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. The three main blood cells that the test focuses on are. Learn preparing staining a blood smear with free interactive flashcards. 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. Or the specimen being stained may be used as the organisms andor the white blood cells are a. A blood smear is a blood test used to look for abnormalities in blood 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. A series of five slides was prepared from each sample of blood. Disperse the bacteria on the loop in the drop of water on the slide and spread the drop over an area the size of a dime. Place the airdried smear on the slide staining rack, smear side facing upwards.
Previously used or washed slides may be contaminated with residual detergents or microscopic debris that can appear as artifacts in the blood smear. Different types of staining methods are used to make the cells and. Too small a drop of blood will result in too short a smear. Preparation and staining methods for blood and bone marrow films. Cover the blood film with undiluted staining solution. This occurs when there is inadequate contact with the stain solutions. Heatfix the smear cautiously by passing the underside of the slide through the burner flame two or three times. Either should be tested for proper staining reaction before use.
Any delay results in an abnormal distribution of the white blood. Prior to staining, fix the material to be observed over the slide. Choose from 500 different sets of preparing staining a blood smear flashcards on quizlet. Slide preparation for wbc differential by manual microscopy. Comparison of five methods for preparing blood smears. The thick blood films are used for microscopic detection of plasmodium parasites, the causal. Microscope slides preparation styles and techniques. The most dependable stain for blood parasites, particularly in thick films, is giemsa stain containing azure b.
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. Things every technician should know clinical pathology susan kolkka, aas, lvt, vts the blood smear is an amazing thing. 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 lesson also includes some general descriptions of conditions indicated by. The progenitor cells mature into blood and lymphoid cells. Jul 25, 2019 prepare a film of blood or bone marrow on a microscopic slide and allow to air dry. Filed under hematology, oncology and palliative medicine. The aim of the experiment is preparation and staining of bone marrow smear. The cbc and the microscopic revision of blood smear are the mainstays of laboratory hematology. B staining of blood smear leave it for 12 minutes for fixation of the smear. The main methods of placing samples onto microscope slides are wet mount, dry mount, smear, squash and staining. One requires slides, pipettecapillary tube and blood spreader to make pbf smear. A variety of methods are available for preparing and staining blood smears, the most common of which are discussed in this atlas. The following article has been written to promote the greater use of slide bloodsmears in diagnosis by pointing out simple methods of making them.
Concentrating cells via a buffy coat preparation or performing. 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. Preparation of thick blood films and giemsa staining projectstudy. Scope and application 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. After staining with giemsa, the slide can be examined under the microscope. 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. Preparation, principle, procedure and results of wrights stain in hematology for staining peripheral blood smears and bone marrow aspirates. The undiluted stain fixes and partially stains the smear. Glass cover slips are mounted on all blood smears to prevent damage to smear during examination, cleaning, handling and storage.
Making the blood smear before any stained smear can be used for a diagnosis. Preparation and staining of bone marrow smear labmonk. Eosin y gives a pink colour to cytoplasm of mature squamous cells, nucleoli, cilia and red blood cells. Proper preparation of the blood smear is critical for obtaining accurate results on the differential. Peripheral blood or potassium edta anticoagulated blood 12 mg edta1 ml blood may be used. The mechanism of action of wrights stain is also discussed. 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 stain must be diluted for use with water buffered to ph 6. Guidelines for blood smear preparation and staining procedure for setting up an external quality assessment scheme for blood smear interpretation. In this lesson, readers will learn about the procedure, staining methods, and interpretation of the blood smear test. Cytohaematology is the study of the cellular components of blood cells. This way, each slide may be used to spread the film of the other, and duplicate slides will be of value if something happens to one of them in the.
Place the airdried smear on the slide staining rack. 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. 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. The smear is greater than 25 mm long and the feathered edge stops approximately 10 mm from the end of the slide. 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. This procedure describes the preparation of thin blood films. As technicians, we can have a vital role in the diagnosis and treatment of our. Introduction to peripheral blood smear examination. Glass slides for blood film preparation should be new or precleaned and of high quality, and have a frosted edge for labeling. Assign to person add to training plan add to bookmarks bookmarking is a.
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. 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. It is hoped that this paper will provide an aid to those who wish to employ the blood smear as a diagnostic tool. Preparation of human blood smear blood cells can be identified on the basis of the staining properties of the nucleus and cytoplasm. A phlebotomist, a person specifically trained to draw blood, first cleans and sterilizes the injection site with an antiseptic. How to stain peripheral blood and interpret blood film. 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. Glass slides should be free of chipped edges, glass fragments, dust, powder, fingerprints, and other debris, as these can affect blood spreading. The hemopoietic stem cells produce white blood cells, red blood cells and platelets. 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. Certain cell types can be damaged easily by the blood. Jan 12, 2020 this technique was developed by george papanicolaou, the father of cytopathology. Using the corner of a clean slide, spread the drop of blood in a circle the size of a dime diameter 12 cm. The classic blood smear has a thumb print appearance.
Megan brashear, cvt, vts ecc, demonstrates how to make a blood smear. Select two glass slides that are clean and free of chipped edges. Or the specimen being stained may be used as the organisms and or the white blood cells are a built in quality control. Three mechanical devices for preparing blood smears were compared to the traditional coverslip and manual wedge methods. It is good practice to prepare two slides for each sample. Most commonly blood smears are used for the differential leukocyte count dlc. Making and staining a blood smear university of vermont.
The thin blood films are prepared from whole blood with clean, dry and wrapped microscope slides cfr. Guidelines for blood smear preparation and staining procedure for. Diagnostic blood smear preparation clinicians brief. It is used to diagnose diseases, such as malaria, which are caused by parasitic infections the collected blood sample is smeared onto a glass slide and stained with giemsa stain. Learn vocabulary, terms, and more with flashcards, games, and other study tools. Operate this stain far from an open source of fire. Parasites may also be observed during microscopic examination of the blood smear. Add leismans stain drop by drop till it covers whole of the smear. As earlier mentioned,the examination of a stained peripheral smear is an integral part of laboratory evaluation of patients and this provides information on red cells, leukocytes and platelets and is used to supplement the information provided by automated hematology analyzers. 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.
Do not make the smear too thick or it will fall off the slide. Preparation thin blood films and giemsa staining projectstudy. Preparation thin blood films and giemsa staining lqm laboratory. Introduction to peripheral blood smear examination clinical. Before staining and observing a microbe under a microscope, a smear must be prepared. Preparation of thick blood films and giemsa staining. Making and staining a bloo d smear a wellmade blood smear is a beauty to behold, and likely to yield interesting and significant information for a research project. A properly prepared blood smear is essential to accurate assessment of cellular morphology. Films made on coverglasses have negligible advantages. 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 giemsa blood smear test is a test to detect blood parasites. 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.
The nucleus, which is acidic in character, has an affinity for basic stains and the. The bone marrow stroma has fibroblasts, macrophages, adipocytes, osteoblasts, osteoclasts, endothelial cells. Staining of blood film after preparation, the slide is stained to distinguish the cells from each other. This sample is sent to the laboratory for testing within two hours of collection. 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. An introduction to haematology and blood smear preparation. Wait until the thin and thick films are completely dry before staining. An extensive study was undertaken to compare five methods of blood smear preparation. As soon as the drop of blood is placed on the glass slide, the smear should be made without delay.
Clean, highquality glass microscope slides are essential to blood smear preparation. Bacteria have nearly the same refractive index as water, therefore, when they are observed under a microscope they are opaque or nearly invisible to the naked eye. Blood smears are needed for microscopic examination of the blood. Giemsa staining technique principle, preparation, procedure. Blood filled capillary tube or purple top with diffsafe dispenser. Preparation of a peripheral blood film this protocol describes how to prepare a slide film of peripheral blood for microscopic examination.
A wellmade blood smear is a beauty to behold, and likely to yield interesting and. Smears made from edtaanticoagulated blood should be made. Start a free trial of quizlet plus by thanksgiving lock in 50% off all year try it free. The purpose of preparation of blood smear blood film is to study the morphology of rbcs, differential leukocyte count and reticulocyte count. Preparation and staining methods for blood and bone marrow.
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