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 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. Proper blood smear preparation texas veterinary medical. Even the most uptodate hematology analyzers are no match for a diagnostic hematologist. Blood smears, at least two thick and two thin, should be prepared as soon as possible after collection. Slides are just automatically dipped in the stain in the buffer and a series of rinses. Microscope slides preparation styles and techniques using prepared microscope slides. Hold top slide securely and pull back into entire drop of blood.
Atlas of canine and feline peripheral blood smears. Note that the diameter of the red blood cell is similar to the diameter of the small lymphocyte nucleus. This way, each slide may be used to spread the film of the other. 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. Peripheral blood or potassium edta anticoagulated blood 12 mg edta1 ml blood may be used. 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.
For more information on how to make a good blood smear, please also refer to related links below. A series of five slides was prepared from each sample of blood. As soon as the drop of blood is placed on the glass slide, the smear should be made without delay. A wellprepared blood smear can be a tremendous asset to the intrepid clinician. Apr 23, 2014 just recently i emailed my friend and said funny well, not really how many psychologists, doctors, dieticians, support groups, hospitals, and even a hypnotist, i went to over the years that couldnt help me and all it took in the end was a simple site like this, and a supportive partner. Smears must be thin, evenly fading toward the middle of the slide, leaving a feathered edge. Jun 12, 2016 1 introduction to peripheral blood smear examination. This picture illustrates the proper procedure for making blood smears. Label precleaned slides preferably frostedend with patients name or other identifier, date and time of collection. Guidelines for blood smear preparation and staining procedure. 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. Examination of thin blood films is important in the investigation and management of anaemia, infections, and. Most commonly blood smears are used for the differential leukocyte count dlc. The examination of the peripheral blood smear is an important basic hematological procedure.
Freshly prepared blood smears should be submitted with all edta blood for cbc tests. Introduction to peripheral blood smear examination oncohema key. Rouleaux formation blood american society of hematology. 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.
The three main blood cells that the test focuses on are. For a blood smear, the lab professional looks for blood cell problems that may not be seen on a computer analysis. Paul ehrlich introduced eosin as the first of these dyes for staining blood films in 1856, followed by hematoxylin in 1865, and later by the metachromatic. The other is pusher or the spreader slide drop of blood 23 mm in diameter. Set the second slide in front of blood drop at 30 to 45degree angle. Introduction to peripheral blood smear examination. This procedure describes the preparation of thin blood films. Do not touch the surface of the slide where the blood smear will be made. Find a good area of the slide then scan at 25x or 50x examining cells at 100x as needed. 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. Preparation thin blood films and giemsa staining lqm laboratory.
Take another clean slide, and holding at an angle of about 45. Slides should not be left uncovered on top of the counter. Blood smear definition of blood smear by medical dictionary. Place the airdried smear on the slide staining rack, smear side facing upwards. A properly performed and analyzed blood smear is the most informative of all hematologic tests, allowing examination of erythrocytes. The purpose of preparation of blood smear blood film is to study the morphology of rbcs, differential leukocyte count and reticulocyte count. Request pdf guidelines for blood smear preparation and staining procedure for setting up an external quality assessment scheme for blood smear. To illustrate the simplicity of the method of making and staining slide smears, let me say that the physicians in the receiving ward of the cook county hospital, working two at a time, and diagnosing from 100 to 200 cases daily, find time to make, stain and examine blood smears by this method to help in differentiating typhoid, malaria, and. Staining process has begun, no stat slides can be added in the batch. Preparation of a peripheral blood film this protocol describes how to prepare a slide film of peripheral blood for microscopic examination. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas. An extensive study was undertaken to compare five methods of blood smear preparation. Blood smear preparation requires two clean glass slides frosted at one end. A blood smear is a blood test used to look for abnormalities in blood cells.
Ideally, blood smear evaluation should be performed on a properly prepared blood smear to improve the assessment of all cell lines. 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. Choose from 500 different sets of preparing staining a blood smear flashcards on quizlet. How to perform a proper thick and thin smear for malaria diagnosis duration. Tutorial blood cell morphology louisiana state university health sciences center department of pathology new orleans, louisiana by carolyn sue walters, mhs, mtascp. Learn preparing staining a blood smear with free interactive flashcards. The peripheral blood film remains a very useful tool in hematological assessment. Concentrating cells via a buffy coat preparation or performing. Blood smears are needed for microscopic examination of the blood. Making and staining a blood smear university of vermont. Preparation and staining methods for blood and bone marrow. A fresh, wellmade, peripheral blood film is crucial for accurate cell morphology assessment. It takes about 510 minutes to stain a batch of smears.
Blood smears are used to determine leukocyte differentials, to evaluate erythrocyte, platelet and leukocyte morphology, and, if necessary, to estimate platelet and leukocyte counts. Objects magnified under compound microscopes are mounted onto microscope slides. These are the same cases that you studied in the two labs the normal smear is in there as well. Preparation of blood smear with different staining method dr. 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. This new methylene bluestained blood smear shows large numbers of heavily stained reticulo. The clinical pathology laboratory uses the wedge technique for preparation of blood smears. Place small drop of blood near frosted end of smear.
Over 1,000 fullcolor photomicrographs depict abnormalities within each blood cell line, with multiple pictures of each morphologic abnormality and variations in their appearance. 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. Pdf when carrying out a complete blood count, preparation and evaluation of a blood smear should always be performed alongside. Delay in preparation of the smears can result in changes in parasite morphology. The nucleus, which is acidic in character, has an affinity for basic stains and the. Filed under hematology, oncology and palliative medicine. Prepare a film of blood or bone marrow on a microscopic slide and allow to air dry.
Visual examination of blood smears is an essential tool for analysis, prevention, and remediation of several types of maladies. Deriving full value from blood smear examination requires a wellprepared, wellstained blood smear and some basic skills in the methods of assessment. Blood smear preparation properly prepared blood smears are essential to accurately assess the patients status. Request pdf guidelines for blood smear preparation and staining procedure for setting up an external quality assessment scheme for blood smear interpretation. One requires slides, pipettecapillary tube and blood spreader to make pbf smear. Apr 21, 2016 this video demonstrates the process of making a blood smear for observing the various corpuscles. Comparison of five methods for preparing blood smears. The interest of computeraided decision has been acknowledged in. Glass slides should be free of chipped edges, glass fragments, dust, powder, fingerprints, and other debris, as these can affect blood spreading. Alternately, edta anticoagulated blood may be used if the smear is made within one. Three mechanical devices for preparing blood smears were compared to the traditional coverslip and manual wedge methods. If preparation is not fixed or sticked over the slide, the cells. The undiluted stain fixes and partially stains the smear.
Prior to staining, fix the material to be observed over the slide. In some ways it is becoming a lost art but it often provides rapid, reliable access to information about a variety of hematologic disorders. It is good practice to prepare two slides for each sample. Guidelines for blood smear preparation and staining procedure for. Prepare a second smear by repeating steps 35, using the edge of the. Thick blood smear showing trypanosoma species staining the thin and thick blood smear on the same slide with giemsa stain. Prepare a thin and thick smear of the specimen on the same slide which can be done by dividing the slides into two parts and then making the thin smear on one side and thick on the other side. Microscope slides preparation styles and techniques. Preparation and staining methods for blood and bone marrow films. An introduction to haematology and blood smear preparation. A properly prepared blood smear is essential to accurate assessment of cellular morphology. Blood smear evaluation can help confirm platelet and white.
Edta causes red blood cells to enlarge and lyse and will distort parasites making speciation difficult. Extremely thin films caused by too small a drop, too slow. Capillary blood should be obtained by fingerstick, or venous blood should be obtained by venipuncture. The value of the blood smear exam conference recording duration. In thin films the red blood cells are fixed and parasitized cells can be identified by.
Preparation of human blood smear blood cells can be identified on the basis of the staining properties of the nucleus and cytoplasm. Always mix the specimen immediately before making the smear by gently rolling the tube several times to ensure good cellular distribution, dont shake or invert. Clean, highquality glass microscope slides are essential to blood smear preparation. Aqueous solutions of stains are stable only after 36. Secondly, the pcv of the patient does influence the angle at which you hold the upper slide for a good smear. Examination of the peripheral blood smear is an inexpensive but powerful diagnostic tool in both children and adults. Introduction to peripheral blood smear examination clinical. Diagnostic blood smear preparation clinicians brief. White blood cells or leukocytes are primarily involved in defense from invasion and infection. A wellmade blood smear is a beauty to behold, and likely to yield interesting and.
This method produces a gradual decrease in thickness of the blood from thick to thin ends with the smear terminating in a feathered edge approximately 2 mm long. Dec 08, 2006 a well prepared blood smear is necessary for microscopic examination of blood. 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. Cover the blood film with undiluted staining solution. Accurate differential cell counts, cell morphology abnormalities, hemoparasites, neoplastic cells, and a host of other changes can be identified using an adequate blood smear. The appearance of rouleaux may be artificially caused by a poor preparation of the smear or by viewing the slide in a thickened area. The direct microscopic visualization of the malarial parasite on the thick andor thin blood smears has been the gold standard for malaria diagnosis. The smear is stained with wrights stain and performed after the complete blood count is run.
Many hematological diagnoses depend upon this procedure and often a definitive diagnosis can be established from the careful examination of the blood film. A blood filmor 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. Blood smear analysis allows quantitation of the different types of leukocytes called the differential count, estimation of the platelet count, and detection of morphologic abnormalities that may be indicators of pathophysiologic processes. Examination of a blood smear is an integral part of a hemogram. A couple of comments it would be beneficial if you showed a closeup of the finished slide so that viewers could appreciate what you describe. Peripheral blood smear test purpose, procedure, results. 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.
Preparation thin blood films and giemsa staining projectstudy. Made of glass or plastic, slides are approximately 1x3 inches and between 1mm1. Fields stain is a histological method for staining of blood smears. The definitive diagnosis of malaria infection and other parasites is still based on finding such organisms in blood films. Blood films that are too thin or too thick present a problem. Plasmodium ovale centers for disease control and prevention. Pdf feline blood smear preparation and evaluation researchgate. The smear is greater than 25 mm long and the feathered edge stops approximately 10 mm from the end of the slide. Smears made from edtaanticoagulated blood should be made. Blood smear preparation pdf download 3d39b66ab9 this material enables the simple and lowcost preparation of samples and does not compromise the morphologic and staining characteristics of the parasites or blood cellsthis ensured that the entire length of the smear on the acetate was simultaneously stainedor its licensors or contributorssecond. The thin blood films are prepared from whole blood with clean, dry and wrapped. Clean slides with 70 to 90% alcohol and allow to dry. 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.
Films made on coverglasses have negligible advantages. Handle a clean slide by its edge, label the target place at the bottom side o the slide by drawing a circle with a diameter about 2 cm using a marker. The stacking of cells rouleaux formation facilitates the rate of red cell sedimentation, a phenomenon that may be seen on a peripheral smear. Tutorial blood cell morphology a clinical pathology 201. 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. May 01, 2015 peripheral blood smear examination macroscopic view.