It is also used for staining bone marrow aspirates, urine samples and to demonstrate malarial parasites in blood smears. If you continue browsing the site, you agree to the use of cookies on this website. A fresh, wellmade, peripheral blood film is crucial for accurate cell morphology assessment. Many hematological diagnoses depend upon this procedure and often a definitive diagnosis can be established from the careful examination of the blood film. 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. Giemsa stain protocol for visualization of blood microorganisms. Peripheral blood smear procedure and interpretation. Blood smears are needed for microscopic examination of the blood. An atlas of blood cells may be given out in class you may keep it or will be found in your textbook 3. A simple single differential stain of blood smears using. Prepare a film of blood or bone marrow on a microscopic slide and allow to air dry. Wewilluseadropletofbloodtomakeathin%smear,dryit,fixandstainitand observe under a microscope. Validation and optimization of criteria for manual smear.
Hematology sequence blood labs orientation objectives objectives procedure 1. It is hoped that this paper will provide an aid to those who wish to employ the blood smear as a diagnostic tool. In nutshell, the value of peripheral blood smear in assessing morphology and differential counting of blood cellular elements cannot be downplayed. 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.
A phlebotomist, a person specifically trained to draw blood, first cleans and sterilizes the injection site with an antiseptic. A peripheral blood smear examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide. Stained peripheral blood smear is examined carefully using 40 x to 100 x objective usually oil immersion lens 2. Wait until the thin and thick films are completely dry before staining. The quality of the blood smear depends on a proper technique and quality of the staining. The mechanism of action of wrights stain is also discussed. The drop of blood used for a blood smear test contains numerous red blood cells rbcs, white blood cells wbcs, and platelets. It is good practice to prepare two slides for each sample. 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 advantages of using this combination are that the procedure can be standardized and produces consistent results from batch to batch.
The test provides information on the number and shape of these. A blood smear is a blood test used to look for abnormalities in blood cells. This picture illustrates the proper procedure for making blood smears. Microscopy of peripheral blood thin and thick films remains the reference for malaria diagnosis. 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. A properly prepared blood smear is essential to accurate assessment of cellular morphology. 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. The smear is greater than 25 mm long and the feathered edge stops approximately 10 mm from the end of the slide. The slide is the stained and viewed under a microscope. This procedure is intended for use in malaria clinical research studies such as those assessing drug or vaccine ef cacy. Introduction to peripheral blood smear examination. The slides should be dried as quickly as possible to prevent.
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. Wrights stain is a type of romanowsky stain, which is commonly used in hematology laboratory for the routine staining of peripheral blood smears. For these aspects, the authors recommend reference to existing islhicsh guidelines and standard morphology textbooks 1 5. Some additional guidelines should be followed to create the. Although there are several techniques described for making blood smears, most. Jan 17, 2017 the video shows the procedure for preparation of a peripheral blood smear and its staining prior to observation under a microscope. Delay in preparation of smears can result in changes in parasite morphology and staining characteristics. Place the airdried smear on the slide staining rack, smear side facing upwards. Making and staining a blood smear university of vermont. 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.
Select two glass slides that are clean and free of chipped edges. Blood smears, at least two thick and two thin, should be prepared as soon as possible after collection. Making a quality blood smear although there are several techniques described for making blood smears, most people use the wedge or push technique. Check out the leishman staining technique used in hematology laboratory for the examination of peripheral blood smear.
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. This recommended protocol is applicable for the visualization of blood parasites. The examination of blood films stained with wrights stain frequently provides important clues in the diagnosis of anemias and various. Video is prepared by the faculty and students of the department. Blood smear examination is the preferred diagnostic method for certain. In the present study, out of total 218 cases tested, 19. The interference of moisture in leishman stain is a wellknown problem in tropical climate and in rainy season, even in most wellequipped laboratories. The examination of a peripheral blood smear is one of the most informative exercises a physician can perform. 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. Always start with room temperature, wellmixed, clot free, edta anticoagulated blood heparin is used for some exotic species.
It is not appropriate to use the same edge of a slide for more than a single 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. Pap staining is used to differentiate cells in smear preparations of various bodily secretions. 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. 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.
Learn preparing staining a blood smear with free interactive flashcards. Megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation. 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. The examination of the peripheral blood smear is an important basic hematological procedure. Extremely thin films caused by too small a drop, too slow. In this lesson, readers will learn about the procedure, staining methods, and interpretation of the blood smear test. Cover the blood film with undiluted staining solution. The undiluted stain fixes and partially stains the smear. A blood filmor peripheral blood smearis a thin layer of blood smeared on a glass. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas.
This lesson also includes some general descriptions of conditions indicated by. Cytology sample collection and preparation for veterinary. Seed haematology the role of the peripheral blood smear. The differential staining allows one to identify the types of white blood cells on the smear. If the blood has been refrigerated, allow it to return. After one minute staining with black quink, the walls of the red blood cells erythrocytes. It is then treated with a special stain and studied under a microscope to examine the blood cells. An expert examination of the blood smear can identify errors, establish a diagnosis, or lead to a useful fortuitous finding. Plasmodium vivax centers for disease control and prevention. 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. The peripheral blood film remains a very useful tool in hematological assessment. 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.
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. Peripheral blood smear test purpose, procedure, results. Second, appearance of the organism in peripheral blood, especially in cats, can be. Introduction to peripheral blood smear examination clinical. How to stain peripheral blood and interpret blood film.
If there will be a delay in staining smears, dip the thick smear briefly in water to. Blood smears are used to determine leukocyte differentials, to evaluate erythrocyte, platelet and leukocyte morphology, and, if necessary, to estimate platelet and leukocyte counts. Making the blood smear before any stained smear can be used for a diagnosis. Choose from 500 different sets of preparing staining a blood smear flashcards on quizlet. It works by precipitating proteins and carbohydrates. Preparation of peripheral blood smear and staining procedure.
Pdf analyzing microscopic images of peripheral blood. Papanicolaou stain also papanicolaous stain and pap stain is a multichromatic staining histological technique developed by george papanikolaou, the father of cytopathology. Notice the angle and direction that the spreader slide is pulled, first back towards. Using the corner of a clean slide, spread the drop of blood in a circle the size of a dime diameter 12 cm. A peripheral blood smear or peripheral blood film is a glass slide covered on one side with a thin layer of venous blood. 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. The giemsa blood smear test is a test to detect blood parasites. Staining of pbf and notes interpretation of normal. Review of platelet estimates medical laboratory observer. 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. The three main blood cells that the test focuses on are. 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 drop should be in the center line of the glass slide. Preparation of a peripheral blood film this protocol describes how to prepare a slide film of peripheral blood for microscopic examination.
Jul 15, 2019 a peripheral blood smear examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide. 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. Leishman stain is the universal stain used in staining of peripheral blood smears all over the world. Prepare a second smear by repeating steps 35, using the edge of the first slide to spread a drop of blood on the second slide. The video shows the procedure for preparation of a peripheral blood smear and its staining prior to observation under a microscope. Cell fixationis done by placing the slide in methanol ch3oh. A well prepared blood smear is necessary for microscopic examination of blood. An atlas of instructive blood smears is included as a set of slides. Do not make the smear too thick or it will fall off the slide. 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.
Seed haematology the role of the peripheral blood smear in. 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. Staining of pbf and notes interpretation of normal and. The art of peripheral blood smear preparation, staining, and examination is losing its shine. Most commonly blood smears are used for the differential leukocyte count dlc. Microscopic exam of a peripheral blood smear several parts 2. Peripheral blood smear can be used for est imation of. Here, the methods for making and staining smears are given, as well as a list of sources for high quality slides, stain, and chemicals. Preparation of blood smear with different staining method dr.
Blood films that are too thin or too thick present a problem. A blood film or peripheral blood smear is a thin layer of blood smeared on a microscope slide. With the advent of hematological automated machines, complete and highly accurate hematological data and parameters can be obtained within a few minutes. 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. Blood filled capillary tube or purple top with diffsafe dispenser. Although advances in automated technology have made the examination of a peripheral blood smear by a physician seem less important, the techno. Right blood as seen at 400x under light microscope. Peripheral blood or potassium edta anticoagulated blood 12 mg edta1 ml blood may be used. 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. The examination of a stained peripheral blood film is a hematology test and is an integral part of laboratory evaluation of patients.
Wrights stain is named for james homer wright, who devised the stain in 1902 based on a modification of romanowsky stain. In such a scenario, troubleshooting for suboptimal staining becomes challenging. 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. Capillary blood should be obtained by fingerstick, or venous blood should be obtained by venipuncture.
If you are using venous blood, blood smears should be prepared as soon as. The smear is stained with wrights stain and performed after the complete blood count is run. Blood from a patient with anaemia will have a lower. Smears made from edtaanticoagulated blood should be made. Giemsa stain is a classical blood film stain for peripheral blood smear and bone marrow. The peripheral blood film pbf is a laboratory workup that involves cytology of. 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.
Although giemsa staining is most commonly used, the leishman staining method provides better visualization of the nuclear chromatin pattern of cells. To study the effect of incubation of buffer solution, slide alone and slide. 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. Thick smears consist of a thick layer of dehemoglobinized lysed red blood cells rbcs. See the article on artifacts in peripheral blood elsewhere in this issue for a. The examination of a stained peripheral blood film is a hematology test and is. The purpose of preparation of blood smear blood film is to study the morphology of rbcs, differential leukocyte count and reticulocyte count.
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