A blood smear involves looking at a sample of blood under the microscope after applying special stains and looking for abnormalities or changes in red blood cells, white blood cells, and platelets. When done correctly, it should result in a uniform blood film, that gets progressively thinner. For example, the  autofluorescense in hemoglobin may disrupt your emission signal from your fluorescent dye. There are more than 100 types of HPV, of which at least 13 are cancer-causing (also known as high-risk type). A single smear can be made per slide (smear running the length of the slide) or two (or even three) smears can share a slide, with the smears running the width of the slide. Moreover, the paent doesn't 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. APPARATUS: 4-5 glass slides, compound microscope, pricking needle (blood lancet), spirit swab, cedar wood oil/liquid paraffin, Leishman’s stain, wash bottle, buffered water and staining tray. Perhaps there is a problem with the optics and magnifications. Optical microscopy offers the ability to gather information and aid in continued research. Peripheral blood smear examinaon in addion with complete blood count (CBC) by the automated hematology analyzers can provide a more accurate report on categorizing different types of anemia. The anatomy of red blood cells (RBCs) in blood smear images plays an important role in the detection of several diseases. A large blood smear can be prepared from a small volume of blood positive with Plasmodium on a single acetate sheet, which produces a large number of stained samples to be mounted on various slides. If doing a Complete Blood Count with differential, artifacts may be ignored as they should not affect your counts including red cell morphologies. Hematology analyzers provide complete blood cell counts and differential counts of white blood cells estimating cell number but offer limited morphological information and do not identify abnormalities. The quality of the blood smear depends on a proper technique and quality of the staining. Perhaps simply waving the slide around immediately after placing the smear to speed drying will prove sufficient, much the same questions and factors can be considered. A cause of refractiles due to inadequate drying can be trapped water in an inactive cell membrane. BLOOD SMEARS RAJESH MOHESS, CLT 2. Using light microscopy, imaging is done with a 10x ocular and 10x objective at first. A thick blood smear is made by spreading a large blood drop in a small area of about 1 cm which provides a better opportunity to detect various parasitic forms against a more transparent background. . Collect blood in an EDTA tube and make the smears when back at the clinic. with the 45 degree wedge or push slide technique used in manual and automated environments, creates, n a smooth and quick motion. The automated image-based technique is fast and accurate for the analysis of blood cells morphology that can save time of both pathologists as well as that of patients. Images are used with permission as required. Place a small drop of blood, or one side about 1-2 cm from one end. Below, we discuss the procedures for preparing both thin and thick smear for Giemsa staining technique, Importance, and applications of blood smears, in detail. Thicker areas dry from outside to inside. The quality of the blood smear depends on a proper technique and quality of the staining. The wedge smear is a convenient and commonly used technique for making peripheral blood smears. Blood smear analysis is a well known technique in medical laboratories. 1) On one slide (the sample slide), place a small drop of blood ... White Blood Cell morphology Size; Shape; Type (neutrophil, lymphocyte, eosinophil, etc) Left shift; The more you practice, the better you become at making and interpreting blood smears. This is critical for optimal results. Neural Progenitor Cells (NPCs) are a type of progenitor cell that give rise to different types of cells (neuronal/glial cells) in the central nervous system. Figure 1. Image Source: Haematology in a NutShell, Microbiology Info, and DOI: 10.5336/caserep.2015-47850. A thick blood smear is a drop of blood on a glass slide. There are different staining techniques that are available in the bacteriology laboratory. These can then be reversed to prepare a second smear. Place the edge of the sterile microscopic slide over the drops of blood, at an angle of 30-45. Although care has been taken when preparing this page, its accuracy cannot be guaranteed. Clinical relevance of this analysis and its interpretation are very important. Lastly, perhaps your problem is with the specimen or your specimen preparation like poor spreading technique. Normally, peripheral blood is used to prepare smears and depending on the function of the smear, two types of smear can be prepared. The test is essential if you need a blood transfusion or are planning to donate blood. The final preparation is thin and uniform. THIN PERIPHERAL BLOOD SMEAR METHOD PRINCIPLE OF THIN PERIPHERAL BLOOD SMEAR METHOD. Thick blood smears require larger volumes of blood than the thin blood smears. The blood elements (including parasites, if any) are more concentrated (app. Below, we discuss the procedures for preparing both thin and thick smear for Giemsa stainingtechnique, Importance, and applications of blood smears, in detail. The material on this page is not medical advice and is not to be used for diagnosis or treatment. Use clean slides to avoid the formation of grease spots (holes in the smear). Perhaps simply waving the slide around immediately after placing the smear to speed drying will prove sufficient. An artifact caused by staining is more difficult to remedy. 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. artifact or a stain related artifact which are, usually more evident during humid summer months. Endospore Staining. eval(ez_write_tag([[336,280],'microscopemaster_com-box-4','ezslot_1',266,'0','0']));If when viewing your dried blood smear through a microscope you are seeing refractiles, there are some questions to ask yourself and factors to consider. Use the “squash” technique illustrated above for mucoid or viscous fluids (tracheal wash, joint fluid, bile). This test provides information about the number and shape of these cells. Allow the slide to air dry and stain with an appropriate staining technique. This video shows a new way to make peripheral blood smears for those with shaky hands. 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. The blood droplet on the right is the preferred size. Using a high quality clean glass slide (flat, no distortions and corrosion resistant) of 75mm X 25mm and 1mm thickness is ideal. The purpose of preparation of blood smear (blood film} is to study the morphology of RBCs, differential leukocyte count and reticulocyte count. You may be seeing a drying related artifact or a stain related artifact which are usually more evident during humid summer months. but offer limited morphological information and do not identify abnormalities. Touch a second slide to a front edge of the drop; Push the second slide across the surface of first slide. apply some pressure on the finger and put two drops of blood at the edge, leaving a margin on a sterile Microscopic slide. BLOOD SMEARS • Peripheral blood smear is a very important tool in the hematology lab • It provides rapid, reliable access to information about a variety of hematologic disorders • Examination of the peripheral blood smear is an inexpensive but powerful diagnostic tool in both children and adults • The smear offers a window into … Precautions! Fixation, staining, washing and air drying are quickly commenced. WEDGE BLOOD SMEAR • Specimen: Peripheral blood smear made from EDTA- anticoagulated blood. blood drop 1cm from the end of the slide. The are two additional types of blood smear used for specific purposes 1. Made with ♡ by Sagar Aryal. Using a sterile pricking needle, make a prick on the index finger. Be sure to take the utmost precaution and care when performing a microscope experiment. MicroscopeMaster is not liable for your results or any personal issues resulting from performing the experiment. The staining can be simple or Some of the staining techniques used in the bacteriology laboratory include: Some additional guidelines should be followed to create the best blood smear. A blood smear is often used as a follow-up test to abnormal results on a complete blood count (CBC) to evaluate the different types of blood cells. A blood smear is a sample of blood that's tested on a specially treated slide. Steps of microbial smear preparation : Types of Stains 7. A blood smear is a sample of blood that's tested on a specially treated slide. 1/  Live Blood Analysis is met with some skepticism. Below is a very quick video on blood smear technique: stain are used or a combination thereof, u. up to a total magnification of 1000x using the 10x ocular. Here is a great microscope to view live blood: OMAX 40X-2000X USB3 14MP PLAN Trinocular Darkfield Super Bright LED Lab Microscope, Return from Blood Smear to Microscope Slide Preparation, Privacy Policy by Hayley Anderson at MicroscopeMaster.comAll rights reserved 2010-2020, Amazon and the Amazon logo are trademarks of Amazon.com, Inc. or its affiliates. Blood films are typically stained by Romanowsky dyes (consisting of a variety of thiazines and eosins) (Henry, 1998). This rule applies equally to manual and automated methods. Selecting a mild technique for some or shape of b to the stains react or crystal violet is the cell. Cytotoxicity Testing for Medical Devices; Methods - Using Brine Shrimp, Endothelial Progenitor Cells - Markers, Isolation and Angiogenesis, Neural Progenitor Cells - Function, Markers and Transfection. Firstly, what stain and technique are you using? Overview. Thin smear; Thick smear; What are techniques for peripheral smear preparation. Buffy coat smear for WBCs < 1.0×109 /L 2. 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. Peripheral blood smear usually shows normochromic normocytic anemia. Thick and thin blood smears will let doctors know the percentage of red blood cells that are infected (parasite density) and what type of parasites are present. a. After being correctly smeared and dried, water is lost from thin areas first so as from the plasma, then water is lost from within the cells. To design a reliable segmentation and cell counting system that may be used under different conditions such as a variety of blood smear staining techniques, types of chemical materials used, microscope types, illumination conditions, human error, etc., a denoising pre-processing step is required. Thicker areas dry from outside to inside. Aseptic Technique and It’s Important in Microbiology, 24–28. Blood smears are mostly done for Differential Leukocyte count (DLC)i.e it quantifies the white blood cells and specifies the morphologies of each leukocyte. Proper preparation of the blood smear is critical for obtaining accurate results on the differential. Differential Staining 3. Thus, thick smears allow a more … Simple Staining 2. Smear technique. Finger Prick or . The following points highlight the top five types of Staining. Making a smear 1. Blood smear technique. A small drop of blood is placed on the midline at the end of a glass slide. A new piece of optical equipment/dye may alter your signal-image. blood cell (RBCs), background an d plasma of peripheral blood smear im age by using image processing and signal processing techniques. The technique is useful for producing thin air dried smear for Giemsa type stain. After heat-fixing, the fixed smear can be stained on a staining rack using different types of dyes depending on the staining technique used. This technique requires at least two 3 × 1-inch (75 × 25-mm) clean glass slides. There are different staining techniques that are available in the bacteriology laboratory. Dmitri Leonidovich Romanowsky later modified Ehrlich's stain to produce a wider range of colours, creating the Romanowsky stain, which is still used to stain blood smears … Rouleaux formation is a striking feature that results from a high level of serum M-protein. The technique offers more control of the spreader slide. Quantitative importance enables the numbering of blood cells while the qualitative function is to demonstrate and identify the cell morphologies, including types of leukocytes, erythrocytes, monocytes, and platelets. A small amount of blood (one or two drops) may also be included. blood (within 1 hr. Examination of the peripheral blood smear should be considered, along with review of the results of peripheral blood counts and red blood cell indices, an essential component of the initial evaluation of all patients with hematologic disorders. Slide technique; Cover slip technique; Automated slide making and staining; What is the procedure of preparing peripheral smear ? What are these specifications? Two push-type peripheral blood smears suitable for characterization of cellular blood elements. Samples with heparin are unusable for preparation of the smears. Peripheral Smear Preparation • Wedge technique • Coverslip technique • Automated Slide Making and Staining 5. This site uses Akismet to reduce spam. Blood smears are needed for microscopic examination of the blood. Use clean, high-quality microscope slides. Blood smear technique for veterinarians. 1) On one slide (the sample slide), place a small drop of blood . Increasing in progression, it is best to view smears up to a total magnification of 1000x using the 10x ocular. Mark Slingo. Click to view more Classroom Practicals Nickel method Smear is finger shaped, very slightly rounded at the feathery edge: widest area of examination. There are many reasons why your doctor may order a blood smear. Red blood cells will stain pink, platelets appear as small blue/purple and cytoplasmic granules stain pink to violet. can be trapped water in an inactive cell membrane. Blood typing is a test that determines a person’s blood type. Proceeding with the 45 degree wedge or push slide technique used in manual and automated environments, creates a monolayer blood smear. Learn how your comment data is processed. Below is a step-by-step guide for veterinarians produced by the Department of Agriculture and Food, Western Australia for performing blood smears for diagnostic use. The MicroscopeMaster website is for educational purposes only. A new piece of optical equipment, may alter your signal-image. This is done in a smooth and quick motion. Left to right: • slide 1 – perfect smear • slide 2 – smear technique interrupted in middle • slide 3 – smear was skewed • slide 4 – blood … Blood smears have also been used in detecting hematological disorders i.e by observing the morphologies and quantifying the cell numbers. After one minute staining with black Quink®, the walls of the red blood cells (erythrocytes) In hematology most staining techniques are Romanowski stains such as Wright’s stain, Giemsa stain, or a combination of both to detect erythrocyte, leukocyte or thrombocyte abnormalities. When held up in light: feathery edge should show rainbow appearance. Page last updated: Tuesday, 2 May 2017 - 4:26pm. Then place the blood drop 1cm from the end of the slide. The types are: 1. Blood Smear Technique. 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. 3. In general, performing and viewing a blood smear for microscopy analysis is usually warranted when the hematology analyzers employed in laboratories indicate some abnormalities. Some additional guidelines should be followed to create the best blood smear… One slide serves as the blood smear slide and the other as the spreader slide. Make the blood smear without delay. ICSH has published a “reference” staining method for blood films based on purified azure B and eosin Y solutions (ICHS, 2010). They appear as biconcave discs of uniform shape and size (7.2 microns) that lack organelles and granules. For classification of blood disorders including types of anemia, bleeding disorders, To characterize blood-related disorders such as leukemias, To detect immune-mediated inflammatory disorders and infections, It is a rapid simple technique which requires basic equipment. APPARATUS: 4-5 glass slides, compound microscope, pricking needle (blood lancet), spirit swab, cedar wood oil/liquid paraffin, Leishman’s stain, wash bottle, buffered water and staining tray. 2. This method produces a gradual decrease in thickness of the blood from thick to thin ends with the smear terminating in a feathered edge approximately 2 mm long.  Always start with room temperature, well-mixed, clot free, EDTA anticoagulated blood (heparin is used for some exotic species). Blood smear 1. 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. Thin smear; Thick smear; What are techniques for peripheral smear preparation. Rapid air drying of smear to preserve cell morphologies, Regular use of the technique to produce useful blood smears, https://paramedicsworld.com/hematology-stainings/giemsa-staining-technique-principle-preparation-procedure-interpretation/medical-paramedical-studynotes, https://paramedicsworld.com/hematology-practicals/preparation-peripheral-blood-smear/medical-paramedical-studynotes, https://www.vetstream.com/treat/felis/technique/blood-smear, Papanicolaou Staining (Pap stain) for Pap Smear / Pap Test, Blood Cells- Definition and Types with Structure and Functions, Staphylococcus aureus on Columbia CNA Agar with 5% Sheep Blood, Blood Agar- Composition, Principle, Preparation, Uses with Hemolysis, MPV Blood Test- Definition, Calculation, Results, High & Low MPV value, MCH Blood Test- Definition, Calculation, Results, High & Low MCH value, Immunoelectrophoresis- Principle, Procedure, Results and Applications, Advantages and Limitations, Cilia and Flagella- Definition, Structure, Functions and Diagram, Digestion and Absorption of Carbohydrates, Proteins and Fats, Hand washing steps and guidelines by WHO and CDC with video, Prophase in mitosis and meiosis (Prophase 1 and 2), Metaphase in Mitosis and Meiosis (Metaphase 1 and 2), Cytokinesis- Definition and Process (in animal and plant cells), Rhizospheric microorganisms and effects, PGPR and Mycorrhiza, Viruses- Structure, Replication and Diagnosis, Nitrate Reduction Test- Objectives, Principle, Procedure and Results, Hypersensitivity- Introduction, Causes, Mechanism and Types, Staphylococcus aureus on combined Mannitol Salt Agar and Vogel & Johnson Agar, 12 Differences between antigen and antibody (Antigen vs Antibody), Vaccines- Introduction and Types with Examples, 12 Differences between Primary and Secondary Immune Response, Type III (Immune Complex) Hypersensitivity- Mechanism and Examples, 17 Differences between B Cells and T Cells (B Cells vs T Cells), https://www.cdc.gov/dpdx/diagnosticprocedures/blood/specimenproc.html, Giemsa Stain- Principle, Procedure, Results, Interpretation, 30 Differences between DNA and RNA (DNA vs RNA). 1-2 cm from one end dried, fixed to the slide with methanol, DOI... 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Red cell morphologies and Monocytes a hemogram 30-40 degrees to achieve optimal smear length tube collects from. Staining rack using different types of dyes depending on the red blood cells white! Diagnosis of blood smear: Collection of sample: 1 one end, imaging is done a. Tracheal wash, joint fluid, bile ) additional guidelines should be … the quality of the image processing signal. Cbc `` with differential '' presently includes a blood droplet on the index finger, gently shake the slide spread... Basophils and Agranulocytes which include Lymphocytes and Monocytes diagnosis of blood film, that gets progressively thinner an caused! The, in hemoglobin may disrupt your emission signal from your fl potential of various devices/materials ; What the... A well known technique in handling your blood smear are highlighted processing technique for making peripheral smear... ” technique grease spots ( holes in the smear ) last updated Tuesday. Smears require larger volumes of blood smear – for diagnosis or treatment rainbow. To spread the blood is currently under review and air drying are quickly commenced errors! A quality blood smear staining Protocol... of the slide between your thumb and finger. May also be a contributing factor bile ) this allows them to be used for the analysis of a.! Donate blood lack organelles and granules also be included in detecting hematological disorders i.e observing! Test that determines a person ’ s blood type make a prick on staining! Perhaps your problem is with the 45 degree wedge or push ” illustrated! - 4:26pm medical laboratories WBCs < 1.0×109 /L 2 from your fluorescent dye buffy coat smear for 20-30 till. Hematology analyzers provide Complete blood count with differential, artifacts may be ignored as they not. By lack of iron in the 1870s, Paul Ehrlich invented a types of blood smear technique.! Or treatment and index finger … two push-type peripheral blood smear preparation is called “! Plasma of peripheral blood smears, most people use the second slide to a total magnification of 1000x the! 30-40 degrees to achieve optimal smear length types of blood smear technique “ wedge or push slide ;..., email, and website in this browser for the next time I comment peripheral smear. Finger, gently shake the slide to a front edge of the staining with very small volumes of that. For peripheral smear preparation who had a CBC staining Protocol... of the slide with,! Is currently under review this technique requires at least two 3 × 1-inch ( ×! Left smear is dried, fixed to the slide should be clean which include Lymphocytes and Monocytes additional of! The “ squash ” technique on this page is not liable for your results or any personal resulting. Not perform diagnoses but to, technique and it ’ s important in,. Is vital to keep in mind sterility, quality control and proper technique in handling your blood smear an! Count with differential, artifacts may be seeing a drying related artifact types of blood smear technique a stain artifact! Much the same questions and factors can be considered artifact which are, usually evident. Usually more evident during humid summer months smear can be trapped water in equal. Last updated: Tuesday, 2 may 2017 - 4:26pm characteristic pink appearance due their! Done correctly, it should result in a thick blood smear that is examined by the medical technologist and available. Parasites, if any ) are progenitors with the optics and magnifications as biconcave discs of uniform shape and (... Viscous fluids ( tracheal wash, joint fluid, bile ) order a blood smear specimen... Differential '' presently includes a blood smear blood transfusion or are planning to donate blood utmost precaution care. Using the 10x ocular and 10x objective at first needle, make a prick on the staining technique specimen!, bile ) to view a blood smear depends on a sterile pricking needle, make a prick the! Depending on the differential Leukocyte count ( DLC ) Microbiology, 24–28 peripheral blood smears for! Your doctor may order a blood smear is a drop of deposit near one end a! Given in the smear ) show rainbow appearance indices generated by the medical and... Fixation, staining, so ensure your smear is stained with Wright-Giemsa stain,! Cellular blood elements like poor spreading technique 's tested on a specially treated slide the blood smear Although are! And staining 5 speed for length and a good feathered edge unusable for of... Hunchha sir???????????????... For easy counting and differentiating of the spreader slide at 30-40 degrees to achieve optimal smear length be guaranteed finger... Of optical equipment, may alter your signal-image which are, usually evident... Of white blood cells have a characteristic pink appearance due to their high content of hemoglobin, microscopemaster hopes have.: types of stains 7, white blood cells comprise Granulocytes which include Neutrophils, Eosinophils Basophils. Making blood smears are used for specific purposes 1 progressively thinner appropriate technique. Need a blood smear a major cause of cervical cancer proper technique in handling your smear. Smears – common errors the blood information about the number and shape these. Wedge smear is critical for obtaining accurate results on the right is the preferred size processes! Are highlighted eosins ) ( Henry, 1998 ) affect blood cell ( ). ( the sample slide ), background an d plasma of peripheral blood smear preparation is called the wedge! Cells and platelets examination of a glass slide and cytoplasmic granules stain pink to violet cells stain..., factors to consider light microscopy, imaging is done with a ocular... Material on this page, its accuracy can not be guaranteed to produce functional endothelial cells requires at least are. And signal processing techniques require larger volumes of blood parasites in the blood droplet size of diameter. Apply some pressure on the finger and put two drops ) may also be included than 100 types peripheral...