Making and staining a bloo d smear a wellmade blood smear is a beauty to behold, and likely to yield interesting and significant information for a research project. The procedure for performing a platelet estimate varies, depending on how the smear is prepared wedge method, semiautomated, or automated, the type of specimen collected edtaanticoagulated blood vs. Making and staining a blood smear university of vermont. 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. A phlebotomist, a person specifically trained to draw blood, first cleans and sterilizes the injection site with an antiseptic. An expert examination of the blood smear can identify errors, establish a diagnosis, or lead to a useful fortuitous finding. It works by precipitating proteins and carbohydrates.
Blood smears are needed for microscopic examination of the blood. Jul 15, 2019 a peripheral blood smear examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide. The blood samples collected from 218 clinically suspected cases of malaria were subjected to giemsa staining, acridine orange staining and rapid diagnostic test rdt as per standard procedures. The purpose of preparation of blood smear blood film is to study the morphology of rbcs, differential leukocyte count and reticulocyte count. It is also used for staining bone marrow aspirates, urine samples and to demonstrate malarial parasites in blood smears. The examination of a peripheral blood smear is one of the most informative exercises a physician can perform. The giemsa blood smear test is a test to detect blood parasites. In this lesson, readers will learn about the procedure, staining methods, and interpretation of the blood smear test. A blood film or peripheral blood smear is a thin layer of blood smeared on a microscope slide. The test provides information on the number and shape of these.
The smear is greater than 25 mm long and the feathered edge stops approximately 10 mm from the end of the slide. Blood films that are too thin or too thick present a problem. Although there are several techniques described for making blood smears, most. See the article on artifacts in peripheral blood elsewhere in this issue for a. Select two glass slides that are clean and free of chipped edges.
Staining of pbf and notes interpretation of normal. A fresh, wellmade, peripheral blood film is crucial for accurate cell morphology assessment. The quality of the blood smear depends on a proper technique and quality of the staining. Preparation of blood smear with different staining method dr. Cell fixationis done by placing the slide in methanol ch3oh. 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.
Place the airdried smear on the slide staining rack, smear side facing upwards. Stained peripheral blood smear is examined carefully using 40 x to 100 x objective usually oil immersion lens 2. 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. Preparation of a peripheral blood film this protocol describes how to prepare a slide film of peripheral blood for microscopic examination. After one minute staining with black quink, the walls of the red blood cells erythrocytes. The procedure of leishman staining may vary as per the purpose of staining that means whether the staining is done for the examination of blood cells morphology, toxic granules in leucocytes, type of anemia etc. How to stain peripheral blood and interpret blood film.
Wewilluseadropletofbloodtomakeathin%smear,dryit,fixandstainitand observe under a microscope. This picture illustrates the proper procedure for making blood smears. A peripheral blood smear pbs or blood film is required to be made from capillary blood or from a drop of blood from an edta anticoagulated blood sample. 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. It is then treated with a special stain and studied under a microscope to examine the blood cells.
The undiluted stain fixes and partially stains the smear. Thick smears consist of a thick layer of dehemoglobinized lysed red blood cells rbcs. The slide is the stained and viewed under a microscope. Review of platelet estimates medical laboratory observer.
The slides should be dried as quickly as possible to prevent. Making a quality blood smear although there are several techniques described for making blood smears, most people use the wedge or push technique. An atlas of blood cells may be given out in class you may keep it or will be found in your textbook 3. Giemsa stain is a classical blood film stain for peripheral blood smear and bone marrow. Blood smear examination is the preferred diagnostic method for certain. 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. This way, each slide may be used to spread the film of the other, and duplicate slides will be of value if something happens to one of them in the. Jan 17, 2017 the video shows the procedure for preparation of a peripheral blood smear and its staining prior to observation under a microscope. A blood filmor peripheral blood smearis a thin layer of blood smeared on a glass microscope slide and then stained in such a way as to allow the various blood cells to be examined microscopically. Cover the blood film with undiluted staining solution. After staining the smear with fields stain,the slide is observed under microscope for staining characteristic using 100x objective with cedar wood oil or immersion oil. Peripheral blood smear test purpose, procedure, results.
Always start with room temperature, wellmixed, clot free, edta anticoagulated blood heparin is used for some exotic species. It is hoped that this paper will provide an aid to those who wish to employ the blood smear as a diagnostic tool. Most commonly blood smears are used for the differential leukocyte count dlc. An unstained blood smear is clearly seen by eye on the microscope slide, however, the blood cells are barely visible under the microscope figure 1. The interference of moisture in leishman stain is a wellknown problem in tropical climate and in rainy season, even in most wellequipped laboratories. It is used to diagnose diseases, such as malaria, which are caused by parasitic infections the collected blood sample is smeared onto a glass slide and stained with giemsa stain. Hematology sequence blood labs orientation objectives objectives procedure 1.
Here, the methods for making and staining smears are given, as well as a list of sources for high quality slides, stain, and chemicals. This section of the document describes the procedure for the preparation of blood lms from peripheral blood collected by ngerprick or venepuncture for staining and microscopy to detect, identify and quantify malaria parasites. Right blood as seen at 400x under light microscope. Do not make the smear too thick or it will fall off the slide. In such a scenario, troubleshooting for suboptimal staining becomes challenging. It is not appropriate to use the same edge of a slide for more than a single film. Extremely thin films caused by too small a drop, too slow. Blood smears are used to determine leukocyte differentials, to evaluate erythrocyte, platelet and leukocyte morphology, and, if necessary, to estimate platelet and leukocyte counts. Attention to detail in slide preparation, staining and standard criteria for cell estimates, morphology and differential reporting enhance the value of results provided to the physician. In the present study, out of total 218 cases tested, 19. The smear is stained with wrights stain and performed after the complete blood count is run. An atlas of instructive blood smears is included as a set of slides. A peripheral blood smear or peripheral blood film is a glass slide covered on one side with a thin layer of venous blood.
Learn preparing staining a blood smear with free interactive flashcards. Wrights stain is a type of romanowsky stain, which is commonly used in hematology laboratory for the routine staining of peripheral blood smears. Microscopic exam of a peripheral blood smear several parts 2. Plasmodium vivax centers for disease control and prevention. The mechanism of action of wrights stain is also discussed. This procedure is intended for use in malaria clinical research studies such as those assessing drug or vaccine ef cacy. 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. Many hematological diagnoses depend upon this procedure and often a definitive diagnosis can be established from the careful examination of the blood film. Blood smear technique aspirated material may contain enough blood andor liquid to allow smearing of the material in a similar fashion to that used to make a peripheral blood smear. A properly prepared blood smear is essential to accurate assessment of cellular morphology. If you are using venous blood, blood smears should be prepared as soon as possible after collection delay can result in changes in parasite morphology and staining characteristics. If there will be a delay in staining smears, dip the thick smear briefly in water to.
A simple single differential stain of blood smears using. The art of peripheral blood smear preparation, staining, and examination is losing its shine. Peripheral blood smear can be used for est imation of. The examination of a stained peripheral blood film is a hematology test and is. Cytology sample collection and preparation for veterinary. Wrights stain is named for james homer wright, who devised the stain in 1902 based on a modification of romanowsky stain. A well prepared blood smear is necessary for microscopic examination of blood. Megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation. Introduction to peripheral blood smear examination. Some additional guidelines should be followed to create the. To study the effect of incubation of buffer solution, slide alone and slide. The advantages of using this combination are that the procedure can be standardized and produces consistent results from batch to batch.
Seed haematology the role of the peripheral blood smear in. Introduction to peripheral blood smear examination clinical. The video shows the procedure for preparation of a peripheral blood smear and its staining prior to observation under a microscope. Validation and optimization of criteria for manual smear. Microscopy of peripheral blood thin and thick films remains the reference for malaria diagnosis. Smears made from edtaanticoagulated blood should be made. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou, the father of cytopathology. This recommended protocol is applicable for the visualization of blood parasites. If you are using venous blood, blood smears should be prepared as soon as. Wait until the thin and thick films are completely dry before staining. The three main blood cells that the test focuses on are. In nutshell, the value of peripheral blood smear in assessing morphology and differential counting of blood cellular elements cannot be downplayed.
A peripheral blood smear examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide. The blood film occupies the central portion of the slide and has definite margins on all sides that are accessible to examination by oil immersion. Delay in preparation of smears can result in changes in parasite morphology and staining characteristics. The peripheral blood film pbf is a laboratory workup that involves cytology of. Prepare a film of blood or bone marrow on a microscopic slide and allow to air dry. 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.
If you continue browsing the site, you agree to the use of cookies on this website. Video is prepared by the faculty and students of the department. Seed haematology the role of the peripheral blood smear in the modern haematology laboratory 2 sysmex educational enhancement and development february 20 the viscosity haematocrit of the blood, which can be highly variable from patient to patient, will also affect the smear. A blood filmor peripheral blood smearis a thin layer of blood smeared on a glass. Peripheral blood smear clinical methods ncbi bookshelf.
The examination of blood films stained with wrights stain frequently provides important clues in the diagnosis of anemias and various. The examination of a stained peripheral blood film is a hematology test and is an integral part of laboratory evaluation of patients. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas. Although giemsa staining is most commonly used, the leishman staining method provides better visualization of the nuclear chromatin pattern of cells. Capillary blood should be obtained by fingerstick, or venous blood should be obtained by venipuncture.
Seed haematology the role of the peripheral blood smear. Pdf analyzing microscopic images of peripheral blood. Check out the leishman staining technique used in hematology laboratory for the examination of peripheral blood smear. For these aspects, the authors recommend reference to existing islhicsh guidelines and standard morphology textbooks 1 5.
Pap staining is used to differentiate cells in smear preparations of various bodily secretions. This lesson also includes some general descriptions of conditions indicated by. Giemsa stain protocol for visualization of blood microorganisms. A blood smear is a blood test used to look for abnormalities in blood cells. Using the corner of a clean slide, spread the drop of blood in a circle the size of a dime diameter 12 cm. Choose from 500 different sets of preparing staining a blood smear flashcards on quizlet. Peripheral blood or potassium edta anticoagulated blood 12 mg edta1 ml blood may be used. If the blood has been refrigerated, allow it to return. Preparation of peripheral blood smear and staining procedure. The peripheral blood film remains a very useful tool in hematological assessment. Making the blood smear before any stained smear can be used for a diagnosis. Peripheral blood film or peripheral blood smear or peripheral smear examination forms an important and basic lab investigation for diagnosing a number of hematological blood disorders. The drop of blood used for a blood smear test contains numerous red blood cells rbcs, white blood cells wbcs, and platelets. The examination of the peripheral blood smear is an important basic hematological procedure.
It is not the purpose of this guideline to provide recommendations for standardized blood film preparation, staining and examination or an exhaustive description of blood cell morphology. 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. Blood films are examined in the investigation of hematological blood disorders and are routinely employed to look for blood parasites, such as those of malaria and filariasis. Blood smears, at least two thick and two thin, should be prepared as soon as possible after collection. The differential staining allows one to identify the types of white blood cells on the smear. Blood from a patient with anaemia will have a lower. It is good practice to prepare two slides for each sample. Peripheral blood smear procedure and interpretation. Although advances in automated technology have made the examination of a peripheral blood smear by a physician seem less important, the techno. Comparing leishman and giemsa staining for the assessment of. Staining of pbf and notes interpretation of normal and. Second, appearance of the organism in peripheral blood, especially in cats, can be. Blood filled capillary tube or purple top with diffsafe dispenser. With the advent of hematological automated machines, complete and highly accurate hematological data and parameters can be obtained within a few minutes.
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