Prior to staining, fix the material to be observed over the slide. Most commonly blood smears are used for the differential leukocyte count dlc. The differential staining allows one to identify the types of white blood cells on the smear. Check out the leishman staining technique used in hematology laboratory for the examination of peripheral blood smear. Making a quality blood smear although there are several techniques described for making blood smears, most people use the wedge or push technique. The following article has been written to promote the greater use of slide bloodsmears in diagnosis by pointing out simple methods of making them. 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. Bloodsmeartest preparation fixation staining clinical diagnostic lukas k. Dip the thick blood smear into diluted giemsa stain prepared by taking 1ml of the stock solution and adding to 49ml of phosphate buffer or distilled water, but the results may vary differently. A peripheral blood smear or peripheral blood film is a glass slide covered on one side with a thin layer of venous blood.
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. Place the airdried smear on the slide staining rack. The hemopoietic stem cells produce white blood cells, red blood cells and platelets. The aim of the experiment is preparation and staining of bone marrow smear. The smear can then be stained as soon as it is dry, generally within 2030 minutes of smear preparation. Guidelines for blood smear preparation and staining. Comparing leishman and giemsa staining for the assessment of.
Cover the blood film with undiluted staining solution. Place the airdried smear on the slide staining rack, smear side facing upwards. Norgan ap, arguello he, sloan lm, fernholz ec, pritt bs. Blood film preparation methods for malaria detection. Leishmans stain, which occupies an intermediate position, is still widely used in the routine staining of blood films, although the results are inferior to those obtained by the combined maygrunwaldgiemsa, jennergiemsa, and azure beosin y methods. Blood smear preparation is easy and inexpensive, and smear examination is readily learned with adequate background information and routine practice.
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. Preparation and staining methods for blood and bone marrow. Request pdf guidelines for blood smear preparation and staining procedure for setting up an external quality assessment scheme for blood smear. Giemsa staining technique principle, preparation, procedure. 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. 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. A fresh, wellmade, peripheral blood film is crucial for accurate cell morphology assessment. Learn preparing staining a blood smear with free interactive flashcards. A thin smear of blood was stained using giemsa stain himedia, mumbai, india using standard protocol houwen, 2000. The interference of moisture in leishman stain is a wellknown problem in tropical climate and in rainy season, even in most wellequipped laboratories. A blood filmor peripheral blood smearis a thin layer of blood smeared on a glass microscope slide and then stained in such a. Appearance of an acceptable blood smear the perfect blood smear has a feathered edge that is nearly square, has a rainbow sheen when reflecting the light and is exactly one cell thick in the feathered edge when viewed microscopically. Guidelines for blood smear preparation and staining procedure for setting up an external quality assessment scheme for blood smear interpretation. Jul 25, 2019 prepare a film of blood or bone marrow on a microscopic slide and allow to air dry.
Blood smear definition of blood smear by medical dictionary. If there will be a delay in staining smears, dip the thick smear briefly in water to hemolyse the rbcs. Choose from 500 different sets of preparing staining a blood smear flashcards on quizlet. The purpose of preparation of blood smear blood film is to study the morphology of rbcs, differential leukocyte count and reticulocyte count. Quality control visually, the smear should appear as a round to oval smear of blood about 2 cm in diameter. Proper preparation of the blood smear is critical for obtaining accurate results on the differential. 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. In this lesson, readers will learn about the procedure, staining methods, and interpretation of the blood smear test. Preparation and staining methods for blood and bone marrow films. A properly prepared blood smear is essential to accurate assessment of cellular morphology. 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.
This lesson also includes some general descriptions of conditions indicated by. Introduction to peripheral blood smear examination clinical. A blood smear is a blood test used to look for abnormalities in blood cells. 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. The undiluted stain fixes and partially stains the smear. The progenitor cells mature into blood and lymphoid cells.
After staining with giemsa, the slide can be examined under the microscope. Wrights stain is a type of romanowsky stain, which is commonly used in hematology laboratory for the routine staining of peripheral blood smears. Feb 12, 2020 this video demonstrates the preparation of a thin peripheral blood smear and staining of the smear to study the various cellular components of blood. Before staining and observing a microbe under a microscope, a smear must be prepared. Blood smears are needed for microscopic examination of the blood. Blood films should be fixed in methanol within 4 hours preferably within 1 hour of preparation. Proper preparation of the blood smear is critical for correct interpretation and obtaining accurate results. This method is used to differentiate cells in the smear preparation of various gynecological specimens pap smears, materials containing exfoliative cells and material from fine needle aspiration. This drop of blood is then placed on a clean glass slide about 1 cm from one end in the center line of the slide.
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. If the methanol contains more than 3% water, morphologic artifacts including loss of cellular detail and vacuolation may be present. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas. Any delay results in an abnormal distribution of the white blood. The following article has been written to promote the greater use of slide blood smears in diagnosis by pointing out simple methods of making them. The purpose with this instruction is to describe the method used by boule for preparation and staining a blood smear and counting of the wbc subpopulations in a microscope. Blood films that are too thin or too thick present a problem. 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. Blood smear in microscopy processtechnique, artifacts. Certain cell types can be damaged easily by the blood. Thin smears consist of blood spread in a layer such that the thickness decreases progressively toward the feathered edge. Concentrating cells via a buffy coat preparation or performing. Wash the smear by dipping in in buffered water of distilled water for 35 minutes. Preparation, principle, procedure and results of wrights stain in hematology for staining peripheral blood smears and bone marrow aspirates.
Preparation and staining of bone marrow smear labmonk. For a blood smear, the lab professional looks for blood cell problems that may not be seen on a computer analysis. An equal volume of wrights buffer is added, and the solutions are mixed by blowing gently on the surface of the stainbuffer mixture. Dec, 2016 making and staining blood smears vet clinpath. Staining is a technique used to enhance contrast in samples, generally at the microscopic level. The bone marrow stroma has fibroblasts, macrophages, adipocytes, osteoblasts, osteoclasts, endothelial cells. It should be of such thickness that newsprint can barely be seen through the wet or dry smear.
To ensure that proper staining results have been achieved, a positive smear malaria should be included with each new batch of working giemsa stain. This chapter describes techniques of preparation and interpretation of canine and feline blood smears and addresses integrating findings from blood smears with other parameters in cbcs. The three main blood cells that the test focuses on are. Blood film preparation and staining procedures clinics in. It is incubated for 3 minutes to allow adequate fixation of blood cells and platelets.
Glass slides for blood film preparation should be new or precleaned and of high quality, and have a frosted edge for labeling. The student will prepare at least five slide smears which are even, smooth and have an acceptable feathered edge. Making and staining a blood smear university of vermont. Guidelines for blood smear preparation and staining procedure for. Blood film preparation and staining procedures researchgate. As soon as the drop of blood is placed on the glass slide, the smear should be made without delay. 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. Fixation, staining, washing and air drying are quickly commenced. Leishman staining technique principle, preparation. Introduction to peripheral blood smear examination.
A properly performed and analyzed blood smear is the most informative of all hematologic tests, allowing examination of erythrocytes. The examination of a stained peripheral blood film is a hematology test and is an integral part of laboratory evaluation of patients. How to stain peripheral blood and interpret blood film. In such a scenario, troubleshooting for suboptimal staining becomes challenging. Slide preparation for wbc differential by manual microscopy. Previously used or washed slides may be contaminated with residual detergents or microscopic debris that can appear as artifacts in the blood smear. Veterinary clinical examination of small animals and large animals. It is hoped that this paper will provide an aid to those who wish to employ the blood smear as a diagnostic tool. This video demonstrates the preparation of a thin peripheral blood smear and staining of the smear to study the various cellular components of blood. Dip fixed smear to field stain b red stain for 5 to 6 seconds.
P roceeding with the 45 degree wedge or push slide technique used in manual and automated environments, creates a monolayer blood smear. Learn vocabulary, terms, and more with flashcards, games, and other study tools. 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. Making the blood smear before any stained smear can be used for a diagnosis. Add a thick smear of blood and air dry for 1 hour on a staining rack.
Megan brashear, cvt, vts ecc, demonstrates technique in staining a blood smear for microscopic evaluation. 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. Nov 14, 2017 the art of peripheral blood smear preparation, staining, and examination is losing its shine. The smear is greater than 25 mm long and the feathered edge stops approximately 10 mm from the end of the slide. 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. Films made on coverglasses have negligible advantages. The thin blood films are prepared from whole blood with clean, dry and wrapped microscope slides cfr. Leishman stain is a mixture of methylene blue, and eosin dye, prepared in alcohol medium and diluted with buffer or distilled water during staining procedure. Blood smear preparation and staining mlab 15 hematology blood smear preparation and staining lab objective 1. A blood filmor peripheral blood smearis a thin layer of blood smeared on a glass microscope slide and then stained in such a way. The mechanism of action of wrights stain is also discussed. Peripheral blood smear procedure and interpretation. The blood smear is placed on a staining rack and flooded with wrights stain.
Films may be spread by hand or by automated slidspreader. The slide is the stained and viewed under a microscope. Giemsa stain principle, procedure, results, interpretation. Wrights stain is named for james homer wright, who devised the stain in 1902 based on a modification of romanowsky stain.
It is also used for staining bone marrow aspirates, urine samples and to demonstrate malarial parasites in blood smears. The student will stain two of the above smears with wrights stain so that all formed elements are readily identifiable according to. Using a sterilized inoculating loop, transfer loopful of liquid suspension containing bacteria to a slide clean grease free microscopic slide or transfer an isolated colony from a culture plate to a slide with a water drop. T hen plac e the blood drop 1cm from the end of the slide. 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. Preparation thin blood films and giemsa staining projectstudy. Oct 31, 2016 blood smear preparation, fixation, staining. A phlebotomist, a person specifically trained to draw blood, first cleans and sterilizes the injection site with an antiseptic.
Eosin y gives a pink colour to cytoplasm of mature squamous cells, nucleoli, cilia and red blood cells. The art of peripheral blood smear preparation, staining, and examination is losing its shine. Fix in methanol for one minute or get spray easyfix. Preparation of blood smear with different staining method dr. Ppt preparation, staining and examination of blood film. The main methods of placing samples onto microscope slides are wet mount, dry mount, smear, squash and staining. Make blood smear on a clean glass slide and it is dried in the air. The goal of smear preparation is to place an appropriate concentration of cells on a slide and then cement them there so that they do not wash off during the subsequent staining procedure.
This sample is sent to the laboratory for testing within two hours of collection. Extremely thin films caused by too small a drop, too slow. Microscope slides preparation styles and techniques. Diagnostic blood smear preparation clinicians brief. A wellmade blood smear is a beauty to behold, and likely to yield interesting and. 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. Filed under hematology, oncology and palliative medicine. Or the specimen being stained may be used as the organisms and or the white blood cells are a built in quality control. Preparation, staining and examination of blood film.
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