It is now more common to have blood analyzed by a computer. How i tricked my brain to like doing hard things dopamine detox duration. The blood smear should occupy the central portion of the slide and should not touch the edges. To make a good blood smear, very little equipment is needed aside from practice in how to create a good smear and a good microscope. Fixation of the thin smear was done in a covered staining jar containing anhydrous methanol for 1 to 2 min, after which the slides were airdried. Procedure for thick and thin blood film staining technique. If aequous staining procedures will be used fix blood smear 3 min in cold meoh and dry 10 min on the air before proceeding. The spun smear requires an automatic slide spinner. Most lymphocytes are not much larger than the red blood cells, though their diameter may vary as much as 8 m to 12 m. Examination of a blood smear is an integral part of a hemogram.
A thin film was made by placing a small drop of blood on the centre of a greasefree microscopic slide. Blood smear preparation properly prepared blood smears are essential to accurately assess the patients status. Choose from 500 different sets of blood smear staining flashcards on quizlet. Simple stain individual supplies heatfixed slides see step i above methylene blue dye staining tray. For valuble information such as general health, wbc estimate, platelet estimate, relative proportions of different types of wbcs, cell morphology 2. Draw the spreader back, until it touches the drop of blood with the edge of the slide. Method for inactivating and fixing unstained smear. A blood smear is checked by a lab scientist or healthcare provider who specializes in blood or infectious diseases. Time spent learning and practicing to make good smears will be well worthwhile. The key element in staining was prewetting the fixed blood smear with the 0. Alternately, edta anticoagulated blood may be used if the smear is made within one. The stock buffer should be kept in the refrigerator, but if not possible, can be stored at room temperature for several weeks.
After staining with giemsa, the slide can be examined under the microscope. Note that the diameter of the red blood cell is similar to the diameter of the small lymphocyte nucleus. 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. L edta blood was then used for the simultaneous preparation of two thin and thick smear slides, one stained according to giemsa and the other according to leishmans method. A blood smear can also detect parasites in your blood. 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.
Once the blood smear is made, let it dry it will take a few minutes. Purpose and criteria for blood smear scan, blood smear. Make working buffer which can be stored at room temperature for a few days. Blood smears common errors the blood droplet on the left is too big and will result in a thick smear. 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.
Without delay, the blood was spread with a glass spreader held at a steep angle to achieve a thick smear covering an area of 15 x 15mm. Leave the slide to air dry and make more smears if required. This procedure describes the preparation of thick blood films. Prepare a film of blood or bone marrow on a microscopic slide and allow to air dry. 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 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. Blood smears are used to determine leukocyte differentials, to evaluate erythrocyte, platelet and leukocyte morphology, and, if necessary, to estimate platelet and leukocyte counts.
The blood can be obtained from finger heal microhematochrit tubes non heparinised for edta or heparinised for capillary blood wooden stick or glass capillary pipette. Staining for 3 minutes gave equally intense staining of the white blood cells. The stain needs adequate time with the sample to avoid over staining. This occurs when there is inadequate contact with the stain solutions. Place the airdried smear on the slide staining rack, smear side facing upwards. Dried blood smears may be stored at rt for several days before use. Then write the patients name in pencil on the bottom edge of the slide, as illustrated in figure 718, view d. Blood smear in microscopy processtechnique, artifacts. Comparing leishman and giemsa staining for the assessment. The dip method is a quick staining method that uses a modified wright giemsa stain buffered in methanol at ph 6.
Slides are immersed in the stain in a coplin jar for a user determined length of time. Using the corner of a clean slide, spread the drop of blood in a circle the size of a dime diameter 12 cm. Lymphocytes about 30% of all leukocytes all look alike in ordinary blood smear. Fill a capillary tube threequarter full with the anticoagulated specimen or a wooden stick. 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. The smear and simple stains san diego miramar college. 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. The inactivation of smears that contain mycobacterium tuberculosis for microscopy before removal of the material from a biosafety cabinet is an important safety factor in preventing the potential transmission of tuberculosis to laboratory workers.
The other is pusher or the spreader slide drop of blood 23 mm in diameter. Although this is a higher ph than normally used to stain blood cells, the. Do not make the smear too thick or it will fall off the slide. Our wrights stain can be used to stain blood smears in. You should make at least one smear from a solid inoculum and one smear from a liquid inoculum. Cover the blood film with undiluted staining solution. Add leismans stain drop by drop till it covers whole of the smear. Method of staining flood the smear with methylene blue, allow for 2 minutes, pour off the stain and allow the air to dry by keeping in a slanting position and by this the organism will retain the methylene blue stain use methylene blue staining is used to make out clearly the morphology of the organisms eg. Due to a lack of guidance documents, many organizers are reluctant to offer a. Introduction to peripheral blood smear examination. The blood smear is primarily ordered as a followup test when a cbc with differential, performed with an automated blood cell counter, indicates the presence of atypical, abnormal, or immature cells. 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. Place the edge of the spreader just in front of the drop of blood. Below is a stepbystep guide for veterinarians produced by the department of agriculture and food, western australia for performing blood smears for diagnostic use.
Fixation, staining, washing and air drying are quickly commenced. Prior to staining, if the slide is held up to the light, there is a rainbow effect seen just behind the feathered edge on well made smears. The thickness of the spread when pulling the smear is determined by the 1angle of the spreader slide the greater the angle, the thicker and shorter the smear. Preparation and staining of peripheral blood smear slideshare uses cookies to improve functionality and performance, and to provide you with relevant advertising. Large blood drops may extend the smear over too much of the base slide, while a small drop can be insufficient for the smear. Start a free trial of quizlet plus by thanksgiving lock in 50% off all year try it free. 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 following points highlight the two methods of staining of supplied smears. A smear with wavy lines or blanks spots should be discarded, and a new smear made. The stain contains a fixative, methanol, and the stain in one solution. The mechanism of wrights staining is a proc ess in which both chemical and physical factors playa role. The blood droplet on the right is the preferred size. A blood smear is a drop of blood spread thinly onto a glass slide that is then treated with a special stain and the blood cells on the slide are examined and evaluated. Jan 27, 2017 megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation.
Blood smear definition of blood smear by medical dictionary. Megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation. This was allowed to air dry in a horizontal position. Since good quality control smears are not available commercially, they may be prepared from a patients blood and stored for future use in the following manner. Terms in this set 30 importance of wellmade blood smear. Preparation of blood smear with different staining method dr. Staining blood smears with dilutions of 10x or 20x in either water, saline, dilute sodium bicarbonate or vitamin c only gave faint blue staining of white blood cells after 3.
Place the slide on a staining rackensuring that the smear is upper most. A blood smear is checked by a lab scientist or healthcare provider who specializes in. Due to students initial inexperience, slides are frequently broken and blood smears are damaged in microscopy training, leading to the need for their constant replacement. Histology laboratory microscope orientation and blood. Dried and fixed peripheral blood smears can easily be differentially stained with. Blood smear technique for veterinarians agriculture and food. Below is a very quick video on blood smear technique. Preparation of thick blood films and giemsa staining lqm.
Guidelines for blood smear preparation and staining procedure for. Proceeding with the 45 degree wedge or push slide technique used in manual and automated environments, creates a monolayer blood smear. If using a needle and syringe, first remove the needle and then touch the end of the syringe to the slide. Erythrocytes red blood cells, leukocytes white blood cells, and platelets can be easily identified using this slide. A properly prepared blood smear is essential to accurate assessment of cellular morphology. Comparing leishman and giemsa staining for the assessment of. Practical of clinical hematology islamic university of gaza.
Ahl will do this under controlled conditions to optimise the staining process. To ensure that proper staining results have been achieved, a positive smear malaria should be included with each new batch of working giemsa stain. Poor quality slides are typically cause by excessive downward pressure, a slow spreading motion, or. Learn vocabulary, terms, and more with flashcards, games, and other study tools. Usually examinee is provided with a fixed smear prepared from c. A rarer type of white blood cell, the basophil, was not seen or identified. 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 a wellmade blood smear is a beauty to behold, and likely to yield interesting and significant information for a research project. A phlebotomist, a person specifically trained to draw blood, first cleans and sterilizes the injection site with an antiseptic. They have a very darkly staining, dense, round nucleus. Feb 21, 20 how i tricked my brain to like doing hard things dopamine detox duration. It is important that the wrightgiemsa stain be kept tightly sealed in coplin jar when not in use and be replaced when water artifact. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas.
Then a giemsa stain 3% solution diluted with buffer for 30 minutes staining was applied on the thick film and allowed for 30 minutes. An optimal smear is threequarters the length of the slide and has a feathered edge. A well prepared blood smear is necessary for microscopic examination of blood. The fixing and inactivating properties of heat flaming, 70% ethanol, and 1, 3, and 5% phenol in ethanol for smears containing m. For the purpose of this lab exercise, we will use the wedge smear. The undiluted stain fixes and partially stains the smear. But blood smears are still routinely done to look for certain diseases. The stain needs adequate time with the sample to avoid overstaining. A blood smear scan serves to at least a verify the flagged automated hematology results and b determine if a manual differential leukocyte count needs to be performed.
The peripheral blood film remains a very useful tool in hematological assessment. Learn blood smear staining with free interactive flashcards. It may also be performed when a person has signs and symptoms that suggest a condition affecting blood cell production or lifespan. B staining of blood smear leave it for 12 minutes for fixation of the smear. To minimize this problem a method of preparing blood smears on transparent acetate sheets was developed with the goal of implementing appropriate and more readily available teaching resources for the microscopic. Most starting external quality organizers set up a scheme for clinical chemistry. Traditionally, trained laboratorians have examined blood smears manually using a microscope. Making and staining a blood smear university of vermont. A properly performed and analyzed blood smear is the most informative of all hematologic tests, allowing examination of erythrocytes. The scratches allow for improved adherence of the blood film to the slide without affecting the smear morphology. Introduction to peripheral blood smear examination clinical. However, the red blood cells took on more stain too, appearing darker.
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