The blood droplet on the right is the preferred size. 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. 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. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas. Prior to staining, cells must be fixed to the glass slide with acetone free methanol, either alone or in solution with.
Wait until the thin and thick films are completely dry before staining. 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. Spin at a high speed in a special centrifuge, cytospin. 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. Too large thick and long smears too small thin and short smear ideal size of the drop id lentil shape angel. The authors approach is to look at the smallest cellular elements, the platelets, first and work. Seed haematology the role of the peripheral blood smear. 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. Histology laboratory microscope orientation and blood. Causes and corrections too alkaline stain thick blood.
Assign to person add to training plan add to bookmarks bookmarking is a premium feature. Over 1,000 fullcolor photomicrographs depict abnormalities within each blood cell line, with multiple pictures of each morphologic abnormality and. 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. Smears made from edtaanticoagulated blood should be made.
Thin blood films are used differential leucocyte count, the detection of abnormal erythrocytes. Too small a drop of blood will result in too short a smear. A series of five slides was prepared from each sample of blood. The stain must be diluted for use with water buffered to ph 6. Preparation thin blood films and giemsa staining lqm laboratory. Oct 18, 2016 preparation of smear there are three types of blood smears. Staining is a technique used to enhance contrast in samples, generally at the microscopic level. 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. Either should be tested for proper staining reaction before use. This article aims to be a very practical guidance document for external quality assessment organizers for the set up of blood smear schemes. Or the specimen being stained may be used as the organisms andor the white blood cells are a. Right blood as seen at 400x under light microscope.
Icsh has published a reference staining method for blood films based on purified azure b and eosin y solutions ichs, 2010. Instructor in physiology, college of physicians and surgeons. Procedure of regressive papanicolaou staining method. Giemsa, a commonly used stain does not adequately stain red blood cells, platelets or white blood cell cytoplasms when used alone. 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 most dependable stain for blood parasites, particularly in thick films, is giemsa stain containing azure b. The excess stain is removed with dilute hydrochloric acid solution acid water. Choose from 500 different sets of preparing staining a blood smear flashcards on quizlet. 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. Blood smear definition of blood smear by medical dictionary.
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. Learn preparing staining a blood smear with free interactive flashcards. A serum prepared at 37c demonstrated protein precipitation when cooled to room temperature. Megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation. Aug 30, 2012 these findings suggested cryoglobulins. Blood from a patient with anaemia will have a lower. 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.
A thin smear of blood was stained using giemsa stain himedia, mumbai. 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. 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 second romanowsky stain is therefore often used in combination with giemsa and contains azure dyes to intensify the staining of nuclear features. 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. Comparison of five methods for preparing blood smears. Cryoglobulin deposits on a blood smear blood american. Add leismans stain drop by drop till it covers whole of the smear. The decolourising process is then stopped by immersing the slide in running tap water.
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. In this lesson, readers will learn about the procedure, staining methods, and interpretation of the blood smear test. Introduction to peripheral blood smear examination. 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. 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. Or the specimen being stained may be used as the organisms andor the white blood cells are a built in quality control.
The three main blood cells that the test focuses on are. It is less well known whether accuracy of parasitaemia assessment is equally accurate with the latter method. 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. This procedure describes the preparation of thin blood films. However, the red blood cells took on more stain too, appearing darker. The smear is greater than 25 mm long and the feathered edge stops approximately 10 mm from the end of the slide. This picture illustrates the proper procedure for making blood smears. Factors that influence making a blood smear size of the drop. Three mechanical devices for preparing blood smears were compared to the traditional coverslip and manual wedge methods. Jan 27, 2017 megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation. Although giemsa staining is most commonly used, the leishman staining method provides better visualization of the nuclear chromatin pattern of cells. Atlas of canine and feline peripheral blood smears vetbooks. 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. Peripheral blood or potassium edta anticoagulated blood 12 mg edta1 ml blood may be used.
A simple single differential stain of blood smears using. Here, the methods for making and staining smears are given, as well as a list of sources for high quality slides, stain, and chemicals. Used in hematology, this stain is not optimal for blood parasites. The classic blood smear has a thumb print appearance. After one minute staining with black quink, the walls of the red blood cells erythrocytes. The blood smear should occupy the central portion of the slide and should not touch the edges. The mechanism of action of wrights stain is also discussed. Stain only one set of smears, and leave the duplicates unstained.
A peripheral blood smear examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide. It is hoped that this paper will provide an aid to those who wish to employ the blood smear as a diagnostic tool. 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. 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. 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. Making the blood smear before any stained smear can be used for a diagnosis. 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. Stains may be used to define biological tissues highlighting. Start a free trial of quizlet plus by thanksgiving lock in 50% off all year try it free. 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. Purpose and criteria for blood smear scan, blood smear.
Module common staining technique microbiology 24 notes indeterminate groups as well. Pdf when carrying out a complete blood count, preparation and evaluation of a blood smear should always be performed alongside. Guidelines for blood smear preparation and staining procedure for. Staining for 3 minutes gave equally intense staining of the white blood cells. 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. 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. Introduction to peripheral blood smear examination clinical. Blood film preparation and staining procedures researchgate. Bloodfilmpreparationand stainingprocedures berendhouwen,md,phd. Making and staining a blood smear university of vermont. 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. This lesson also includes some general descriptions of conditions indicated by.
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. Microscopy of peripheral blood thin and thick films remains the reference for malaria diagnosis. It is then treated with a special stain and studied under a microscope to examine the blood cells. A properly prepared blood smear is essential to accurate assessment of cellular morphology. Jul 15, 2019 a peripheral blood smear examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide.
The blood droplet on the left is too big and will result in a thick smear. 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. A properly performed and analyzed blood smear is the most informative of all hematologic tests, allowing examination of erythrocytes. B staining of blood smear leave it for 12 minutes for fixation of the smear. The peripheral blood film remains a very useful tool in hematological assessment. A blood smear is a blood test used to look for abnormalities in blood cells. An unstained blood smear is clearly seen by eye on the microscope slide, however, the blood cells are barely visible under the microscope figure 1. The proliferation of inclinic analyzers facilitates rapid turnaround time that can improve patient care.
Procedure place a drop of blood in the centre of a glass slide. 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. In the present study, out of total 218 cases tested, 19. Preparation of blood smear with different staining method dr. Learn vocabulary, terms, and more with flashcards, games, and other study tools. The wedge smear the cover glass smear the spun smear the are two additional types of blood smear used for specific purposes buffy coat smear thick blood smears for blood parasites 10. 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. 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 procedure. The purpose of preparation of blood smear blood film is to study the morphology of rbcs, differential leukocyte count and reticulocyte count. Prior to staining, fix the material to be observed over the slide. 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. When using the regressive staining method, the nucleus is deliberately overstained with a nonacidified haematoxylin.
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