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. Transformation of membrane nanosurface of red blood cells. Ideally, submit blood for a hemogram with 2 freshly prepared blood smears or please provide results from a recent hemogram within. Using a capillary tube, place a 23mm drop of blood about 1cm from the frosted end of a clean slide that is on a flat surface. A series of five slides was prepared from each sample of blood. It might be used to help identify andor keep an eye on numerous conditions that impact blood cell populations. This occurs when there is inadequate contact with the stain solutions. Below is a stepbystep guide to performing blood smears for diagnostic use. Preparation thin blood films and giemsa staining projectstudy.
Peripheral blood smear test purpose, procedure, results. Scope and application this procedure describes the preparation of thin blood films thin blood films are used differential leucocyte count, the detection of abnormal erythrocytes. The manual examination of these images is tedious, timeconsuming and suffers from interobserver variation. It should be done by careful look on the narrow edge of. Counting of rbcs and wbcs in noisy normal blood smear. A carefully prepared blood smear is vital for an accurate differential count. It should be done by careful look on the narrow edge of the slide or by moving a thumb smoothly on its edge. If using a needle and syringe, first remove the needle and then touch the end of the syringe to the slide.
Find a good area of the slide then scan at 25x or 50x examining cells at 100x as needed. Previously used or washed slides may be contaminated with residual detergents or microscopic debris that can appear as artifacts in the blood smear. Preparation of differential blood smears if you order a cbc with differential and the sample will not arrive at uvmmc within 4 hours, a blood smear must be made for the differential. A simplified wrights stain technique for routine avian blood smear staining1. Deriving full value from blood smear examination requires a wellprepared, wellstained blood smear and some basic skills in the methods of assessment.
Merging results from multiple slides allows differential reporting on the. The three main blood cells that the test focuses on are. Blood films are examined in the investigation of hematological blood disorders and are routinely employed to look for blood parasites, such. The most common method of blood smear preparation is the wedge technique, in which a drop of blood is placed near the end of one glass slide. The nucleus, which is acidic in character, has an affinity for basic stains and the. Blood cells are the cells which are produced during hematopoiesis and found mainly in the blood. The blood droplet on the left is too big and will result in a thick smear. Plasmodium ovale centers for disease control and prevention.
Glass slides should be free of chipped edges, glass fragments, dust, powder, fingerprints, and other debris, as these can affect blood spreading. Preparation of peripheral blood smears exercise 10. 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. Blood from a patient with anaemia will have a lower. Submit 3 or more freshly made rapidly dried unstained bone marrow smears with the sample for routine bone marrow examination. Prepare a blood smear according to the instructions. This 2nd edition of standard manual for laboratory technicians on sputum smear microscopy is one of the great efforts to improve the qa for sputum microscopy in the hospital. Do not smear or rub the swab in a back and forth motion as this will cause cell damage lysis. An illustrated guide to the morphology of blood cells, atlas of canine and feline peripheral blood smears covers patient assessment for common hematologic disorders and diseases in dogs and cats. Peripheral blood or potassium edta anticoagulated blood 12 mg edta1 ml blood may be used. 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. If preparation is not fixed or sticked over the slide, the cells are washed off during staining. Blood smear examinationmanual differential leukocyte count with complete. Learn preparing staining a blood smear with free interactive flashcards.
A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas. The preparation of standard lms is aided by the use of slide templates that indicate the placement of the lms and their size. A preparation of blood smear selection of a spreader. It is then treated with a special stain and studied under a microscope to examine the blood cells. Although microscopic examination of thick and thin blood films re mains the. Smears are prepared by gently rolling swab over glass slide. 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. Three mechanical devices for preparing blood smears were compared to the traditional coverslip and manual wedge methods. Smears made from edtaanticoagulated blood should be made. Pdf purpose and criteria for blood smear scan, blood smear.
Peripheral blood smear can be used for est imation of. Samples for flow cytometry cornell university college of. Thick blood film samples a relatively large volume of blood thus allowing more efficient detection of parasites increased sensitivity. The examination of the peripheral blood smear is an important basic hematological procedure. A single smear can be made per slide smear running the length of the slide or two or even three smears can share a slide, with the smears running the width of the slide. If the smear is made too thin, there is a chance the larger cells will be collected. Slides should not be left uncovered on top of the counter.
Dec 08, 2006 a well prepared blood smear is necessary for microscopic examination of blood. These are the same cases that you studied in the two labs the normal smear is in there as well. Jun 12, 2016 1 introduction to peripheral blood smear examination. Preparation of a peripheral blood film this protocol describes how to prepare a slide film of peripheral blood for microscopic examination. Criteria for blood smears if a delay in test performance is anticipated, two well made slides are requested for cbcs, platelet counts, differentials, and rbc morphology evaluation. Atlas of canine and feline peripheral blood smears. Jan 17, 2017 the video shows the procedure for preparation of a peripheral blood smear and its staining prior to observation under a microscope.
Most romanowsky stains are prepared in methyl alcohol so that they combine fixation and staining. The sample can be mixed by gently inverting the tube several times before slide preparation. One requires slides, pipettecapillary tube and blood spreader to make pbf smear. Proper blood smear preparation texas veterinary medical. Examination of blood smears is an integral part of haematological testing. Jul 15, 2019 a peripheral blood smear examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide. Factors that influence making a blood smear size of the drop. The direct microscopic visualization of the malarial parasite on the thick andor thin blood smears has been the gold standard for malaria diagnosis. Label precleaned slides preferably frostedend with patients name or other identifier, date and time of collection. Note that the diameter of the red blood cell is similar to the diameter of the small lymphocyte nucleus. Jan 09, 2017 a blood smear is often used as a followup test to abnormal results on a total blood count cbc to evaluate the various types of blood cells. If you dont know how to prepare it, check out the preparation of thick blood smear.
The accumulation of hemin causes destruction of red blood cells rbc membranes. This new methylene bluestained blood smear shows large numbers of heavily stained reticulo. A properly prepared blood smear is essential to accurate assessment of cellular morphology. Blood smears should be prepared using wellmixed edtaanticoagulated blood. A few drops of blood were used to prepare blood films, and the. The test provides information on the number and shape of these. 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. An unconventional way to make peripheral blood smears duration.
Blood smears are needed for microscopic examination of the blood. Prepare a thick blood smear on a clean and dry microscopic glass slide and air dry it. Blood smears, at least two thick and two thin, should be prepared as soon as possible after collection. May 01, 2015 peripheral blood smear examination macroscopic view. A wellmade blood smear is a beauty to behold, and likely to yield interesting and. A simplified wrights stain technique for routine avian blood.
It is good practice to prepare two slides for each sample. A brief description of blood collection and handling is presented, followed by basic principles of haematological evaluation, including blood smear preparation and evaluation, and manual and. Preparation of human blood smear blood cells can be identified on the basis of the staining properties of the nucleus and cytoplasm. Do not touch the surface of the slide where the blood smear will be made. The blood droplet on the right is the preferred size. Delay in preparation of the smears can result in changes in parasite morphology.
To this end, this video describes in a virtual way the realization of a blood smear and the staining of its cellular components with giemsa staining. Too large thick and long smears too small thin and short smear ideal size of the drop id lentil shape angel. The manual wedge technique is used most commonly in our. The smear obtained closest to the date of diagnosis was used for each patient. Blood filled capillary tube or purple top with diffsafe dispenser. Over 1,000 fullcolor photomicrographs depict abnormalities within each blood cell line, with multiple pictures of each morphologic abnormality and variations in their appearance. The purpose of preparation of blood smear blood film is to study the morphology of rbcs, differential leukocyte count and reticulocyte count.
Always start with room temperature, wellmixed, clot free, edta anticoagulated blood heparin is used for some exotic species. Useful smears are shown in figure 3, a and b, even though one of the smears fig. Blood smears are useful for examining cell morphology and identifying blood borne parasites. Clean, highquality glass microscope slides are essential to blood smear preparation. Glass slides for blood film preparation should be new or precleaned and of high quality, and have a frosted edge for labeling. Preparation and staining of blood smears flashcards. Select two glass slides that are clean and free of chipped edges. The smear is stained with wrights stain and performed after the complete blood count is run. Preparation of blood smear with different staining method dr.
Blood is composed of the blood cells which accounts for 45% of the blood tissue by volume, with the remaining 55% of the volume composed of plasma, the liquid portion of the blood. For more information on how to make a good blood smear, please also refer to related links below. The volume of blood and the size of the thick lm detailed in the procedure below have been calculated based on. The optimal blood smear is a single layer of dispersed cells that accurately represents the distribution of cells in whole blood. Cytology reference guide dechra veterinary products. Apr 21, 2016 this video demonstrates the process of making a blood smear for observing the various corpuscles. Putting two smears per slide saves on weight glass is heavy for field trips. Key words artifacts, blood smear preparation, blood smear staining, peripheral. Peripheral blood smear image examination is a part of the routine work of every laboratory. A suitably sized droplet and a steady motion with the top spreading slide are essential for good blood film preparation, creating a defined and extensive feathered area. Blood smear prepared for you by an experienced technician smear evaluation performed by a technician on every specimen. If the blood has been refrigerated, allow it to return. When first learning, try multiple methods to determine which process works best for you.
Prior to staining, fix the material to be observed over the slide. The method is very easy and modern research must combine studies of. Alternately, edta anticoagulated blood may be used if the smear is made within one. Blood smears are made using fresh blood or edta blood. Place a small drop of blood, or one side about 12 cm from one end.
Without delay place a spreader at an angle of 45 from the slide and move it back to make contact with the drop. Learn vocabulary, terms, and more with flashcards, games, and other study tools. Preparation of peripheral blood smear and staining procedure. Making a quality blood smear although there are several techniques described for making blood smears, most people use the wedge or push technique. Introduction to peripheral blood smear examination. Capillary blood should be obtained by fingerstick, or venous blood should be obtained by venipuncture. This has motivated researchers to develop different algorithms and methods to automate peripheral blood smear image analysis. To achieve the appropriate sample volume for spreading see step 2, using a microhematocrit capillary tube rather than a pipette to place a drop of the sample on the glass slide is recommended. In 15 patients, a second archived specimen was also evaluated to determine the variability of the percentage of smudge cells over time. An extensive study was undertaken to compare five methods of blood smear preparation. The most reliable and complete diagnosis of blood smear infection is done by manually finding disorders and abnormalities in blood samples through a microscope, and counting blood smear particles.
A blood film or peripheral blood smear is 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. 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. Certain cell types can be damaged easily by the blood. The blood smears were prepared from either edta anticoagulated blood or a finger stick. Each person may have their own preferencemethods for success. Automated slide preparation and interpretation can enhance lab. Choose from 500 different sets of blood smear flashcards on quizlet. Using smudge cells on routine blood smears to predict. Remember to include a detailed history and description of. There are multiple methods to blood smear preparation.
A blood smear is a blood test used to look for abnormalities in blood cells. This new methylene bluestained blood smear shows large. Learn blood smear with free interactive flashcards. Estimation of platelet count in unstained peripheral blood. Diagnostic blood smear preparation clinicians brief. Choose from 500 different sets of preparing staining a blood smear flashcards on quizlet. 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. Making and staining a blood smear university of vermont. Clean slides with 70 to 90% alcohol and allow to dry. This procedure is very easy to perform and is suitable for routine avian hematology. The smear is greater than 25 mm long and the feathered edge stops approximately 10 mm from the end of the slide. Preparation of blood smears in order to reduce the number of unacceptable smears received, marshfield labs recommends the following procedure to be used in preparation of blood smears. The use of glass slides for desi profiling of blood samples leaves the opportunity to combine it with.
To assist your office in preparing the smears from the fresh blood in the needle tip, may we offer the following suggestions. Blood smear preparation properly prepared blood smears are essential to accurately assess the patients status. Introduction to peripheral blood smear examination oncohema key. Present blood smear to your instructor for evaluation. Seed haematology the role of the peripheral blood smear in. Update on malaria diagnostics and test utilization journal of. Blood smears can be made from the blood remaining in the needle by pushing a small drop of blood onto the slide with the last evacuated tube. Many hematological diagnoses depend upon this procedure and often a definitive diagnosis can be established from the careful examination of the blood film.
Use dull scalpel blade or medical spatula to collect samples from under a crust, surface of exudative or seborrheic skin lesion or a claw fold. Comparison of five methods for preparing blood smears. Video is prepared by the faculty and students of the department. Blood smears are used to determine leukocyte differentials, to evaluate erythrocyte, platelet and leukocyte morphology, and, if necessary, to estimate platelet and leukocyte counts.
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