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