PRINCIPLE: Blood smear is prepared, stained with Leishman's stain and cells are identified under oil immersion lens BLOOD SMEAR BASICS JENNIFER 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 point-of-care hematology analyzers, examination of a well prepared, well stained blood smear remains the cornerstone of veterinar Procedure: Examination of the Peripheral Blood Smear (How to Perform) I. PRINCIPLE The examination of the peripheral blood smear is an important basic hematological procedure. Many hematological diagnoses depend upon this procedure and often a definitive diagnosis can be established from the careful examination of the blood film Prepare a film of blood or bone marrow on a microscopic slide and allow to air dry. Place the air-dried smear on the slide staining rack, smear side facing upwards. Cover the blood film with undiluted staining solution. The undiluted stain fixes and partially stains the smear
Principle The Thin Blood smear is prepared by making a drop of well-mixed venous blood, 2mm in diameter at the center of a sterilized microscopic glass slide. Some borders are left around the smear for easy counting and differentiating of the cells PREPARATION OF A BLOOD SMEAR Principle: • A well-stained peripheral smear will show the red cell background as red orange. • White cells will appear with blue purple nuclei with red purple granules throughout the cytoplasm. • A well made, well distributed peripheral smear will have a counting area at the thin portion of the wedge smear. The blood smear test is a simple procedure in which your health care provider draws a blood sample from the vein in your arm. The blood sample is sent to the lab where a drop of blood is spread thinly onto a glass slide and it is then treated with a special strain. This procedure is known as a blood film Giemsa stain procedure for thin smear (Blood) For thin smear, prepare a 1:20 ratio of giemsa stain by mixing 2 ml of stock solution of Giemsa stain to 40 ml of phosphate buffer solution. You may use Distilled water instead of buffer
Principle of Smear Preparation. - small drop of blood is placed near the frosted end of a clean glass slide. - second slide is used as a spreader and blood is streaked. - air dry then stained. Specimen. - EDTA anticoagulated blood is preferred (lavender top) - can also make from finger stick directly on a slide. Notes The method for staining, concentration and timing of stain used varies according to the purpose, for example, thin blood smears use 1:20 dilution of stock whereas for thick blood smear 1:50 dilution is used. For Thin blood smear . Fix air-dried film in absolute methanol by dipping the film briefly (two dips) in a Coplin jar containing absolute.
Blood smear staining 1. Preparation of blood smear with different staining method Dr. Ankur Patel Ankurvety001@gmail.com 2. Blood film ???? • 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 Peripheral Blood Smear Preparation. A reproducible blood smear review requires every peripheral smear be prepared for consistent cellular distribution and proper clarity. Well-made peripheral smears can be prepared by starting with only a drop of blood at one end of a clean glass slide. The drop is smeared lightly and quickly with a wedge. . The three main blood cells that the test focuses on are: The test provides information on the number and shape of these. A blood smear is a sample of blood that's tested on a specially treated slide. For a blood smear test, a laboratory professional examines the slide under a microscope and looks at the size, shape, and number of different types of blood cells. These include: Red blood cells, which carry oxygen from your lungs to the rest of your body Millones de Productos que Comprar! Envío Gratis en Productos Participantes
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. GENERAL PRINCIPLES OF COMPONENT PREPARATION. The Whole blood is collected as 350 ml or 450 ml in double/triple/quadruple or penta bags with CPDA-1 or additive solution. After blood collection, components should be separated within 5 - 8 hours. Component room should be a separate sanitised room
A. Proper Preparation of a Peripheral Blood Smear. Objectives. At the completion of this laboratory, you will be able to: 1. State the appropriate sample used for preparing a peripheral blood smear. 2. Describe the appearance of a well prepared blood smear. 3. Demonstrate the appropriate technique for preparing a peripheral blood smear. 4 Procedure. - Placing a drop of blood from a mixed sample on a clean glass slide. - Spreader slide: using second clean glass slide at a 30-40-degree angle. - Control the thickness of the smear by changing the angle of the spreader slide. - Let the blood film to air-dry completely before staining
Delay in preparation of blood smear may allow for the degeneration of the cellular elements of blood and may result in a pseudo-thrombocytopenia (falsely reduced platelet count) due to formation of platelet aggregates. 2. Slide preparation is done by trained personnel preferably a medical laboratory technologist, who can ensure quality slides. Smear examination. Examination of a blood smear is an integral part of a hemogram. Blood smear analysis allows quantitation of the different types of leukocytes (called the differential count), estimation of the platelet count, and detection of morphologic abnormalities that may be indicators of pathophysiologic processes
PREPARATION OF BLOOD SMEAR (WEDGE METHOD) (1) A small drop of blood (2-3 mm in diameter) is placed in the center line about 1 cm away from one end of a glass slide (typical size of slide is 75 × 25 mm; thickness about 1mm) with a wooden stick or glass capillary. Slide should be clean, dry, and grease-free A single drop of blood is placed on the surface of a clean and grease-free microscope slide at a distance of 2 cm from one end. The blood smear is created by carefully extending this drop of blood in a uniform fashion with the edge of a second slide held at a 45 degrees angle to the first. Once prepared, the blood smear slide is dried by gently. Staining the Blood Film After a blood film has been prepared, the next step is promptly staining the blood smear; if it cannot be stained within a few hours, it should be fixed by immersion in absolute methyl alcohol for 1 or 2 seconds and air dried. The stain most often used for the examination of blood films is Wright's stain or a variation, Wright-Giemsa stain. Romanowski stains contain. Principle Whole blood is diluted with a 1% ammonium oxalate solution. The isotonic balance of the Prepare two leukocyte/platelet Unopettes as follows: a. peripheral blood smear estimate should be repeated. 6. In acute leukemia, there is an increase of blast cells in the peripheral blood. Ofte
I. Principle The combination thick-thin blood film provides both options on one glass slide and the slide can be stained as either a thick or thin blood film. If fixed prior to staining, then the smear will be read as a thin blood film; if RBCs are lysed during staining, the preparation will be read as a thick blood film (parasites contact with the blood and glass, rotate (do not roll) the stick in a circular motion while moving the stick down the glass slide to the opposite end. 4. The appearance of the blood smear should be alternate thick and thin areas of blood that cover the entire slide. 5 A peripheral blood smear can be prepared from A. EDTA-anticoagulated blood within 1 hour of collection B. free-flowing capillary blood C. citrated whole blood D. both A and B: 21. Identify the characteristic(s) of a good peripheral blood smear. A. It progresses from thick at the point of origin to thin. B. It has a blunt feathered termination. C
The wedge smear is a convenient and commonly used technique for making peripheral blood smears. This technique requires at least two 3 × 1-inch (75 × 25-mm) clean glass slides. High-quality, beveled-edge microscope slides are recommended. One slide serves as the blood smear slide and the other as the spreader slide Smear slides require two or more flat, plain slides, cover slips, pipette and tissue paper: Pipe a liquid sample such as blood or slime onto a slide. Using the edge of the second slide, slowly smear the sample creating a thin, even coating. Put a cover slip over the sample, careful not to trap air bubbles. Remove excess liquid Result Interpretation of vaginal smear or vaginal wet mount. Bacteria: The organisms which have size of this range 0.2 - 2.0 × 1 - 10 µm and shape of normally round or rod. Trichomonas vaginalis: A moving object usually the size of 10 μm in length and 7 μm in width. Yeast cells: Oval or round in shape having size of usually 3-5µm and. State the principle of the occult blood including the chemical reaction involved. 2. The confirmatory test for diagnosis of blood parasites is made through the examination of fresh blood and preparation of blood smears which are then examined for the presence of the parasites. Examinations of blood smears are very time consuming..
The thick blood film permits the examination of a large amount of blood for the presence of parasites. Since the smear is not fixed with methanol, the red blood cells (RBCs) lyse, permitting better visualization of the organisms. The thin film allows for the observation of RBC morphology, inclusions, and intracellular and extracellular parasites Principle And Interpretation Leishman stain, is used in microscopy for staining blood smears. It provides excellent stain quality. It is generally used to Specimen Collection and Handling For clinical samples follow appropriate techniques for handling specimens as per established guidelines( 1,2) Thick blood smear: The standard method for diagnosing active infection is the identification of microfilariae in a blood smear (multiple thin and thick blood smear) by microscopic examination. Blood collection should be done at night to coincide with the appearance of the microfilariae, and a thick smear should be made and stained with Giemsa. The blood specimen to be used for differential leucocyte count is capillary blood or EDIA anti-coagulated blood. ADVERTISEMENTS: There are three major steps involved in differential cell count: 1. Preparation of blood smears. 2. Staining of blood. 3. Staining of smear Principles of the preparation of Liquid Based Cytology (LBC) slides. A sample of cells is collected from the cervix in the normal way using a spatula or broom sampling device. The sample is transferred into a container of preservative/ transport medium. The cell are dispersed in the fluid. An aliquot of the suspension is selected for processing
. 1 a) Poor quality manual peripheral blood smear and b) Good quality automated smear b) The staining of the blood smear The peripheral blood smear needs to be stained so that the cytoplasmic and nuclear details of the various cell types are accentuated. Romanowsky stains or derivations thereof such as Wright, Wright-Giemsa and May-Grünwald. For best result blood should be less than 6 hours old, although successful staining has been achieve on specimens refrigerated for up to 2 weeks The smears should be fixed within 2 hours of preparation. Principle Blood smears are fixed with ethyl alcohol and then incubated in a citric acid-buffer solution In an acid medium (pH 3.2 to 3.3.
Importantly, v iewing blood smears under the microscope needs to be done shortly after blood collection employing sterile technique (**wearing gloves) from a disinfected site (wiping off 1st drop of blood). Using a high quality clean glass slide (flat, no distortions and corrosion resistant) of 75mm X 25mm and 1mm thickness is ideal. T hen plac e the blood drop 1cm from the end of the slide Field stain is a histological method for staining of blood smears. It is used for staining thick blood films in order to discover malarial parasites. Field's stain consists of two parts - Field's stain A is methylene blue and Azure 1 dissolved in phosphate buffer solution; Field's stain B is Eosin Y in buffer solution PREPARATION: Giemsa and May-Grünwald solutions are supplied ready to use, although the Giemsa solution may be diluted 1:20 before use in either deionized water or in phosphate buffer solutions. Prepare Working Phosphate Buffer by diluting contents of one vial Phosphate Buffer pH 7.2 at 25°C to 3.8 liters or 1 gallon with water
A blood smear is a blood test used to look for abnormalities in blood cells. The three main blood cells that the test focuses on are: red cells, which carry oxygen throughout your body. white cells, which help your body fight infections and other inflammatory diseases Wright's Stain : Preparation, Principle, Procedure and Results July 25, 2019 Dhurba Giri 4. Wright's stain is a type of Romanowsky stain, which is commonly used in hematology laboratory for the routine staining of peripheral blood smears. It is also used for staining bone marrow aspirates, urine samples an
Peripheral blood smear is an important part of the diagnostic work-up for patients. It is a basic and simple test to perform and available in most laboratories. However, it relies heavily on the skill and ability of the reader to recognize the normal and abnormal forms of blood cells PRINCIPLE: A blood film appropriately prepared and stained is seen under the microscope in oil immersion. Different WBCs are seen which are counted in percentage. The total 100 leukocytes are studied, identified and recorded from a blood smear. PROCEDURE: Prepare, stain and examine a blood smear under oil immersion lens Nonmoving organisms may be difficult to identify within the stained smear. 1,2. Step by Step: Stained Fecal Smear 1,2. Mix a small amount of feces with saline on a microscope slide, creating a thin layer. Using a cover slip, move any fecal material out of the way (to the side) before placing the cover slip on the smear
PRINCIPLE Preparation of thick and thin blood smears, appropriate staining procedure and detection and identification of hemo-parasites are crucial to clinical diagnosis of many parasitic diseases. These include species of malaria, trypanosomes, babesias and microfilariae of filarial nematodes A blood film—or peripheral blood smear—is 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. 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 filariasi BLOOD FILM EXAMINATION • Hypochromic - central pale area becomes larger, decrease hemoglobin content, MCH and MCHC are decreased (e.g., IDA, • Preparing a smear thalassemia) → Wedge method (2-slide method) - convenient, easy • Hyperchromic - less or no central pallor (e.g., HS) handling • Anisochromia - dimorphic anemia. Two drops of blood (about 20μl each) were collected on a clean microscope slide. One drop was used to prepare a thick smear and the other was used to prepare a thin smear according to Cheesbrough (2005). Finally the slides were labelled with participant code and packed into slide porter after being air dried (Warhurst and Williams, 1996) Thick and thin blood smear preparation pdf Background: Due to the initial inexperience of students, slides are frequently broken and blood banners are damaged in the microscope training, which leads to the need for their constant replacement. principle of preparation of thick and thin blood smear. reasons for making routine thin blood film.
The Gram staining technique is the most important and widely used microbiological differential staining technique. It was developed by Dr. Christian Gram in 1884, and categorizes bacteria according to their Gram character (Gram positive or Gram negative).. In addition this stain also allows determination of cell morphology, size, and arrangement.It is typically the first differential test run. A blood smear preparation device includes a base; a carrying table for carrying a microscope slide thereon and being supported on the base; a lifting mechanism mounted to the base; a retaining stand suspended from an output end of the lifting mechanism; a spreader holder rotatably suspended from the retaining stand about a second rotating shaft and positioned above the carrying table; a. The smear is washed again, blot dried and examined under microscope (Figure 3.7). Principle of Gram's Staining. The exact mechanism of action of this staining technique is not clearly understood. However, the most acceptable explanations are associated with the structure and composition of the cell wall
The modified Knott's method is used for the concentration and identification of microfilariae, specifically the heartworm Dirofilaria immitis. It must be differentiated from the non-pathogenic microfilaria of Dipetalonema reconditum (Dipet for short). A direct blood smear can be done at the same time Smear preparation 2. Staining methods a. Ziehl-Neelsen staining b. Kinyoun's staining c. Two-step staining radiometric technique was widely used for blood culture using the BACTEC 460 instrument. 2. Principle of detection and drug susceptibility testin preparation of a direct smear, select the cheesy necrotic, and blood tinged particles in the specimen, because, they are most likely to produce a positive direct smear result. Positive results help to confirm clinical suspicions. The direct smear is not as sensitive as concentrated smear, so a direct smear shoul
Isolation of RNA from Blood - Principle, Protocol , Functions of Reagents. RNA extraction is simply the process of extraction of purified RNA from the source. The source can be anything blood sample bacterial cell animal cell or plant cell. Extraction of RNA is a comparatively sensitive process because RNA is not as stable as DNA as it is. Preparation of blood films on slides. Blood films should be made on clean glass slides. Films made on coverglasses have negligible advantages and are unsuitable for modern laboratory practice. Films may be spread by hand or by means of an automated slide spreader, the latter being either a stand-alone instrument or a component of an automated. • Blood smear prepared for you by an experienced technician • Smear evaluation performed by a technician on every specimen; provides valuable information about red blood cell and white blood cell morphology and blood parasites • Automatic pathologist review performed when results are markedly abnormal based o Capillary Puncture Principles • Blood smear preparation: - Diagnosed by presence of organism in peripheral blood smear - A very large drop of blood is placed in center of glass slide - Drop is spread with corner of another slide or cover slip until it is the size of a dim
. It looks at the appearance, number, and shape of your red and white blood cells and platelets to see whether they are normal. A blood smear can also detect parasites in your blood. It is now more common to have blood analyzed by a computer. But blood smears may still be routinely done to look for certain. 4. BLOOD FILMS Method of preparation Collection of specimen Special smear preparation Buffy coat smears 5. ROMANOWSKY STAINS Principle Types of stains Composition and Preparation Method of staining 6. RED CELLS INDICES MCV MCH MCHC,RDW,Red cell histogram. 7. STORAGE OF BLOOD SAMPLES 8 Principle. Giemsa stain is a Romanowsky stain that is widely used in parasitology to stain malaria and other blood parasites. In microbiology, the Giemsa technique can be used to stain Chlamydia trachomatis inclusion bodies, Borrelia species. Giemsa stain can also be used as substitute stain when Wayson's stain is not available, to stain Yersinia pestis Staining of Blood Smear Principle Acidic dye unites with the basic components from MED 402 at University of Michiga
Hematology blood smear, hematology, leishman stain, leishman Principle, procedure, results, advantages & disadvantages kinase alkaline phosphatase clinical enzyme constituents of Leishman stain leishman stain preparation procedure urine how to prepare leishman stain leishman stain protocol stain haematology what is prothrombin time test. the manual smear that starts with uniform smear making with feathered edge. A quicker push at a higher angle results in a thicker smear. A larger drop of blood results in a longer smear. Fig-1: Manually made smear - note the uneven spread and staining with vacuoles Fig-2: Automated analyser- Even smearing and staining CONCLUSIO . The Blood Film Master Advanced is a bundle of two top products: our hemoslider and the automated RAL Stainer, which uses the excellent MCDh reagents. Blood film preparation and staining are simple procedures that can fail or be less effective than they could be zdescribe the principle of staining zexplain red cell abnormalities seen on the smear and their interpretation 9.2 STAINING OF PERIPHERAL SMEAR Peripheral blood smears can be prepared from fresh blood to which no anticoagulant is added or from EDTA blood. Clean glass slides should be used for making blood films
SMEAR PREPARATION. The preparation of a smear is required for many laboratory procedures, A blood smear is a blood test used to look for abnormalities in blood cells. The basic principle of Gram staining is the properties of certain bacteria cell walls to retain the crystal violet dye The peripheral blood smear provides complete data on the species, the stages, and the density of parasitemia. Procedure for Peripheral Smear for Malaria Test. MP Smear is done using Smear method on a Blood sample. The microscopic tests comprise staining and direct visualization of the parasite under the microscope When observed in stained blood films, echinocytosis is usually an artifact that results from excess EDTA, improper smear preparation, or prolonged sample storage before blood film preparation. Echinocytes form when the surface area of the outer lipid monolayer increases relative to the inner monolayer Prepare thin smear of blood sample and air dry. 2. Fix smears for 3 minutes with methanol or with CytoFix fixative (Cat. No. 207030410050). 3. Stain the smear in May Grunwald stain diluted with an equal volume of distilled water for 5 minutes. 3. Put the smears without washing in 1:3 solution of Giemsa stain diluted with distilled water for 8.
This course covers the basics of normal peripheral blood cell morphology, including appearance, kinetics, and function of red blood cells, leukocytes, and platelets. It is assumed that students have a basic knowledge of the principles of cell morphology, and of preparation and staining of a Wright's stained peripheral blood smear On a stained blood smear, platelets appear as dark purple spots, about 20% the diameter of red blood cells.The smear is used to examine platelets for size, shape, qualitative number, and clumping. A healthy adult typically has 10 to 20 times more red blood cells than platelets