The PowerPoint PPT presentation: Preparation, staining and examination of blood film is the property of its rightful owner. Do you have PowerPoint slides to share? If so, share your PPT presentation slides online with PowerShow.com Procedure • Thin blood films (only) - Dip Method 1. Dip air dried blood film in undiluted stain for 15 to 30 seconds (double the staining time for bone marrow smears). 2. Decolorize the stained smears by immersion in distilled or deionized water and air dry 3. Let air dry in a vertical position World's Best PowerPoint Templates - CrystalGraphics offers more PowerPoint templates than anyone else in the world, with over 4 million to choose from. Winner of the Standing Ovation Award for Best PowerPoint Templates from Presentations Magazine. They'll give your presentations a professional, memorable appearance - the kind of sophisticated look that today's audiences expect . As an alternative for. The blood film is one of the world's most widely and frequently used tests and has undergone remarkably few changes since its introduction in the late 1800s. This article gives direction and some standardization in the preparation of blood films used for morphologic evaluation in the clinical labora
Preparation of Blood Smear: Collection of Sample: 1. Finger Prick or . 2. E.D.T.A. blood (within 1 hr. of collection) Preparation of Blood Film: The slide should be clean. Place a small drop of blood, or one side about 1-2 cm from one end. Without delay place a spreader at an angle of 45° from the slide and move it back to make contact with. Procedure. • Thin blood films (only) - Dip Method. 1. Dip air-dried blood film in the undiluted stain for 15 to 30 seconds (double the staining time for bone marrow smears). 2. Remove the color of the stained smears by immersion in distilled or deionized water and air dry. 3. Let air dry in a vertical position . Preparation of blood smears Blood smears are made from well-mixed, fresh total EDTA blood or from the resuspended buffy coat if enrichment of leukocytes is required. Pipet 2 µl blood (or buffy) in the middle of the specimen slide (untreated for MG staining, coated for IHC) ca. cm from the labeling area MODULE Staining of PBF and Interpretation of Normal and Abnormal Red Cell Morphology Hematology and Blood Bank Technique 72 HEMATOLOGY AND BLOOD BANK TECHNIQUE Notes Smears are routinely stained with Romanowsky stains with very good results. These dyes have the property of making subtle distinction in shades of staining
Procedure for Thick and Thin Blood Film Staining Technique. A thick film was made by placing a large drop of blood (about 15mm in size) on the centre of grease-free microscopic slide. Without delay, the blood was spread with a glass spreader held at a steep angle to achieve a thick smear covering an area of 15 x 15mm At least one film should be fixed for a Perls' stain on the initial bone marrow aspirate of each patient, and, if necessary, films should be fixed in the appropriate fixatives for special staining ; others should be fixed and stained with a Romanowsky stain as described later. Crushed bone marrow particles and touch preparations from trephine. The most common technique of blood smear preparation is called the wedge or push technique. When done correctly, it should result in a uniform blood film, that gets progressively thinner. A small drop of blood is placed on the midline at the end of a glass slide. Second slide (ideally narrower than smear slide, to avoid spreading the. Place the drop of blood in the center on one side of the glass slide leaving about 1 cm margins. ⇒ Place the specimen glass slide on a flat surface and hold it with the index finger and the thumb of the left hand (for Right-handed peoples). Now, Place a smooth, clean edge of the spreader slide on the specimen slide at an angle of about 30. 1)Freshly prepare and rapidly air dry blood film. 2)Cover the film with Leishman's Stain (S018S) and allow to act for 1 minute. Methanol in the stain fixes the preparation. 3)Add double the volume of distilled water to the slide and mix. 4)Allow the diluted stain to act for 10-12 minutes. 5)Wash the film with distilled water or phosphate buffer.
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. Leishman stain is a differential stain that is used to variably stain the various components of the cells and it can be used to study the adherence of pathogenic bacteria to the human cells . Al Aswad Amany S. Al Hindi. Aim of blood smear Value of blood films: Examination of thin blood films is important in the investigation and management of anaemia, infections, and other conditions which produce changes in the appearance of blood cells and differential white cell count. A blood film report can provide rapidly and at low. The blood film is one of the world's most widely and frequently used tests and has undergone remarkably few changes since its introduction in the late 1800s. This article gives direction and some standardization in the preparation of blood films used for morphologic evaluation in the clinical laboratory. Methodologies that can be labeled as being state-of-the-art and methods applicable for. 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
This section of the document describes the procedure for the preparation of blood lms from peripheral blood collected by ngerprick or venepuncture for staining and microscopy to detect, identify and quantify malaria parasites. This procedure is intended for use in malaria clinical research studies such as those assessing drug or vaccine ef cac Place 1 small drop of fresh blood on one end of the slide. Using the end of another slide at an angle of approximately 30° to the first, allow the second slide to move along the first slide to spread the blood drop into one continuous thin film. Air dry the smear before staining. Fix blood film in absolute methanol (acetone-free) for 30 seconds Preparation of a Peripheral Blood Film slide To ensure accurate and reliable diagnosis, pre-analytical variables that can affect the quality of film must be controlled. These include patient preparation and consent, blood sampling technique, transportation of the sample to the laboratory and sample preservation. Blood
Stain for 10 minutes. Prepare a 1:10 dilution of Giemsa stain with phosphate buffer pH 6.8. Mix well. After 10 minutes of May Grunwald staining, pour away the May Grunwald stain off the slides. Then pour the Giemsa mixture onto the slides and stain for another 15 minutes. After 15 minutes, pour off stain and flush the slides with running tap water 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. The smear is stained with Wright's stain and performed after the complete blood count is run 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: The test provides information on the number and shape of these. Procedure. A blood smear test is performed by first obtaining a 5 mL blood sample from the patient. The patient should be educated about the procedure before taking a sample from their vein, or.
The agreement between direct smear preparation methods from positively flagged blood culture broth with the culture smear Gram stain was statistically analyzed using kappa statistics and was found to be maximum for the blood film method (0.637), followed by drop and rest (0.570), water wash (0.496), and conventional method (0.475) and To prepare a thick blood film, a blood spot is stirred in a circular motion with the corner of the slide, taking care not make the preparation too thick, and allowed to dry without fixative. After drying, the spot is stained with diluted Giemsa (1 : 20, vol/vol) for 20 min, and washed by placing the film in buffered water for 3 min Read the latest articles of Clinics in Laboratory Medicine at ScienceDirect.com, Elsevier's leading platform of peer-reviewed scholarly literatur 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 scratches allow for improved adherence of the blood film to the slide without affecting the smear morphology. The smear can then be stained as soon as it is dry, generally within 20-30 minutes of smear preparation. Reference: Norgan AP, Arguello HE, Sloan LM, Fernholz EC, Pritt BS. Malaria Journal 2013; 12: 231. Thin smear
Specimen Preparation. Transport 5 mL whole blood (Min: 1 mL). Extended exposure to EDTA anticoagulants can result in altered parasite morphology. For best results, send 5 thin blood smears (unstained, unfixed) AND 5 thick smears (unstained, unfixed) in addition to whole blood. Thin and thick blood smears should be prepared immediately or within. peripheral blood mononuclear cells that were stimulated with or coverslip before commencing the staining procedure. Sample preparation is also intimately linked to the method of fixation, which in turn is influenced by the desired detection technique (fluorescence versus chromogenic) Heinz bodies (also referred to as Heinz-Ehrlich bodies) are inclusions within red blood cells composed of denatured hemoglobin. They are not visible with routine blood staining techniques, but can be seen with supravital staining.The presence of Heinz bodies represents damage to hemoglobin and is classically observed in G6PD deficiency, a genetic disorder that causes hemolytic anemia
Normal blood smears were stained by the standardised azure B-eosin Y Romanowsky procedure recently introduced by the ICSH, and the classical picture resulted. The effects of varying the times and temperature of staining, the composition of the solvent (buffer concentration, methanol content, & pH), the concentration of the dyes, and the mode of. A blood smear, also referred to as a peripheral smear for morphology, is an important test for evaluating blood-related problems, such as those in red blood cells, white blood cells, or platelets. It has a wide range of uses, including distinguishing viral infections from bacterial infections, evaluating anemia , looking for causes of jaundice. The SAF stool smear can also be postfixed in Schaudinn's fixative prior to staining (begin the trichrome stain protocol with the 70% alcohol rinse prior to iodine-alcohol). After drying, the smear can be placed directly into 70% alcohol (step 4) of the staining procedure (the iodine-alcohol step can be eliminated)
Blood Lab First Week This is a self propelled powerpoint study atlas of blood cells encountered in examination of the peripheral blood smear. It is a requirement of this course that you begin review of these slides with the first day of class in order to familiarize yourself with terms used in describing peripheral blood morphology 2)Treat the dried blood film with methanol for 3-5 minutes. 3)Stain slides with May- Grunwald stain solution for 3 minutes. (Note :Staining time may vary depending on concentration of stain, determine optimal staining time with a trial slide before proceeding.) 4)Add equal amount of distilled water, tilt to mix and stain 1 minute Staining . A drop of stain can be added before the cover slip is placed or after having examined the unstained preparation. The stain will diffuse and then you can examine it. The . iodine . will stain the organisms a . dark orange brown . color. If you use new . methylene blue . instead, you will see organisms . contrasted against a blue.
Chapter 2-2a: Lab Techniques: Slide Preparation and Stains 2-2a-3 CHAPTER 2-2a LAB TECHNIQUES: SLIDE PREPARATION AND STAINS Figure 1. Polytrichum juniperinum leaf cross section using a cryostat and displaying natural colors. Photo courtesy of John Hribljan. Preparing the Specimen Fresh specimens are the most fun to work with. The The Ziehl-Neelsen stain, also known as the acid-fast stain, widely used differential staining procedure. The Ziehl - Neelsen stain was first described by two German doctors; Franz Ziehl (1859 to 1926), a bacteriologist and Friedrich Neelsen (1854 to 1894) a pathologist. In this type some bacteria resis Procedure. 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. Let stand for 2-3 minutes Clean grease-free glass slide, staining rack, blotting paper, immersion oil, and microscope. Procedure 1. 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.
Staining Procedure: Fill up two Coplin jars or wide-mouth bottles: Field Stain A (Blue stain). Field Stain B (Red stain). Make blood smear on a clean glass slide and it is dried in the air. Fix in methanol for one minute or get Spray 'Easyfix'. Dry in the air. Dip fixed smear to Field Stain B (Red Stain) for 5 to 6 seconds Experiment on Blood Film: Preparation: is based on their size and shape of nucleus and differential staining of nucleus and cytoplasm with Giemsa or Leishman stain. Technique: Prepare blood film following standard technique and stain with Giemsa or Leishman stain. Procedure: Put 2 ml blood in the oxalated vial and shake for 3 minutes.
information possible. If you have questions about patient preparation for any test, refer to our Test Directory or contact Client Services at 440-329-7863 for further assistance. Fasting requirements: o For the majority of test(s) performed on serum, plasma or whole blood, a fasting specimen is preferred. Fa Part 5: Gram Stain Procedure . Place slide with heat fixed smear on staining tray. Gently flood smear with crystal violet and let stand for 1 minute. Tilt the slide slightly and gently rinse with tap water or distilled water using a wash bottle. Gently flood the smear with Gram's iodine and let stand for 1 minute
10 Collection and Preparation of Material for Cytodiagnosis Accurate interpretation of cellular material is dependent on the following factors: Methods of specimen collection. Fixation and fixatives. Preservation of fluid specimens prior to processing. Preparation of material for microscopic examination. Staining and mounting of the cell sample How to Make a Blood Smear STEP ONE: Place clean glass slide on a flat surface. Add one small drop of blood to one end. STEP TWO: Take another clean slide, and holding at an angle of about 45 deg, touch the blood with one end of the slide so the blood runs along the edge of the slide by capillary action. Pus The DxH Slidemaker Stainer II (SMS II) prepares slides automatically based on orders received from your LIS. Select your own criteria for blood film preparation and define your own staining protocols. Receive high-quality slides on first pass, speeding the decision-making process and aiding cell classification and diagnosis Proceed to staining. 01 PROTOCOL: Method I - Frozen samples (whole animal or dissected tissue) 02 03 Fixation: Whole animal - Perfuse the animal with warm saline solution to flush the blood out of vasculature and immediately follow this by perfusion with freshly prepared 4% formaldehyde Papanicolaou stain (also Papanicolaou's stain or PAP stain) is the most important stain utilized in the practice of Cytopathology. It is a polychromatic stain containing multiple dyes to differentially stain various components of the cells. This technique was developed by George Papanicolaou, the father of Cytopathology
This procedure is known as a blood film. Initially, the blood smear test was done manually using a microscope. But, with the advancement of technology, automated digital systems have become available that help to analyze blood smears in a more efficient manner. Blood smear test is a low risk procedure. And no special preparation is needed for. Blood Cells has been written with both the practising haematologist and the trainee in mind. It aims to provide a guide for use in the diagnostic haematology laboratory, covering methods of collection of blood specimens, blood film preparation and staining, the principles of manual and automated blood counts and the assessment of the morphological features of blood cells 1. Microscopy. a) Thick and thin blood smear study. Thick and thin blood smear study is the gold standard method for malaria diagnosis. The procedure follows these steps: collection of peripheral blood, staining of smear with Giemsa stain and examination of red blood cells for malaria parasites under the microscope. Thick smear
T hen plac e the blood drop 1cm from the end of the slide. P roceeding with the 45 degree wedge or push slide technique used in manual and automated environments, creates a monolayer blood smear. This is done i n a smooth and quick motion. Fixation, staining, washing and air drying are quickly commenced 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-Giemsa are universally used in haematology. A Romanowsky stain is any stain combination consisting o Method. Cover blood film completely with the Leishman stain using a Pasteur pipette and wait for 2-3 minutes. Approximately 3 mL of stain is required to stain a single slide. Add an equal volume of phosphate buffer onto the slide (stain:buffer ratio = 1:1). Use a Pasteur pipette to blow gently over the slide to completely mix the stain and buffer Peripheral Blood Mononuclear Cell (PBMC) Isolation and Red Cell Lysis Procedures Introduction: Leukocytes are the most commonly analyzed cells in flow cytometry. Leukocytes can be obtained from whole blood and a variety of tissues, such as spleen, lymph node, bone marrow and thymus Submit all blood smears, perfect or otherwise, as areas of the film may be suitable for examination. Collect blood in an EDTA tube and make the smears when back at the clinic. Use clean, high-quality microscope slides. Aim for a blood droplet size of 4mm diameter. Optimise spreading speed for length and a good feathered edge
Enables both the haematologist and laboratory scientist to identify blood cell features, from the most common to the more obscure; Provides essential information on methods of collection, blood film preparation and staining, together with the principles of manual and automated blood count staining intensity is made by visualizing good color contrast between the cell nucleus and cytoplasm. The most important aspect of staining and slide preparation is to be able to visualize good nuclear and cytoplasmic detail. Without good slide preparation and staining, a reliable cytologic interpretation cannot be made. INTERPRETATIO Figure 1: Hematology staining of a blood smear. Figure 2: Hematology staining of blood smears at pH 6.4 (A), 6.8 (B), and 7.2 (C). Figure 3: Histology staining of lymph node (A), gastric mucosa infected with H. pylori (B), and Iliac crest biopsies (C & D). Figure 5: Bacteriology staining of infected blood. Malaria-infected blood-Gametocyte. A blood film allows the evaluation of white blood cells (WBCs, leucocytes), red blood cells (RBCs, erythrocytes), and platelets (thrombocytes). These cell are produced and mature in the bone marrow and are released into the bloodstream when needed. WBCs' main function is to fight infection, while RBCs carry oxygen to the whole of the body Perl stain; Myeloperoxidase (MPO) This is an enzyme that is located in the granules of both the myeloid and monocytic cells. As a stain, it is positive in cells of granulocyte series. Its activity increases with the maturation of myeloid cells while the immature myeloid cells tend to have very little to no staining
Routine H&E staining and special stains play a critical role in tissue-based diagnosis or research. By colouring otherwise transparent tissue sections, these stains allow highly trained pathologists and researchers to view, under a microscope, tissue morphology (structure) or to look for the presence or prevalence of particular cell types, structures or even microorganisms such as bacteria Processing cytology samples in the laboratory. Processing cytology samples involves reception of the specimen and request form, preparation of slides for microscopic examination, staining, screening and reporting the slides. All these processes are subject to quality control and quality assurance measures
The document covers the methods for the staining procedures commonly used in clinical microbiology laboratories for the identification of pathogens and dyes/ stains used for the differentiation of blood cells, for example, methylene blue stain used for white blood cell (WBC) differentiation. The dyes/ stains are covered in the appendix Malaria Test Procedures Blood Smears. As mentioned above, in order to conduct this test, two types of smears - Thick (greater volume of blood would mean higher chances of detecting malarial parasites) and Thin (fewer blood cells present in the sample allow the identification of the type of Plasmodium species causing the infection in the. 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.
Before staining and observing a microbe under a microscope, a smear must be prepared. 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. Figure 3-4 demonstrates smear preparation A Peripheral Blood Smear Examination is a procedure, which involves spreading a drop of blood thinly onto a glass slide. It is then treated with a special stain and studied under a microscope to examine the blood cells; The drop of blood used for a Blood Smear test contains numerous red blood cells (RBCs), white blood cells (WBCs), and platelets TECHNICAL DOCUMENT Handbook on TB laboratory diagnostic methods for the EU 1 Background and introduction Francis Drobniewski Tuberculosis (TB) is a major cause of morbidity and mortality in Europe. High-quality laboratory diagnosis of TB i Therefore, examination of a thick blood film is recommended. With a thick blood film, the red cells are approximately 6 - 20 layers thick which results in a larger volume of blood being examined. Field's stain method for THICK blood films The method recommended for staining thick blood films by the Hospital for Tropical Diseases is Field's. Microscopy of peripheral blood thin and thick films remains the reference for malaria diagnosis. Although Giemsa staining is most commonly used, the Leishman staining method provides better visualization of the nuclear chromatin pattern of cells. It is less well known whether accuracy of parasitaemia assessment is equally accurate with the latter method
Transfer the culture to a centrifuge tube and spin at 500xg for 5 minutes. Remove the supernatant and re-suspend the cells in 5-10ml of hypotonic 0.075M KCl (GIBCO Cat. No. 10575-090). Incubate at 37ºC for 10-12 minutes. Spin at 500xg for 5 minutes. Remove the supernatant, agitate the cellular sediment and add drop-by-drop 5-10ml of fresh, ice. The blood smear does not have to be positive for blood parasites. Good color differentiation of red and white cells is an indication of a good quality stain. Results of Q.C. smears for blood parasites are recorded on the QC Record Sheet for Giemsa Staining. This record is maintained in the QC Log Book. STAINING PROCEDURE A. Thin Blood Smears 1 A thick blood smear is a drop of blood on a glass slide. Thick blood smears are most useful for detecting the presence of parasites, because they examine a larger sample of blood. (Often there are few parasites in the blood at the time the test is done.) A thin blood smear is a drop of blood that is spread across a large area of the slide 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