Preparation of a peripheral blood film this protocol describes how to prepare a slide film of peripheral blood for microscopic examination. A phlebotomist, a person specifically trained to draw blood, first cleans and sterilizes the injection site with an antiseptic. Find a good area of the slide then scan at 25x or 50x examining cells at 100x as needed. The volume of blood and the size of the thick lm detailed in the procedure below have been calculated based on. Many hematological diagnoses depend upon this procedure and often a definitive diagnosis can be established from the careful examination of the blood film.
Examination of thin blood films is important in the investigation and management of anaemia, infections, and. The sample can be mixed by gently inverting the tube several times before slide preparation. 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. Place the airdried smear on the slide staining rack, smear side facing upwards. 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. 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.
Concentrating cells via a buffy coat preparation or performing. These are the same cases that you studied in the two labs the normal smear is in there as well. One requires slides, pipettecapillary tube and blood spreader to make pbf smear. Peripheral blood smear, preparation, examination, interpretation, reporting, blood cells morphology. The thin blood film should be thick enough at the starting point 1, becoming less thick in the.
Blood smear evaluation can help confirm platelet and white. This way, each slide may be used to spread the film of the other. It is good practice to prepare two slides for each sample. Preparation thin blood films and giemsa staining lqm laboratory.
Blood smear preparation properly prepared blood smears are essential to accurately assess the patients status. Do not touch the surface of the slide where the blood smear will be made. They then tie a band above the venous site where your blood will be drawn. Blood smears are needed for microscopic examination of the blood. A wellprepared blood smear can be a tremendous asset to the intrepid clinician. Blood smear evaluation can help confirm platelet and white blood cell. There are multiple methods to blood smear preparation. Alternately, edta anticoagulated blood may be used if the smear is made within one. Note that the diameter of the red blood cell is similar to the diameter of the small lymphocyte nucleus. Most commonly blood smears are used for the differential leukocyte count dlc.
The thick blood films are used for microscopic detection of plasmodium parasites, the causal. Preparation thin blood films and giemsa staining projectstudy. The bottom row displays poorly prepared blood smears. A properly prepared blood smear is essential to accurate assessment of cellular morphology. This made it possible to study individual blood cells by light microscopy after a small amount of blood had been placed and smeared onto glass slides, dried, fixed.
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. Freshly prepared blood smears should be submitted with all edta blood for cbc tests. If the smear is made too thin, there is a chance the larger cells will be collected. When carrying out a complete blood count, preparation and evaluation of a blood smear should always be performed alongside. Label precleaned slides preferably frostedend with patients name or other identifier, date and time of collection. 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. When first learning, try multiple methods to determine which process works best for you. The preparation of standard lms is aided by the use of slide templates that indicate the placement of the lms and their size. 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.
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 top two slides are prime examples of proper blood smears. To assist your office in preparing the smears from the fresh blood in the needle tip, may we offer the following suggestions. Slides should not be left uncovered on top of the counter. The undiluted stain fixes and partially stains the smear. Preparation of thick and thin blood smear the direct microscopic visualization of the malarial parasite on the thick andor thin blood smears has been the gold standard for malaria diagnosis. 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. Place a small drop of blood, or one side about 12 cm from one end. A carefully prepared blood smear is vital for an accurate differential count. Smears made from edtaanticoagulated blood should be made. The examination of the peripheral blood smear is an important basic hematological procedure.
White blood cells or leukocytes are primarily involved in defense from invasion and infection. Even the most uptodate hematology analyzers are no match for a diagnostic hematologist. Peripheral blood or potassium edta anticoagulated blood 12 mg edta1 ml blood may be used. A preparation of blood smear selection of a spreader. Smears must be thin, evenly fading toward the middle of the slide, leaving a feathered edge. Without delay place a spreader at an angle of 45 from the slide and move it back to make contact with the drop. 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. Take another clean slide, and holding at an angle of about 45. The classic blood smear has a thumb print appearance. An introduction to haematology and blood smear preparation. Blood smears should be prepared using wellmixed edtaanticoagulated blood. 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.
Blood smear prepared for you by an experienced technician smear evaluation performed by a technician on every specimen. This procedure describes the preparation of thin blood films. Select two glass slides that are clean and free of chipped edges. 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.
Certain cell types can be damaged easily by the blood. The smear is stained with wrights stain and performed after the complete blood count is run. 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. A fresh, wellmade, peripheral blood film is crucial for accurate cell morphology assessment. Warm patients hand with warm water, or by rubbing together briskly.
Using the ground edge of the spreader slide spread the blood for the thin film. Blood films that are too thin or too thick present a problem. Making the blood smear before any stained smear can be used for a diagnosis. Preparation of thick blood films and giemsa staining. 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. Preparation of blood smear with different staining method dr. A wellmade blood smear is a beauty to behold, and likely to yield interesting and. 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 peripheral blood film remains a very useful tool in hematological assessment. Diagnostic blood smear preparation clinicians brief. Disinfect tip of middle or ring finger with a 70% alcohol pad, and allow to dry completely. Norgan ap, arguello he, sloan lm, fernholz ec, pritt bs. 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. Examination of a blood smear is an integral part of a hemogram.
This new methylene bluestained blood smear shows large numbers of heavily stained reticulo. Slide preparation can be quite laborious especially if. It should be done by careful look on the narrow edge of the slide or by moving a thumb smoothly on its edge. 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. Blood lab first week this is a self propelled powerpoint study atlas of blood cells encountered in examination of the peripheral blood smear. Introduction to peripheral blood smear examination. Blood smears are made using fresh blood or edta blood. Thin smears consist of blood spread in a layer such that the thickness decreases progressively toward the feathered edge. Prepare a film of blood or bone marrow on a microscopic slide and allow to air dry. Each person may have their own preference methods for success. Scope and application this procedure describes the preparation of thick blood films.
Thick blood smear showing trypanosoma species staining the thin and thick blood smear on the same slide with giemsa stain. Clean slides with 70 to 90% alcohol and allow to dry. Preparation of thick blood films and giemsa staining projectstudy. Cover the blood film with undiluted staining solution. Blood smears can be examined by trained veterinary technicians, veterinarians, or sent to a diagnostic laboratory in conjunction with whole. Prepare a second smear by repeating steps 35, using the edge of the. Pdf the peripheral blood film pbf is a laboratory workup that involves cytology of. 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. 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. Blood filled capillary tube or purple top with diffsafe dispenser. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas.
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