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. Microscopy of peripheral blood thin and thick films remains the reference for malaria diagnosis. Comparison of five methods for preparing blood smears. 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. Icsh has published a reference staining method for blood films based on purified azure b and eosin y solutions ichs, 2010. 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. Stain only one set of smears, and leave the duplicates unstained. Too large thick and long smears too small thin and short smear ideal size of the drop id lentil shape angel. Aug 30, 2012 these findings suggested cryoglobulins.
A simple single differential stain of blood smears using. Stains may be used to define biological tissues highlighting. Megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation. Staining for 3 minutes gave equally intense staining of the white blood cells.
The proliferation of inclinic analyzers facilitates rapid turnaround time that can improve patient care. A series of five slides was prepared from each sample of blood. Oct 18, 2016 preparation of smear there are three types of blood smears. 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. The best place to examine blood cell morphology is the feathered edge of the blood smear where red cells lie in a single layer, side by side, just barely touching one another but not overlapping. It is hoped that this paper will provide an aid to those who wish to employ the blood smear as a diagnostic tool. Factors that influence making a blood smear size of the drop. The purpose of preparation of blood smear blood film is to study the morphology of rbcs, differential leukocyte count and reticulocyte count. Too small a drop of blood will result in too short a smear. Resident physician, cook county hospital chicago the value of blood smears to the practicingphysicianthe most importantfeature in all blood examinations is the stained blood smear. 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 thin smear of blood was stained using giemsa stain himedia, mumbai. The procedure of giemsa staining varies as per the purpose of staining that means whether the staining is done for the examination of blood cells or to find the parasites in the blood smear and accordingly the blood smears are prepared as thin blood films or thick blood films. 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. B staining of blood smear leave it for 12 minutes for fixation of the smear. Prior to staining, fix the material to be observed over the slide. 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. Blood films with excessive tails or gritty feathered ends indicate a spreader edge that is rough or dirty, or an accumulation of leukocytes due to either slow spreading or a very high leukocyte count. 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. Histology laboratory microscope orientation and blood smear lab for practicing how to use the microscope do not use the blood smear slide it is too boring for the lower mags. To ensure that proper staining results have been achieved, a positive smear malaria should be included with each new batch of working giemsa stain. The dense body of the smear takes up most of the slide and there is a thin counting area which blends with the feathered edge and the dense body. It is then treated with a special stain and studied under a microscope to examine the blood cells.
A peripheral blood smear examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide. The latter will prove useful if a problem occurs during the staining andor if you wish later to send the smears to a reference laboratory. Or the specimen being stained may be used as the organisms andor the white blood cells are a built in quality control. Either should be tested for proper staining reaction before use. The word gram is always spelled with a capital, referring to hans christian gram, the inventor of gram staining gram staining principles gram staining is used to determine gram status to classify bacteria broadly. Learn preparing staining a blood smear with free interactive flashcards. Choose from 500 different sets of preparing staining a blood smear flashcards on quizlet. Quality control to ensure that proper staining results have been achieved, if a positive smear malaria is available it may be included with each new batch of working giemsa stain. However, the red blood cells took on more stain too, appearing darker. After one minute staining with black quink, the walls of the red blood cells erythrocytes. 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. A properly performed and analyzed blood smear is the most informative of all hematologic tests, allowing examination of erythrocytes. Blood film preparation and staining procedures researchgate. Here, the methods for making and staining smears are given, as well as a list of sources for high quality slides, stain, and chemicals.
Right blood as seen at 400x under light microscope. Pdf when carrying out a complete blood count, preparation and evaluation of a blood smear should always be performed alongside. This procedure describes the preparation of thin blood films. The decolourising process is then stopped by immersing the slide in running tap water. The three main blood cells that the test focuses on are. Instructor in physiology, college of physicians and surgeons. 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. Although giemsa staining is most commonly used, the leishman staining method provides better visualization of the nuclear chromatin pattern of cells. Or the specimen being stained may be used as the organisms andor the white blood cells are a. Over 1,000 fullcolor photomicrographs depict abnormalities within each blood cell line, with multiple pictures of each morphologic abnormality and. Learn vocabulary, terms, and more with flashcards, games, and other study tools. Guidelines for blood smear preparation and staining procedure for. As a general rule for judging the quality of a stained blood film, the laboratory must ensure that all cell types in a blood film can be identified reliably by the staining procedure. Staining is a technique used to enhance contrast in samples, generally at the microscopic level.
Module common staining technique microbiology 24 notes indeterminate groups as well. Used in hematology, this stain is not optimal for blood parasites. A second romanowsky stain is therefore often used in combination with giemsa and contains azure dyes to intensify the staining of nuclear features. A blood smear is a blood test used to look for abnormalities in blood cells. This test uses a drop of blood spread onto a glass microscope slide that is then treated with a colored stain and examined using a microscope. The stain must be diluted for use with water buffered to ph 6. Smears made from edtaanticoagulated blood should be made. A blood smear is used to diagnose andor monitor numerous conditions including appropriate number and morphology of white blood cells wbcs, red blood cells rbcs, andor platelets plt. 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 chief executive officer of the department of agriculture and food and the state of western australia accept no liability whatsoever by reason of negligence or otherwise arising from the use or release of this information or any part of it. Preparation of blood smear with different staining method dr. In this lesson, readers will learn about the procedure, staining methods, and interpretation of the blood smear test. 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. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas.
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. It is less well known whether accuracy of parasitaemia assessment is equally accurate with the latter method. The smear is greater than 25 mm long and the feathered edge stops approximately 10 mm from the end of the slide. The other is pusher or the spreader slide drop of blood 23 mm in diameter. Peripheral blood smear can be used for est imation of. Introduction to peripheral blood smear examination.
An extensive study was undertaken to compare five methods of blood smear preparation. Stains and dyes are frequently used in histology the study of tissue under the microscope and in the medical fields of histopathology, hematology, and cytopathology that focus on the study and diagnoses disease at a microscopic level. Giemsa, a commonly used stain does not adequately stain red blood cells, platelets or white blood cell cytoplasms when used alone. Three mechanical devices for preparing blood smears were compared to the traditional coverslip and manual wedge methods.
The blood droplet on the left is too big and will result in a thick smear. Procedure of regressive papanicolaou staining method. The most dependable stain for blood parasites, particularly in thick films, is giemsa stain containing azure b. Start a free trial of quizlet plus by thanksgiving lock in 50% off all year try it free. Add leismans stain drop by drop till it covers whole of the smear. Preparation thin blood films and giemsa staining lqm laboratory. In the present study, out of total 218 cases tested, 19. This picture illustrates the proper procedure for making blood smears. Jul 15, 2019 a peripheral blood smear examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide. A properly prepared blood smear is essential to accurate assessment of cellular morphology. Making the blood smear before any stained smear can be used for a diagnosis. 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. Seed haematology the role of the peripheral blood smear.
Procedure place a drop of blood in the centre of a glass slide. Prior to staining, cells must be fixed to the glass slide with acetone free methanol, either alone or in solution with. The blood smear should occupy the central portion of the slide and should not touch the edges. Bloodfilmpreparationand stainingprocedures berendhouwen,md,phd. A serum prepared at 37c demonstrated protein precipitation when cooled to room temperature. Purpose and criteria for blood smear scan, blood smear. Blood smear definition of blood smear by medical dictionary. The mechanism of action of wrights stain is also discussed.
Seed haematology the role of the peripheral blood smear in. 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. Thin blood films are used differential leucocyte count, the detection of abnormal erythrocytes. Histology laboratory microscope orientation and blood. Spin at a high speed in a special centrifuge, cytospin. 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 bloodsmears by this method to help in differentiating typhoid, malaria, and.
Use of giemsa stain is the recommended and most reliable procedure for staining thick and thin blood films. Jan 27, 2017 megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation. Assign to person add to training plan add to bookmarks bookmarking is a premium feature. 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. Making and staining a blood smear university of vermont. Cryoglobulin deposits on a blood smear blood american. The excess stain is removed with dilute hydrochloric acid solution acid water.
If preparation is not fixed or sticked over the slide, the cells are washed off during staining procedure. The thickness of the spread when pulling the smear is determined by the 1 angle of the spreader slide the greater the angle, the thicker and shorter the smear, 2 size of the blood drop and 3 speed of spreading. The blood droplet on the right is the preferred size. The authors approach is to look at the smallest cellular elements, the platelets, first and work. When using the regressive staining method, the nucleus is deliberately overstained with a nonacidified haematoxylin. Introduction to peripheral blood smear examination clinical. This article aims to be a very practical guidance document for external quality assessment organizers for the set up of blood smear schemes. This lesson also includes some general descriptions of conditions indicated by. The peripheral blood film remains a very useful tool in hematological assessment. The classic blood smear has a thumb print appearance. Blood from a patient with anaemia will have a lower. Causes and corrections too alkaline stain thick blood. Atlas of canine and feline peripheral blood smears vetbooks.
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