood cells can be identified on the basis of the staining properties of the nucleus and cytoplasm. The decolourising process is then stopped by immersing the slide in running tap water. The purpose of making a smear is to fix the bacteria onto the. Its recommended once every three years for women starting at age 21, regardless of whether or not you. Smear making and special techniques for recovering and. This picture illustrates the proper procedure for making blood smears.
A pap test is a screening test that can also be a diagnostic test. 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. Your doctor will also take a tissue sample from any areas that appear abnormal, and send it to the laboratory for a. A staining technique using modified mayers hematoxylin and supersaturated lithium carbonate for aspiration cytology and body fluid smear is described. The most useful staining procedure in medical microbiology. A properly prepared blood smear is essential to accurate assessment of cellular morphology. Do not allow smears to dry and immediately fix in 95 percent alcohol for 5 15 minutes. Balachandran iintroductionntroduction cervical cancer is predominantly sexually transmitted and the association between certain oncogenic high risk strains of human papillomavirus hpv and cervical cancer is well documented. Contents foreword 1 introduction 2 section 1 policy, protocol and procedure manual 1. A protocol for papanicolaou staining of cytologic specimens following flow analysis. The skin smear is a valuable, costeffective tool in the routine management of the patient with hansens disease. This lesson also includes some general descriptions of conditions indicated by.
Cervical cancer is one of the major causes of mortality among women worldwide. Cytology sample collection and preparation for veterinary. When using the regressive staining method, the nucleus is deliberately overstained with a nonacidified haematoxylin. Printerfriendly preparation and examination of skin smears pdf 225 kb. Using a sterilized inoculating loop, transfer loopful of liquid suspension containing bacteria to a slide clean grease free microscopic slide or transfer an isolated colony from a culture plate to a slide with a water drop. The steps of the gram stain procedure are listed below and illustrated in table 1. Pap test or pap smear procedure eperiod calculator. Wrights stain is a type of romanowsky stain, which is commonly used in hematology laboratory for the routine staining of peripheral blood smears.
Staining methods histology and cytology 152 notes sources of errors irregular distribution of the blood smear on a glass slide may result in an erroneous cell counts. Alcohols used for wiping the skin may cause hemolysis and artifacts. Distinguishes bacteria of two large and medically important groups. Bacterial smears and simple stains escherichia coli. Introduction smears much of your success in staining microbes will come from preparing a good smear. The document also includes the 9 standard operating procedures for the pap staining in cervical. Here, the methods for making and staining smears are given, as well as a list of sources for high quality slides, stain, and chemicals. Pap smear staining papanicolaou staining technique.
Do not let the specimens dry at any stage of the staining procedure. Please note that the quality of the smear too heavy or too light cell concentration will affect the gram stain results. Gyenocologic pap test collection procedure medical center. The fine needle aspiration procedure is less likely to result in adverse effects when compared to tissue biopsy. It should be done by careful look on the narrow edge of the slide or by moving a thumb smoothly on its edge. The procedure of flagella stain wet mount technique grow the organism to be stained at room temperature on blood agar for 16 to 24 hours. The first page of the pdf of this article appears above. Here a syringe is not attached to the needle for aspiration of skin masses.
316 344 1284 908 1428 1179 1361 877 893 1319 640 1012 1204 436 43 717 678 698 1270 394 1534 941 567 541 781 789 56 491 246 792