This document provides information on preparing and staining peripheral blood smears (PBS). It discusses how to make a wedge blood smear using the correct technique and equipment. It also describes how to evaluate a quality smear and identify common causes of poor smears. The document outlines the staining process for PBS, including the history and components of Romanowsky staining methods like Wright-Giemsa and May-Grunwald Giemsa. Factors that can influence staining and cause faulty results are discussed. Finally, it provides guidance on examining a PBS under the microscope, including evaluating red blood cells, white blood cells, platelets, and identifying any parasites.
This document provides information on staining blood films and smears. It discusses the different types of stains used including Romanowsky stains like Leishman stain, Giemsa stain, Wright stain, and Field stain. Specimens should be collected in EDTA and smears prepared within an hour then fixed in methanol or ethanol to preserve cell morphology before staining. Romanowsky stains use methylene blue and eosin dyes to reveal subtle differences in cell structures and components.
Romanowsky stains are commonly used to stain blood films and identify blood components. Some key Romanowsky stains discussed in the document include Leishman, Giemsa, Wright's, Field, Jenner, and JSB stains. These stains involve using dyes like methylene blue and eosin in specific combinations and concentrations to differentially stain structures in blood films based on their chemical properties. Proper staining technique and protocols are outlined to clearly identify red blood cells, white blood cells, parasites, and other components when examining stained blood films under a microscope.
This document discusses the peripheral blood smear examination. It begins by outlining the role of peripheral blood examination, which includes evaluating anemia, thrombocytopenia/thrombocytosis, identifying abnormal cells, and detecting infections. It then describes the proper collection of blood in EDTA tubes and various color-coded tubes. The document proceeds to explain the different techniques for preparing blood smears, including the wedge, cover slip, and spun smear methods. Finally, it outlines the staining, microscopic examination, identification of white blood cells, red blood cell abnormalities, and other findings commonly seen on peripheral blood smears.
This document discusses the determination of red blood cell indices including packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC). It describes the macrohematocrit or Wintrobe tube method for determining PCV which involves centrifuging a blood sample and measuring the ratio of packed red cells to total blood volume. Formulas are provided for calculating MCV from PCV and RBC count, MCH from hemoglobin and RBC count, and MCHC from hemoglobin and PCV. Normal ranges are listed for each index.
This document discusses the preparation and staining of blood films, which is an important investigation in hematology. It describes how Romanowsky stains like Wright's, Leishman, Giemsa, and Jenner's stains are commonly used to differentially stain components of blood cells. Specifically, it provides details on the preparation and staining procedures for Leishman's stain and Giemsa's stain, the most frequently used stains. An optimal pH of 6.8 is recommended for staining all components when using Romanowsky stains.
I have listed out the LE cells structure and Microscopical examinaton of LE CELLS, Difference between tart cells and le cells, clinical symptoms and diagnostic procedure.
This document discusses different types of anticoagulants used in blood collection and their properties. It describes how anticoagulants prevent blood from clotting by binding with calcium ions which are essential for the coagulation process. The most commonly used anticoagulant is EDTA as it best preserves cell morphology. Other anticoagulants mentioned include citrate, heparin, and oxalates. The document provides details on the preparation and use of different anticoagulants to optimize their anticoagulant effects without damaging blood cells.
This document discusses various anticoagulants used in hematology. It describes the characteristics anticoagulants should have and provides details on commonly used anticoagulants including EDTA, oxalates, heparin, sodium citrate, and sodium fluoride/potassium oxalate mixtures. The anticoagulants are classified as calcium chelators or non-calcium chelators and the mechanisms of action, concentrations, advantages, and disadvantages of each type are outlined.
This document provides information on staining blood films and smears. It discusses the different types of stains used including Romanowsky stains like Leishman stain, Giemsa stain, Wright stain, and Field stain. Specimens should be collected in EDTA and smears prepared within an hour then fixed in methanol or ethanol to preserve cell morphology before staining. Romanowsky stains use methylene blue and eosin dyes to reveal subtle differences in cell structures and components.
Romanowsky stains are commonly used to stain blood films and identify blood components. Some key Romanowsky stains discussed in the document include Leishman, Giemsa, Wright's, Field, Jenner, and JSB stains. These stains involve using dyes like methylene blue and eosin in specific combinations and concentrations to differentially stain structures in blood films based on their chemical properties. Proper staining technique and protocols are outlined to clearly identify red blood cells, white blood cells, parasites, and other components when examining stained blood films under a microscope.
This document discusses the peripheral blood smear examination. It begins by outlining the role of peripheral blood examination, which includes evaluating anemia, thrombocytopenia/thrombocytosis, identifying abnormal cells, and detecting infections. It then describes the proper collection of blood in EDTA tubes and various color-coded tubes. The document proceeds to explain the different techniques for preparing blood smears, including the wedge, cover slip, and spun smear methods. Finally, it outlines the staining, microscopic examination, identification of white blood cells, red blood cell abnormalities, and other findings commonly seen on peripheral blood smears.
This document discusses the determination of red blood cell indices including packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC). It describes the macrohematocrit or Wintrobe tube method for determining PCV which involves centrifuging a blood sample and measuring the ratio of packed red cells to total blood volume. Formulas are provided for calculating MCV from PCV and RBC count, MCH from hemoglobin and RBC count, and MCHC from hemoglobin and PCV. Normal ranges are listed for each index.
This document discusses the preparation and staining of blood films, which is an important investigation in hematology. It describes how Romanowsky stains like Wright's, Leishman, Giemsa, and Jenner's stains are commonly used to differentially stain components of blood cells. Specifically, it provides details on the preparation and staining procedures for Leishman's stain and Giemsa's stain, the most frequently used stains. An optimal pH of 6.8 is recommended for staining all components when using Romanowsky stains.
I have listed out the LE cells structure and Microscopical examinaton of LE CELLS, Difference between tart cells and le cells, clinical symptoms and diagnostic procedure.
This document discusses different types of anticoagulants used in blood collection and their properties. It describes how anticoagulants prevent blood from clotting by binding with calcium ions which are essential for the coagulation process. The most commonly used anticoagulant is EDTA as it best preserves cell morphology. Other anticoagulants mentioned include citrate, heparin, and oxalates. The document provides details on the preparation and use of different anticoagulants to optimize their anticoagulant effects without damaging blood cells.
This document discusses various anticoagulants used in hematology. It describes the characteristics anticoagulants should have and provides details on commonly used anticoagulants including EDTA, oxalates, heparin, sodium citrate, and sodium fluoride/potassium oxalate mixtures. The anticoagulants are classified as calcium chelators or non-calcium chelators and the mechanisms of action, concentrations, advantages, and disadvantages of each type are outlined.
special and routine stains in haematology 1Dr.SHAHID Raza
The document discusses various routine and special stains used in hematology. Routine stains like Leishman, Giemsa, and Wright stains are used to stain peripheral blood films and differentiate blood cells. Special stains require additional processing but can identify characteristics not seen with routine stains, such as periodic acid Schiff stain which detects carbohydrates like glycogen by oxidizing glycol groups and producing a red reaction. Proper staining techniques such as fixation, washing, and timing are important for preparing clear blood smears and accurately identifying blood components.
Dr. Pandian M describes the procedure for performing a platelet count. Platelets serve important hemostatic functions and their normal range is 1.5-4 lakhs/cumm. The procedure involves mixing blood with a diluting fluid in a Neubauer chamber, then counting platelets in grid squares under a microscope. For the sample, 40 platelets were counted in 1/50 mm3, indicating a platelet count of 2 lakhs/mm3 of blood, within the normal range. Abnormally high or low platelet counts can occur due to various bone marrow and other disorders.
An immature red blood cell without a nucleus, having a granular or reticulated appearance when suitably stained.
Reticulocytes are the immature RBC that contain nucleus.
They are originally seen at the site of their formation i.e. bone marrow. They take 2-3 (lays for maturation only about 1-2% of circulating RBCs are Reticulocytes.
This document provides instructions for preparing and staining a peripheral blood smear. It describes how to make a thin blood film using the wedge technique and a thick blood film for diagnosing parasites. Potential sources of error in film preparation are outlined. The document then explains the staining process for Leishman's stain and Giemsa stain, including stain preparation, application to blood films, and differentiation to distinguish cells under the microscope. The stains are used to identify cells after a blood film has been prepared.
This document provides information on three hematology stains:
Leishman's stain, reticulocyte stain, and iron stain. It describes the composition, procedure, and results for Leishman's stain. For reticulocyte stain, it discusses how reticulocytes contain RNA that stains gray-blue and the procedure uses supravital staining. Iron stain demonstrates ferric iron and ferritin using Prussian blue reaction and involves fixation, staining in potassium ferrocyanide and hydrochloric acid, and counterstaining with nuclear fast red.
The osmotic fragility test measures the resistance of red blood cells to lysis when exposed to saline solutions of varying concentrations. Small amounts of blood are mixed with buffered saline solutions and the percentage of lysed red blood cells is determined. Increased osmotic fragility indicates conditions that cause spherical red blood cells like hereditary spherocytosis, while decreased fragility occurs in iron deficiency anemia and thalassemia where cells are flattened. The test is a simple way to evaluate red blood cell shape and membrane stability.
This document provides guidelines for peripheral blood smear examination, including slide preparation, staining, and microscopic evaluation. Key points include:
- Blood smears are used to evaluate anemia, infections, and abnormal cells. Three steps are involved in making blood films: preparation, fixation, and staining.
- Under the microscope, a preliminary low and high power scan examines staining quality and cell distribution before a 100-cell differential count at oil immersion.
- RBCs, WBCs, platelets, and parasites are examined for morphologies, counts, and any abnormalities. Terms like pancytopenia and bicytopenia are defined.
- Detailed guidelines are given for examining and reporting R
The LE cell demonstration document describes the LE cell, which is a neutrophil that has phagocytosed nuclear material coated with antinuclear antibodies, a characteristic of lupus erythematosus. It discusses several methods for demonstrating LE cells in blood samples, including using clotted blood, defibrinated blood, or the rotary method. The rotary method involves adding glass beads to heparinized blood and rotating at 50rpm for 30 minutes at 37 degrees Celsius before preparing buffy coat smears to identify LE cells.
This document discusses reticulocytes, which are immature red blood cells that spend 1-3 days maturing in the bone marrow before circulating in the bloodstream. Key points include:
- Reticulocytes have residual RNA that allows them to be stained and counted to assess bone marrow activity.
- An elevated reticulocyte count indicates the bone marrow is responding to anemic stress by increasing red blood cell production.
- The reticulocyte count is used to evaluate bone marrow function and health in conditions like anemia, blood loss, or following chemotherapy/radiation.
Morphological abnormality of white blood cellNAZAR ABU-DULLA
This presentation describe the normal WBC normal and abnormal shape.
it can also describe the maturation of different WBC and reactivity of the WBC different infection
This document discusses automation in hematology. It begins by outlining the necessity for automation in cell counting, diagnosing various blood conditions, and performing multiple tests on a single platform. The document then covers the advantages and disadvantages of automation, including increased speed and accuracy versus high costs. It describes the various principles used in automated hematology analyzers, such as electrical impedance, optical light scattering, and flow cytometry. Finally, it provides details on specific analyzers like the Pentra ES 60 and Pentra DF Nexus.
Bone marrow aspirate&biopsy preparationMalak Salam
This document discusses bone marrow aspiration and biopsy procedures. Key points:
- Bone marrow biopsy is important for diagnosing blood diseases and may be the only way to make a correct diagnosis. Marrow can be obtained repeatedly by needle aspiration.
- The iliac crests are the preferred sites for aspiration in adults and children. The sternum should not be used in children due to risk of injury.
- Proper needles, anticoagulants, and techniques are required to safely aspirate marrow and prepare diagnostic films and samples. Cell counts and differentials on aspirated marrow provide important diagnostic information.
The document discusses different methods of blood collection including capillary, venous, and arterial blood. It describes the equipment and procedures used to collect blood via these methods and the tests that each type is suited for. The document also outlines the various anticoagulants used in blood collection tubes, including EDTA, citrate, heparin, oxalate, and their mechanisms and uses. Finally, it addresses the order that tubes should be drawn in and storage of blood in blood banks.
This document discusses quality assurance in haematology. It defines quality and introduces the concepts of quality control and quality assurance. Quality control aims to minimize errors through statistical sampling and verification of consistent performance. Quality assurance ensures reliable test results through adherence to standards within and outside the laboratory. This includes internal quality control, external quality assessment, and standardization using reference materials and methods. Several examples are provided of potential pre-analytical errors in sample collection, transport, and handling that can affect test results. Adherence to proper procedures is emphasized to avoid issues like hemolysis, clotting, and dilution.
Platelet function tests.pptx 2.pptx finalAnupam Singh
This document summarizes platelet function testing. It discusses how platelets are formed from megakaryocytes in the bone marrow and circulate in the bloodstream. The major platelet function tests are platelet aggregometry, flow cytometry, and point-of-care tests like the impact cone and plate analyzer and thromboelastography. These tests are used to diagnose platelet disorders and monitor antiplatelet therapy. The document also briefly discusses platelet-derived microparticles and microRNAs, which can provide information about platelet activation and signaling.
The document discusses different methods of blood collection including capillary, venous, and arterial blood. It describes various anticoagulants used in blood collection tubes and their purposes, such as EDTA for cell counts and citrate for coagulation studies. The document also outlines the proper order for drawing blood into collection tubes and the use of blood banks for storing blood components.
This document describes the osmotic fragility test procedure and its use in evaluating red blood cell disorders. It involves incubating blood samples in serially diluted saline solutions and analyzing hemolysis. Abnormally increased or decreased fragility can indicate conditions like hereditary spherocytosis or iron deficiency anemia respectively. A modified test called NESTROFT is also described, which is useful for screening for beta thalassemia trait in areas without automated analyzers.
Peripheral blood smear examination plays an important role in the evaluation of various blood disorders. A good peripheral smear should be prepared using the wedge or coverslip technique to obtain an even distribution of red blood cells. The smear is then stained using the Romanowsky method which involves fixing the cells using methanol followed by staining with Giemsa stain. During examination, red blood cells, white blood cells, platelets and any abnormal cells or inclusions are evaluated under the microscope. Changes in the size, shape, color and structural features of red blood cells can provide clues to underlying hematological conditions.
This document discusses the peripheral blood smear examination process. It begins by outlining the role of peripheral blood examination, including evaluation of anemia, thrombocytopenia/thrombocytosis, and identification of abnormal cells and infections. It then describes the collection of blood in EDTA tubes, and preparation of blood smears using the wedge, cover slip, and spun techniques. Finally, it details the staining process using Leishman's stain, Field's stain, and automated slide stainers, and provides an overview of examining blood smears under the microscope.
special and routine stains in haematology 1Dr.SHAHID Raza
The document discusses various routine and special stains used in hematology. Routine stains like Leishman, Giemsa, and Wright stains are used to stain peripheral blood films and differentiate blood cells. Special stains require additional processing but can identify characteristics not seen with routine stains, such as periodic acid Schiff stain which detects carbohydrates like glycogen by oxidizing glycol groups and producing a red reaction. Proper staining techniques such as fixation, washing, and timing are important for preparing clear blood smears and accurately identifying blood components.
Dr. Pandian M describes the procedure for performing a platelet count. Platelets serve important hemostatic functions and their normal range is 1.5-4 lakhs/cumm. The procedure involves mixing blood with a diluting fluid in a Neubauer chamber, then counting platelets in grid squares under a microscope. For the sample, 40 platelets were counted in 1/50 mm3, indicating a platelet count of 2 lakhs/mm3 of blood, within the normal range. Abnormally high or low platelet counts can occur due to various bone marrow and other disorders.
An immature red blood cell without a nucleus, having a granular or reticulated appearance when suitably stained.
Reticulocytes are the immature RBC that contain nucleus.
They are originally seen at the site of their formation i.e. bone marrow. They take 2-3 (lays for maturation only about 1-2% of circulating RBCs are Reticulocytes.
This document provides instructions for preparing and staining a peripheral blood smear. It describes how to make a thin blood film using the wedge technique and a thick blood film for diagnosing parasites. Potential sources of error in film preparation are outlined. The document then explains the staining process for Leishman's stain and Giemsa stain, including stain preparation, application to blood films, and differentiation to distinguish cells under the microscope. The stains are used to identify cells after a blood film has been prepared.
This document provides information on three hematology stains:
Leishman's stain, reticulocyte stain, and iron stain. It describes the composition, procedure, and results for Leishman's stain. For reticulocyte stain, it discusses how reticulocytes contain RNA that stains gray-blue and the procedure uses supravital staining. Iron stain demonstrates ferric iron and ferritin using Prussian blue reaction and involves fixation, staining in potassium ferrocyanide and hydrochloric acid, and counterstaining with nuclear fast red.
The osmotic fragility test measures the resistance of red blood cells to lysis when exposed to saline solutions of varying concentrations. Small amounts of blood are mixed with buffered saline solutions and the percentage of lysed red blood cells is determined. Increased osmotic fragility indicates conditions that cause spherical red blood cells like hereditary spherocytosis, while decreased fragility occurs in iron deficiency anemia and thalassemia where cells are flattened. The test is a simple way to evaluate red blood cell shape and membrane stability.
This document provides guidelines for peripheral blood smear examination, including slide preparation, staining, and microscopic evaluation. Key points include:
- Blood smears are used to evaluate anemia, infections, and abnormal cells. Three steps are involved in making blood films: preparation, fixation, and staining.
- Under the microscope, a preliminary low and high power scan examines staining quality and cell distribution before a 100-cell differential count at oil immersion.
- RBCs, WBCs, platelets, and parasites are examined for morphologies, counts, and any abnormalities. Terms like pancytopenia and bicytopenia are defined.
- Detailed guidelines are given for examining and reporting R
The LE cell demonstration document describes the LE cell, which is a neutrophil that has phagocytosed nuclear material coated with antinuclear antibodies, a characteristic of lupus erythematosus. It discusses several methods for demonstrating LE cells in blood samples, including using clotted blood, defibrinated blood, or the rotary method. The rotary method involves adding glass beads to heparinized blood and rotating at 50rpm for 30 minutes at 37 degrees Celsius before preparing buffy coat smears to identify LE cells.
This document discusses reticulocytes, which are immature red blood cells that spend 1-3 days maturing in the bone marrow before circulating in the bloodstream. Key points include:
- Reticulocytes have residual RNA that allows them to be stained and counted to assess bone marrow activity.
- An elevated reticulocyte count indicates the bone marrow is responding to anemic stress by increasing red blood cell production.
- The reticulocyte count is used to evaluate bone marrow function and health in conditions like anemia, blood loss, or following chemotherapy/radiation.
Morphological abnormality of white blood cellNAZAR ABU-DULLA
This presentation describe the normal WBC normal and abnormal shape.
it can also describe the maturation of different WBC and reactivity of the WBC different infection
This document discusses automation in hematology. It begins by outlining the necessity for automation in cell counting, diagnosing various blood conditions, and performing multiple tests on a single platform. The document then covers the advantages and disadvantages of automation, including increased speed and accuracy versus high costs. It describes the various principles used in automated hematology analyzers, such as electrical impedance, optical light scattering, and flow cytometry. Finally, it provides details on specific analyzers like the Pentra ES 60 and Pentra DF Nexus.
Bone marrow aspirate&biopsy preparationMalak Salam
This document discusses bone marrow aspiration and biopsy procedures. Key points:
- Bone marrow biopsy is important for diagnosing blood diseases and may be the only way to make a correct diagnosis. Marrow can be obtained repeatedly by needle aspiration.
- The iliac crests are the preferred sites for aspiration in adults and children. The sternum should not be used in children due to risk of injury.
- Proper needles, anticoagulants, and techniques are required to safely aspirate marrow and prepare diagnostic films and samples. Cell counts and differentials on aspirated marrow provide important diagnostic information.
The document discusses different methods of blood collection including capillary, venous, and arterial blood. It describes the equipment and procedures used to collect blood via these methods and the tests that each type is suited for. The document also outlines the various anticoagulants used in blood collection tubes, including EDTA, citrate, heparin, oxalate, and their mechanisms and uses. Finally, it addresses the order that tubes should be drawn in and storage of blood in blood banks.
This document discusses quality assurance in haematology. It defines quality and introduces the concepts of quality control and quality assurance. Quality control aims to minimize errors through statistical sampling and verification of consistent performance. Quality assurance ensures reliable test results through adherence to standards within and outside the laboratory. This includes internal quality control, external quality assessment, and standardization using reference materials and methods. Several examples are provided of potential pre-analytical errors in sample collection, transport, and handling that can affect test results. Adherence to proper procedures is emphasized to avoid issues like hemolysis, clotting, and dilution.
Platelet function tests.pptx 2.pptx finalAnupam Singh
This document summarizes platelet function testing. It discusses how platelets are formed from megakaryocytes in the bone marrow and circulate in the bloodstream. The major platelet function tests are platelet aggregometry, flow cytometry, and point-of-care tests like the impact cone and plate analyzer and thromboelastography. These tests are used to diagnose platelet disorders and monitor antiplatelet therapy. The document also briefly discusses platelet-derived microparticles and microRNAs, which can provide information about platelet activation and signaling.
The document discusses different methods of blood collection including capillary, venous, and arterial blood. It describes various anticoagulants used in blood collection tubes and their purposes, such as EDTA for cell counts and citrate for coagulation studies. The document also outlines the proper order for drawing blood into collection tubes and the use of blood banks for storing blood components.
This document describes the osmotic fragility test procedure and its use in evaluating red blood cell disorders. It involves incubating blood samples in serially diluted saline solutions and analyzing hemolysis. Abnormally increased or decreased fragility can indicate conditions like hereditary spherocytosis or iron deficiency anemia respectively. A modified test called NESTROFT is also described, which is useful for screening for beta thalassemia trait in areas without automated analyzers.
Peripheral blood smear examination plays an important role in the evaluation of various blood disorders. A good peripheral smear should be prepared using the wedge or coverslip technique to obtain an even distribution of red blood cells. The smear is then stained using the Romanowsky method which involves fixing the cells using methanol followed by staining with Giemsa stain. During examination, red blood cells, white blood cells, platelets and any abnormal cells or inclusions are evaluated under the microscope. Changes in the size, shape, color and structural features of red blood cells can provide clues to underlying hematological conditions.
This document discusses the peripheral blood smear examination process. It begins by outlining the role of peripheral blood examination, including evaluation of anemia, thrombocytopenia/thrombocytosis, and identification of abnormal cells and infections. It then describes the collection of blood in EDTA tubes, and preparation of blood smears using the wedge, cover slip, and spun techniques. Finally, it details the staining process using Leishman's stain, Field's stain, and automated slide stainers, and provides an overview of examining blood smears under the microscope.
The document provides information on blood films (blood smears) including:
- Blood films allow examination of blood cells under a microscope and are an important diagnostic tool.
- They are prepared by smearing a blood sample on a slide and staining it.
- Examination of thin and thick blood films can detect hematological disorders and blood parasites like malaria.
- Proper preparation, fixation, and staining are required to preserve cell morphology and obtain diagnostic information from blood films.
The document discusses various hematological investigations and artifacts. It describes the process of a differential count, which involves examining a peripheral blood smear under a microscope to determine percentages of different white blood cells. The blood smear can also reveal abnormal red blood cell and white blood cell morphologies. The document then discusses the steps to make a good blood smear, including using a clean slide and proper angulation and force when spreading the blood. Potential artifacts from improper smear preparation or staining are also described. The document concludes with discussing various hematological tests including complete blood count, erythrocyte sedimentation rate, coagulation tests, and factors that could affect the results.
This document provides an overview of peripheral blood smear examination. Key points include:
- A peripheral smear can provide information about red blood cells, white blood cells, and platelets to help diagnose anemias and other hematological disorders.
- Proper collection, staining, and microscopic evaluation are important for accurate results. Common stains include Leishman's, Giemsa, and Wright stains.
- Features of red blood cells like size, shape, color, and inclusions provide clues to different anemias. White blood cell differentials help identify infections and disorders.
- Artifacts can occur but proper technique and examination of multiple fields helps distinguish real from artifactual findings.
This document provides instructions for examining blood samples. It describes the components of blood and how to collect blood through either capillary or venous methods. Detailed procedures are provided for making wet blood preparations, thick blood films, and thin blood films in order to examine blood cells and detect parasites. Quality controls and common problems in blood film preparation are also discussed.
This document provides information on how to perform an erythrocyte sedimentation rate (ESR) test. It begins by explaining that ESR measures the rate at which red blood cells sediment in one hour, and though nonspecific, an increased ESR can indicate infection, inflammation or malignancy. It then notes some limitations before describing the basic principles behind ESR, including how plasma proteins promote rouleaux formation and faster sedimentation. Finally, it states that the Westergren method is the preferred technique for determining ESR over the Wintrobe method.
This document provides information on preparing blood smears with different staining methods. It discusses preparing thin and thick blood smears using various techniques like the slide method, cover glass method, and spin method for thin smears. For thick smears, a large drop of blood is spread on the slide. Various staining methods are described like Leishman stain, Giemsa stain, Wright stain, and Field stain. Proper preparation, fixation in methanol or ethanol, and staining are required to visualize blood cells and parasites under the microscope. Reticulocyte staining is also briefly mentioned.
This document provides information on preparing, fixing, and staining blood smears or peripheral blood films. It describes how to make thin and thick blood films using different methods like the slide, cover glass, and spin methods. Films need to be fixed using methanol to preserve cell morphology before staining. Various Romanowsky stains are commonly used like Leishman, Giemsa, Wright, Field, and Jenner stains. Proper staining techniques and precautions are outlined to produce high quality blood films for microscopic examination and identification of blood cells and parasites.
1. A peripheral blood film (PBF) is a thin layer of blood smeared on a microscope slide that is stained to allow examination of blood cells.
2. An ideal blood smear is uniformly thick, occupies the central portion of the slide, and tapers from thick at the head to thin at the tail.
3. Preparation involves using a spreader slide at a 40-45 degree angle to pull blood into a feathered tongue shape from a drop of blood on the slide.
Blood smear preparation, fixation, staining. Veterinary Clinical Examination ...Gansbaai SA
How to make a blood smear, including all steps: Preparation, fixation and staining. Veterinary clinical Examination for small and large animals. Methanol - Fixation. Additionaly basic information about blood.
The erythrocyte sedimentation rate (ESR) is a nonspecific screening test used to detect inflammation. It measures how fast red blood cells sediment in a tube in one hour. Faster sedimentation indicates more inflammation in the body from conditions like infections, cancers, and autoimmune diseases. The ESR is directly related to red cell mass and fibrinogen levels, and inversely related to plasma viscosity. It is an inexpensive and widely available initial screening test to monitor disease progression or remission.
The blood smear remains an important diagnostic tool for hematological disorders. It provides rapid information about disorders like anemia and thrombocytopenia. While automated analyzers are used, morphological examination of smears by trained technicians is needed for accurate diagnosis. Proper preparation of smears is important, involving placing a small blood drop on a slide and spreading it into a feathered wedge using another slide. Smears allow examination of red blood cell, white blood cell, and platelet morphology and differentials.
The blood smear document defines a blood smear, outlines its uses in diagnosing conditions like anemia and infections, and provides detailed instructions on preparing, staining, examining, and interpreting a blood smear slide. A well-prepared blood smear shows red and white blood cells with normal morphology, and is examined under microscopy to check for abnormalities and identify blood parasites or immature cells that can indicate conditions like infection or leukemia.
1. Wright's stain is a Romanowsky-type stain used to stain peripheral blood smears. It works by staining basic cell components pink/red and acidic components blue/purple.
2. The staining process involves overlaying a blood smear slide with Wright's stain for 2 minutes, then Giemsa solution for 4 minutes. The slide is then washed and allowed to dry.
3. Proper pH control is important for correct staining - a pH that is too acidic or alkaline can lead to poorly differentiated staining of cell structures.
The document discusses the importance of blood smears in diagnosing various hematological disorders. It states that blood smears provide a rapid and inexpensive way to examine blood cell morphology and detect abnormalities. While automated analyzers are used more often, blood smears remain an important diagnostic tool, especially when clinical findings require further investigation or there are discrepancies with previous test results. The document also provides guidance on properly preparing blood smears to obtain diagnostic quality samples.
This document describes several instruments used in pathology laboratories. It discusses urinometers for measuring urine specific gravity, Sahli's haemoglobinometer for estimating haemoglobin levels, and Esbach's albuminometer for testing albumin in urine. It also covers Neubauer's chamber and haemocytometers for cell counting, pipettes for blood cell counts, Westergren's tube for erythrocyte sedimentation rate testing, and vials for blood sample collection. Other instruments mentioned include thermometers, cassettes, microtome knives, pipettes, and syringes.
The document discusses different staining techniques used in hematology to differentiate blood cells. It describes routine stains like Leishman, Giemsa, Wright, and Jenner stains which use combinations of basic and acidic dyes to stain cell nuclei and cytoplasm. Proper preparation of thin and thick blood films, including fixation in methanol, is outlined. The principles, procedures, and interpretations of common Romanowsky stains are explained in detail.
Similar to Peripheral smear staining and morphology (20)
Four FDA-approved PD-L1 immunohistochemical assays use different antibodies to identify expression of PD-L1 protein in tumor and immune cells. Tumor proportion score (TPS) and combined positive score (CPS) are used to quantify PD-L1 staining and determine patient eligibility for various FDA-approved immune checkpoint inhibitor therapies targeting PD-1 and PD-L1 in cancers like NSCLC, bladder cancer, and gastric cancer. While TPS only considers tumor cell staining, CPS provides a more comprehensive score by including positive tumor and immune cells. Correct interpretation of PD-L1 immunohistochemistry requires attention to staining patterns, cell types, and cut-off thresholds specified for each companion diagnostic assay.
Genomics of bladder cancer and novel molecular taxonomy.pptxSudipta Naskar
Bladder cancers exhibit complex genotype with significant intratumoral heterogeneity. Molecular classification can provide insight to the more precise and individualized therapeutic approach
Two cases of Waldenstrӧm macroglobulinemia along with brief disease pathology. Waldenstrӧm macroglobulinemia is a rare B cell lymphoma involving bone marrow with IgM monoclonal gammopathy of any concentration
Molecular testing techniques in cytology specimensSudipta Naskar
Molecular testing techniques can be used on cytology specimens to facilitate cancer patient management. Fluorescence in situ hybridization (FISH) is well-suited for detecting genomic abnormalities in cytology specimens. FISH involves hybridizing fluorescent probes to target sequences to visualize locations. It can detect gains, losses, amplifications, and rearrangements. A variety of cytology specimens can be used for FISH, including smears, cell blocks, and liquid-based preparations. FISH has applications in detecting abnormalities in cancers like urothelial carcinoma, breast cancer, and lymphoma.
Study on utilization of antenatal care and outcome of pregnancy in a medical ...Sudipta Naskar
A study on Ante-natal care utilization by the mothers in Calcutta National Medical College & Hospital done by the 3rd Professional (Part I) M.B.B.S. students of Calcutta National Medical College under the guidance of the teachers of Community Medicine.
These lecture slides, by Dr Sidra Arshad, offer a quick overview of the physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar lead (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
6. Describe the flow of current around the heart during the cardiac cycle
7. Discuss the placement and polarity of the leads of electrocardiograph
8. Describe the normal electrocardiograms recorded from the limb leads and explain the physiological basis of the different records that are obtained
9. Define mean electrical vector (axis) of the heart and give the normal range
10. Define the mean QRS vector
11. Describe the axes of leads (hexagonal reference system)
12. Comprehend the vectorial analysis of the normal ECG
13. Determine the mean electrical axis of the ventricular QRS and appreciate the mean axis deviation
14. Explain the concepts of current of injury, J point, and their significance
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. Chapter 3, Cardiology Explained, https://www.ncbi.nlm.nih.gov/books/NBK2214/
7. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
TEST BANK For Community Health Nursing A Canadian Perspective, 5th Edition by...Donc Test
TEST BANK For Community Health Nursing A Canadian Perspective, 5th Edition by Stamler, Verified Chapters 1 - 33, Complete Newest Version Community Health Nursing A Canadian Perspective, 5th Edition by Stamler, Verified Chapters 1 - 33, Complete Newest Version Community Health Nursing A Canadian Perspective, 5th Edition by Stamler Community Health Nursing A Canadian Perspective, 5th Edition TEST BANK by Stamler Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Pdf Chapters Download Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Pdf Download Stuvia Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Study Guide Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Ebook Download Stuvia Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Questions and Answers Quizlet Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Studocu Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Quizlet Test Bank For Community Health Nursing A Canadian Perspective, 5th Edition Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Pdf Chapters Download Community Health Nursing A Canadian Perspective, 5th Edition Pdf Download Course Hero Community Health Nursing A Canadian Perspective, 5th Edition Answers Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Ebook Download Course hero Community Health Nursing A Canadian Perspective, 5th Edition Questions and Answers Community Health Nursing A Canadian Perspective, 5th Edition Studocu Community Health Nursing A Canadian Perspective, 5th Edition Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Pdf Chapters Download Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Pdf Download Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Study Guide Questions and Answers Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Ebook Download Stuvia Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Questions Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Studocu Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Quizlet Community Health Nursing A Canadian Perspective, 5th Edition Test Bank Stuvia
NVBDCP.pptx Nation vector borne disease control programSapna Thakur
NVBDCP was launched in 2003-2004 . Vector-Borne Disease: Disease that results from an infection transmitted to humans and other animals by blood-feeding arthropods, such as mosquitoes, ticks, and fleas. Examples of vector-borne diseases include Dengue fever, West Nile Virus, Lyme disease, and malaria.
Adhd Medication Shortage Uk - trinexpharmacy.comreignlana06
The UK is currently facing a Adhd Medication Shortage Uk, which has left many patients and their families grappling with uncertainty and frustration. ADHD, or Attention Deficit Hyperactivity Disorder, is a chronic condition that requires consistent medication to manage effectively. This shortage has highlighted the critical role these medications play in the daily lives of those affected by ADHD. Contact : +1 (747) 209 – 3649 E-mail : sales@trinexpharmacy.com
share - Lions, tigers, AI and health misinformation, oh my!.pptxTina Purnat
• Pitfalls and pivots needed to use AI effectively in public health
• Evidence-based strategies to address health misinformation effectively
• Building trust with communities online and offline
• Equipping health professionals to address questions, concerns and health misinformation
• Assessing risk and mitigating harm from adverse health narratives in communities, health workforce and health system
Cell Therapy Expansion and Challenges in Autoimmune DiseaseHealth Advances
There is increasing confidence that cell therapies will soon play a role in the treatment of autoimmune disorders, but the extent of this impact remains to be seen. Early readouts on autologous CAR-Ts in lupus are encouraging, but manufacturing and cost limitations are likely to restrict access to highly refractory patients. Allogeneic CAR-Ts have the potential to broaden access to earlier lines of treatment due to their inherent cost benefits, however they will need to demonstrate comparable or improved efficacy to established modalities.
In addition to infrastructure and capacity constraints, CAR-Ts face a very different risk-benefit dynamic in autoimmune compared to oncology, highlighting the need for tolerable therapies with low adverse event risk. CAR-NK and Treg-based therapies are also being developed in certain autoimmune disorders and may demonstrate favorable safety profiles. Several novel non-cell therapies such as bispecific antibodies, nanobodies, and RNAi drugs, may also offer future alternative competitive solutions with variable value propositions.
Widespread adoption of cell therapies will not only require strong efficacy and safety data, but also adapted pricing and access strategies. At oncology-based price points, CAR-Ts are unlikely to achieve broad market access in autoimmune disorders, with eligible patient populations that are potentially orders of magnitude greater than the number of currently addressable cancer patients. Developers have made strides towards reducing cell therapy COGS while improving manufacturing efficiency, but payors will inevitably restrict access until more sustainable pricing is achieved.
Despite these headwinds, industry leaders and investors remain confident that cell therapies are poised to address significant unmet need in patients suffering from autoimmune disorders. However, the extent of this impact on the treatment landscape remains to be seen, as the industry rapidly approaches an inflection point.
Osteoporosis - Definition , Evaluation and Management .pdfJim Jacob Roy
Osteoporosis is an increasing cause of morbidity among the elderly.
In this document , a brief outline of osteoporosis is given , including the risk factors of osteoporosis fractures , the indications for testing bone mineral density and the management of osteoporosis
Promoting Wellbeing - Applied Social Psychology - Psychology SuperNotesPsychoTech Services
A proprietary approach developed by bringing together the best of learning theories from Psychology, design principles from the world of visualization, and pedagogical methods from over a decade of training experience, that enables you to: Learn better, faster!
Does Over-Masturbation Contribute to Chronic Prostatitis.pptxwalterHu5
In some case, your chronic prostatitis may be related to over-masturbation. Generally, natural medicine Diuretic and Anti-inflammatory Pill can help mee get a cure.
#cALL# #gIRLS# In Dehradun ꧁❤8107221448❤꧂#cALL# #gIRLS# Service In Dehradun W...
Peripheral smear staining and morphology
1. P S -
S T A I N I N G
A N D
M O R P H O L O
G Y
S U D I PTA
2. What is a peripheral smear ?
• A peripheral blood smear or blood film is a thin layer of blood smeared on a microscope glass slide and then
stained in such a way to allow the various blood cells to be examined microscopically.
4. WEDGE BLOOD SMEAR
Specimen:
• Peripheral blood smear made from EDTA-anticoagulated blood.
• Smears should be made within 1 hour of blood collection from
EDTA specimens stored at room temperature to avoid distortion of
cell morphology
• Blood smears can also be made from finger prick blood directly onto
slide.
Equipment
• Spreaders
• Grease free washed glass slides
• Blood capillary tube or micropipette 10 µL
5. SMEAR PREPARATION
1. Place a drop of blood, about 2-3 mm
in diameter approximately 1 cm from
one end of slide.
2. Place the slide on a flat surface, and
hold the other end between your left
thumb and forefinger.
3. With your right hand, place the
smooth clean edge of a second
(spreader) slide on the specimen slide,
just in front of the blood drop.
4. Hold the spreader slide at a 30°- 45
angle, and draw it back against the
drop of blood
6. Allow the blood to spread almost to
the edges of the slide.
7. Push the spread forward with one
light, smooth moderate speed. A thin
film of blood in the shape of tongue.
8. Label one edge with patient name, lab
id and date.
9. The slides should be rapidly air dried by
waving the slides or using an electrical fan.
7. Characteristics of A Good Smear
1. Good smear is tongue shaped with a smooth tail.
2. Does not cover the entire area of the slide.
3. Has both thick and thin areas with gradual transition.
4. Does not contain any lines or holes.
8.
9. The thickness of the smear
Is determined by:
1. The angle of the spreader slide. (the greater the angle, the
thicker and shorter the smear).
2. Size of the blood drop.
3. Speed of spreading
i. If the hematocrit is increased, the angle of the spreader slide should be decreased.
ii. If the hematocrit is decreased, the angle of the spreader slide should be increased.
10. Common causes of a poor blood smear
1. Drop of blood too large or too small.
2. Spreader slide pushed across the slide in a jerky manner.
3. Failure to keep the entire edge of the spreader slide against the slide
while making the smear.
4. Failure to keep the spreader slide at a 30° angle with the slide
5. Failure to push the spreader slide completely across the slide.
6. Irregular spread with ridges and long tail: Edge of spreader dirty or
chipped; dusty slide
7. Holes in film: Slide contaminated with fat or grease and air bubbles.
8. Cellular degenerative changes: Delay in fixing, inadequate fixing time or
methanol contaminated with water.
12. Aim of blood smear
• Blood films are usually examined to investigate hematological
disorders and, occasionally, to look for parasites within the blood
such as malaria, filaria.
• Examination of thin blood films is important in the investigation
and management of anemia, 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 cost, useful
information about a patient’s condition.
14. Brief History
• The beginnings of modern-day blood staining can be traced back to Ehrlich,
who in 1877, was the first scientist to divide the aniline dyes into acidic and
basic categories.
• Three years later , the malarial parasite was discovered, and a rigorous search
for an improved blood stain ensued – In 1888, Chenzinsky discovered a stain
composed of cationic dye methylene blue and anionic eosin.
• Malachowski modified that stain with a markedly improved color range
and depth
• Regrettably, when he published his results in Berlin in 1891, he did not give the
necessary details as to how his method might be reproduced.
• 3 weeks later the young Russian protozoologist Romanowsky published similar
findings in an article so celebrated that all future stains of analogous
composition were named as Romanowsky-type stains
[Romanowsky noted that a neglected, moldy methylene blue solution was more effective
in producing the desired red plasmodial chromatin bodies than was a fresh solution]
19. • Wright’s Stain
• Components
• Stock solution:
• Wright’s stain 0.25gm to 0.5 gm
• Absolute acetone free methanol 100 ml
• Buffer (pH 6.8):
• KH2PO4 6.63 g
• Na2HPO4 2.56 g
• Distilled water 1,000 ml
• May–Grunwald Stain – 0.3 gm in 100 ml methanol
• Jenner – 0.5 gm in 100 ml methanol ( no over night incubation)
• Leishman’s Stain – 0.2 gm in 100 ml methanol
• Giemsa – 1 gm in 100 ml methanol
20. After drying
the smear
After drying the smear is
flooded with Wright stain
solution for 3 to 5 minutes,
Staining does not occur at
this stage
Buffer water of pH 6.8 is added
on the smear already
containing the stain solution.
(No washing)
Mixing of stain solution and
buffer water is done by blowing
in a figure of ‘8’ manner and kept
for 10 to 12 minutes for the
actual staining
Thoroughly washed under
slow running tap water.
Air drying and
examined under
light microscope
using a
temporary
mounting media
Wright staining followed in our Lab
21. Fixation is done
by immersing in
a jar of
methanol for 5–
10 min
Fixed smears are transferred to
a staining jar containing
May–Grunwald stain freshly
diluted with an equal volume
of buffered water and allowed
to stain for about 15 min.
Transferred
to a jar containing Giemsa’s
stain freshly diluted with 9
volumes of buffered water
without washing
After staining for 10–15 min,
slides are transferred to a jar
containing buffered water, pH
6.8
Rapid washing in 3 to 4
changes of water, and allow to
stand undisturbed
in water for a 2–5 minutes
Air drying and
examined under
light microscope
using a
temporary
mounting media
May–Grunwald–Giemsa Stain
22. Colour responses of blood cells to Romanowsky staining
• Cellular component
• Nuclei
• Nucleoli
• Chromatin
• Cytoplasm
• Erythroblast
• Erythrocyte
• Reticulocyte
Colour
Purple
Light blue
Dark blue
Dark pink
Grey–blue
24. Granules
• Promyelocyte(primary granules) Red or purple
• Basophil
• Eosinophil
• Neutrophil
• Toxic granules
• Platelet
Purple black
Red–orange
Purple
Dark blue
Purple
25. Other inclusions
• Auer body
• Cabot ring
• Howell-Jolly body
• Döhle body
Purple
Purple
Purple
Light blue
26. Factors influence smear staining method
• Blood smear may be under or over stained based
on the following
• Concentration of the stain used
– Low concentration: pale coloured cells (under
staining)
– High concentration: dark stained smear (over stained)
• Time of exposure the stain and the buffer
– Too long: overstaining,
– Too short: understaining
27. Factors giving rise to faulty staining
• Appearances
❖Too blue
Causes
Eosin concentration too low
Incorrect preparation of
stock stock stain exposed to
bright
daylight
Batch of stain solution overused
• Impure dyes
• Staining time too short
• Staining solution too acid
• Smear too thick
• Inadequate time in
buffer solution
28. Too pink Incorrect proportion of azure
B-eosin Y
• Impure dyes
• Buffer pH too low
• Excessive washing in buffer
solution
29. • Pale staining • Old staining solution
• Overused staining solution
• Incorrect preparation of
stock
• Impure dyes, especially
azure A and/or C
• High ambient temperature
30. •Neutrophil granules
not stained
•Neutrophil granules Dark
Blue/black (pseudo-toxic)
Insufficient azure B
Excess azure B
• Other stain anomalies Various
contaminating
dyes and metal salts
31. • Stain deposit
on film
Stain solution left in
uncovered jar
Stain solution not filtered
Inadequate fixation or
prolonged storage before
fixation
• Blood collected into heparin
as anticoagulant
• Blue background
32. Precautions
• PBS should be fixed as soon as possible after drying; background of
dried plasma may stain pale blue.
• Smear should not come in contact with water before fixation is
complete.
• Methanol must be stored in a bottle with a tightly fitting stopper
and not left exposed to the atmosphere, especially in humid
climates.
34. • Patient identification should be checked and confirmed and the microscope
slide matched with the corresponding CBC report.
• The film should be examined macroscopically to confirm adequate spreading
and to look for any unusual spreading or staining characteristics.
35. • PBS should be compared with the CBC report and a judgement
made as to whether the WBC, hemoglobin concentration (Hb), MCV
and platelet count are consistent with the smear.
(i) a poorly mixed or partly clotted or
hemolyzed specimen;
(ii) a specimen that is too small so that the
instrument has aspirated an inadequate volume;
(iii) the blood film and CBC being derived from
different blood specimens.
36. • On placing a pbs under the microscope, the first decision to be made is whether
or not it is suitable for further examination. Spreading, fixation and staining
must be satisfactory and there should be no artefactual changes produced by
excess EDTA or prolonged storage, e.g., spherocytosis, crenation or echinocytic
changes in red cells, degeneration of neutrophils and lobulation of some
lymphocyte nuclei
• RBCs are just touching but not overlapping.
• WBCs should be distributed regularly without undue concentration along the edges or
in the tail (can be seen in a very thin smear)
38. Red cell Morphology
• Terms used to describe a normal red cell morphology
• normocytic, which means that the cells are of normal size
• normochromic, which means that the cells have the normal concentration
of hemoglobin (i.e., when central pallor occupies ≤ one‐third of the red cell
diameter portion)
40. • Anisocytosis
• Anisocytosis is an increase in the variability of erythrocyte size beyond that which is
observed in a normal healthy subject.
41. • Dimorphic picture
• Indicates the presence of two distinct populations of red cells.
• Often applied when there is one population of hypochromic, microcytic cells and another
population of normochromic cells, either normocytic or macrocytic.
42. Chromasia
1. Hypochromia
➢ When central pallor is more than 1/3 of the red cell diameter
➢ Usually associated with microcytosis
(question – 1. Name 2 conditions where this association is not seen?
(thal trait , microspherocytosis, liver disease)
2. What are leptocytes ?)
2. Hyperchromia
➢ The term ‘hyperchromia’ is rarely used in describing smears.
➢ Deep staining of the red cells with a lack of central pallor
➢ May be seen in two circumstances: first, in the presence of macrocytes and second,
when cells are abnormally rounded
43. Leptocytes
• Thin red cells with large
unstained central area.
• Seen in
– Severe iron deficiency
anemia
– Thaleasaemia
44. 3. Anisochromasia
• Anisochromasia describes an increased variability in the degree of staining or haemoglobinisation of the red
cell
• In practice, it usually means that there is a spectrum of staining from hypochromic to normochromic.
• Indicates a changing situation, such as iron deficiency developing or responding to treatment or anaemia of
chronic disease developing or regressing.
45. 4. Polychromasia
• Polychromasia or polychromatophilia describes red cells that are pinkish-blue as a consequence of uptake
both of eosin, by hemoglobin, and of basic dyes, by residual nucleic acids.
• considerably larger than mature erythrocytes and, as a consequence of a reduced hemoglobin
concentration, are less dense
• have an irregular, multilobate surface
46. • Q. What is the normal percentage of polychromatophils in PBS?
48. Elliptocytes/ ovalocytes
• Both are oval in shape.
• A cell with a long axis
more than twice its short
axis should be designated
an elliptocyte
• while a cell with the long
axis less than twice its
short axis is designated
an ovalocyte
49. Spherocytes
• Nearly spherical
• Diameter is smaller than
normal
• Lack central pale area or
have a smaller , eccentric,
pale area
• Seen in
– hereditary spherocytosis
– Some cases of
autoimmune hemolytic
anemia
– direct physical or
chemical injury
50. Target cells
• Target cells have an area of increased staining, which appears in the
middle of the area of central pallor
• Formed as a consequence of a redundant membrane in relation to the
volume of the cytoplasm.
• They may also be thinner than normal cells.
• In vivo they are bell-shaped as demonstrated on scanning electron
microscopy.
• Flatten on spreading to form the characteristic target cell seen on light
microscopy
52. Echinocytes
• Also called crenated cells
• Erythrocytes that have lost their disc
shape and are covered with 10–30
short blunt spicules of fairly regular
form
• Commonly occur as an artifact during
preparation of film Hyperosmolarity
• Discocyte–echinocyte
transformation
• Overnight stored blood at 20 C
before films are made.
• Premature infant after
exchange transfusion
• water contaminating the
Wright’s stain (or absolute
methanol)
53. • Q – What is the usual mechanism of echinocyte formation in an
otherwise normal blood sample?
55. Acanthocytes
• Acanthocytes are cells of
approximately spherical shape
bearing between 2 to 20 spicules
that are of unequal length and
distributed irregularly over the red
cell surface.
• Probably results from a
preferential expansion of the
outer leaflet of the lipid bilayer.
• Seen in Abnormal phospholipid metabolism
• Abetalipoproteinemia
• Inherited abnormalities of red cell membrane protein
• Splenectomy
56. Keratocytes
• Have pairs of spicules;
usually single pair.
• Sometimes termed as
Bite cell or helmet cell
• Seen in
– Mechanical damage
– Removal of Heinz body by
pitting action of spleen.
57. Schistocytes
• These are fragmaented
erythrocytes.
• According to ICSH these
are red cell fragments with
sharp angles and straight
borders, small crescents,
helmet cells, keratocytes
and microspherocytes
• Quantification is per 1000
erythrocytes, with more
than 1% schistocytes being
regarded as significant
58. Stomatocytes
• Red cells with central biconcave
area appears slit like in dried film.
• Wet film it appears as cup-
shaped.
• Seen in
– Artifact
– south-east Asian
ovalocytosis
– liver disease,
– alcoholism,
– myelodysplastic
syndromes.
59. Sickle cell
• Cells are sickle (boat
shape) or crescent
shape
• Present in film of
patient with
homozygosity for Hb S.
• Usually absent in
neonates and rare in
patients with high Hb F
percentage
60. Teardropcells
• One side of cells is
tapered and other is
blunt.
• Usually found when
there is marrow failure
or fibrosis
61.
62. Red blood cell inclusions
Name of Inclusion
• Howell-Jolly body
• Basophilic stippling
• Pappenheimer body
• Heinz body(supravital only)
• Crystals
• Cabot rings
• Nucleus
• Content
• DNA
• RNA
• Iron
• Denatured hemoglobin
• Hemoglobin-C
• Mitotic spindle
remnants
• DNA
63. Basophilic Stippling
• Presence of irregular basophilic
granules with in Rbc which are
variable in size .
• Stain deep blue with Wright’s stain
• Fine stippling seen with
– Increased polychromatophilia
– Increased production of red cells.
• Coarse stippling
– Lead and heavy metal poisoning
– Disturbed erythropoiesis
• Megaloblastic anemia
• Thalassaemia
• infection
• liver disease
– Unstable Hb
– Pyrimidine-5’-nucleotidase def.
64. Howell-Jolly Bodies
• Smooth single large round
inclusions which are remnant
of nuclear chromatin.
• Seen in
• Single –
– Megaloblastic anemia
– Hemolytic anemia
– Postsplenectomy
• MULTIPLE –
– Megaloblastic anemia
– Abnormal erythropoiesis • Howell-Jolly Bodies
65. Pappenheimer Bodies
• These are small single or
multiple peripherally sited
angular basophilic (almost
black) erythrocyte inclusions.
• Smaller than Howell–Jolly
bodies.
• composed of haemosiderin.
• Their nature can be
confirmed by Perls’ stain.
• Seen in
– Sideroblastic
erythropoiesis
– Hypospenism
– Myelodysplastic syndrome
– Hemolytic anemia
66. Heinz bodies
• Seen on supravital stains
• Not seen on Romanowsky stain.
• Purple, blue, large, single or
multiple inclusions attached to the
inner surface of the red blood cell.
• Represent precipitated normal or
unstable hemoglobins.
• seen – Postsplenectomy
• Oxidative stress
– Glucose-6-phosphate
dehydrogenase deficiency,
– Glutathione synthetase
deficiency
– Drugs
– Toxins
– Unstable hemoglobins
67. Cabot Rings
• These are Ring shaped or figure of
eight or loop shaped.
• Red or Reddish purple with
Wright’s stain and have no internal
structure
• Observed rarely in
– Pernicious anemia,
– Lead poisoning,
68. Malarial stippling
• Fine granules of
plasmodium vivax
• On wright stain these
are fine , purplish red
• Red cells are larger than
normal
69. Rouleaux Formation
• Alignment of red cells one
upon another so that they
resemble stacks of coins.
• Occurs inParaproteinemia
( monoclonal
gammopathy)
• Elevated plasma
fibrinogen or globulin
level
70. Agglutination
• It is more irregular and
round clumping than
linear rouleaux
• Seen with cold
agglutinin
• Anti RBC antibody
• Autoimmune hemolytic
anemia
• Macroglobulinemia
71. • Discrepancies in Red cell parameters –
Falsely high RBC –
a) Numerous large platelets
b) Hyperlipidaemia
c) Cryoglobulinemia/
Cryofibrinogenemia
Falsely Low RBC –
a) Cold agglutinins (Rarely warm
autoantibodies)
b) EDTA‐dependent pan-
agglutination
c) In vitro red cell lysis due to
mishandling of Extreme
microcytosis or fragmentation
Falsely high MCV –
a) Storage of blood at room
temperature
b) Cold agglutinins and EDTA‐
dependent pan-agglutinins
c) very high WBC count
d) Hyperosmolar states (e.g.
hypernatraemia, )
e) Excess K2EDTA
Falsely Low MCV –
a) Increase in ambient
temperature
b) Hypo‐osmolar states (e.g.
hyponatraemia
c) Repeated mixing of sample
leading to increased
oxygenation
Falsely high RBC
a) Poorly mixed specimen
b) High WBC
c) Hyperlipidaemia,
[endogenous or due to
parenteral nutrition]
d) hypergammaglobuline
mia
73. ▪ Before evaluating leucocyte following must be seen-
Film is well made
Distribution of cells is uniform
Staining is satisfactory
▪ While scanning estimate the total leucocyte count
▪ Differential count is done at oil immersion
74. • Ten microscopic fields are examined in a vertical direction from bottom to
top or top to bottom
• Slide is horizontally moved to the next field
• Ten microscopic fields are counted verticallyinoppositetothe previousdirection.
• Procedure is repeated until 100 WBCS have been counted (zig zag motion)
Scanning technique for WBC differential count and
morphologic evaluation
75. Manual differential counts
• These counts are done in the same area as
WBC and platelet estimates with the red cells
barely touching.
• Count 100 WBCs under 40x/100x including
all cell lines from immature to mature.
✓Reporting results
• Approximate absolute number of cells/µl = %
of cell type in differential x white cell count
76. •If 10 or more nucleated RBC's (NRBC) are
seen, correct the
White Count using this formula:
Corrected WBC Count =
WBC x 100/( NRBC + 100)
Example : If WBC = 5000 and 10 NRBCs have
been counted
Then we have actually counted 110 cells among which
100 cells are WBCs.
That means in 1 cell there will 100/110 WBC.
The corrected white count is (100/110) x 5000 = 4545.50
77. Leukocytes Normally Present in Blood
Neutrophils ( polymorphonuclear leucocytes)
Band form
Eosinophils
Basophils
Lymphocytes
Monocytes
78. POLYMORPHONUCLEAR NEUTROPHILS
• 40 to 80 percent of total
WBC count(2.0–7.0 ×109/l )
• Diameter - 13 µm
• segmented nucleus and
pink/orange cytoplasm with
fine granulation(0.2-0.3µm)
stain tan to pink with
Wright’s
• Lobes -2-5
• small percent has four lobes
and occasionally five lobes.
79. Band forms
• neutrophils has either a
strand of nuclear material
thicker than a filament
connecting the lobes, or a U-
shaped nucleus of uniform
thickness.
• Up to 8% of circulating
neutrophils are
unsegmented or
partly segmented (‘band’
forms)
81. Myeloblast (0%)
Promyelocyte (0%)
Myelocyte (<0.5%)
Metamyelocyte (<1%)
Band form (5 to 8%)
Bilobed neutrophils (30-35%)
Trilobed neutrophils (40-50%)
Tetra lobed neutrophils (15-20%)
Penta lobed (<3%)
Hexalobed (0%)
Left Shift
Right Shift
82. Granules
• Toxic granulation-
increase in staining
density and number of
granules
• Seen with Bacterial
infections and other
inflammation
• Administration of G-CSF
• Anaplastic anemia
84. Alder–Reilly anomaly
• Granules are large,
– discrete,
– stain deep red
– may obscure the
nucleus
– Neutrophil function
is Normal
85. Chédiak-Higashi Syndrome
• Granules are
– also seen in other
leukocytes like
lymphocytes
– Giant
– Scanty azurophilic
– functional defect occur
86. Dohle Bodies
• Small, round or oval, pale
blue-grey structure
• Found at periphery of
neutrophil.
• Contains Ribosomes and
Endoplasmic reticulum
• Seen in – Bacterial infection
– inflammation
– administration of G-CSF
– during pregnancy
87. May–Hegglin anomaly
• inclusions occur in all
types of leucocytes
except lymphocytes.
• contain small basophilic
cytoplasmic granules
88. Vacuoles in neutrophils
• In Fresh blood smear
• vacuoles seen in
– severe sepsis
– as an artifact with
prolonged standing
89. Nuclei
• Hypersegmentated
neutrophil
• def.-presence of neutrophils
with six or more lobes or the
presence of more
than 3% of neutrophils
with at least five lobes.
• seen in Megaloblastic
anemia
– uraemia
– iron deficiency.
– Drugs-cytotoxic treatment with
Methotrexate
– hydroxycarbamide
90. Pelger–Huët Cells
• Pelger–Huët anomaly
• Benign inherited
condition.
• Neutrophil nuclei fail to
segment properly.
• Majority of circulating
neutrophils have only
two discrete equal-sized
lobes connected by a
thin chromatin bridge.
91. Pseudo-Pelger cells
• Pseudo-Pelger cells or the
acquired Pelger– Huët
anomaly
• Acquired condition
• Morphologically similar to
Pelger–Huët anomaly
• seen in Myelodysplastic
syndromes,
• Acute myeloid leukaemia
with dysplastic maturation,
• Occasionally in chronic
myelogenous leukaemia
92. Pyknotic Neutrophils (Apoptosis)
• Small numbers of dead
or dying cells may
normally be found in
the blood
• seen in infections
– invitro after standing for
12-18 hrs
– Nuclei-round dense,
featureless
– Cytoplasm-dark pink
95. • Eosinopenia- seen with prolonged steroid administration.
• Eosinophilia-
asthma
Allergic
conditions
• severe eosinophilia-
- parasitic infection
– reactive eosinophilia
– Eosinophilic leukaemia
– Idiopathic hypereosinophilic syndrome
– T-cell lymphoma, B-cell lymphoma
and acute lymphoblastic leukaemia.
96. BASOPHILS• Rarest <1%
• Nucleus segments fold up
on each other resulting
compact irregular dense
nucleus(closed lotus flower
like)
• Granules-large, variable size
dark blue or purple often
obscure the nucleus
• Granules are rich in
histamine, serotonin and
heparin
• Increase in
myeloproliferative disorder-
CML
97. MONOCYTES
• 2-10% of total wbc count
• Size- largest circulating leucocyte, 15–18µm
in diameter
• Cytoplasm- grey blue, may be vacuolated
• Nucleus- large , curved , horse shoe
shape/kidney/ fetal shaped
• No segmentation occur
• Chromatin- fine evenly distributed
• Increase in chronic infections and
inflammatory conditions such as
– TuberculosisandCrohn’sdisease,
– Chronic myeloid leukaemias
– Acute leukaemias with a
monocytic component
– Infectious mononucleosis
98. LYMPHOCYTES
• 20-40% of total wbc count
• two types
1. Small lymphocyte(6-10µm)
2. Large lymphocyte(12-15µm)
• Nucleus-single, sharply
defined, stain dark blue on
Wright’s stain
• Cytoplasm- Pale blue
• Large lymphocytes less
densely stain nuclei &
abundant cytoplasm
• Few round purple(azure)
granules are present
99. Turk cells
• Türk’ cell (immunoblasts)-
Transformed lymphocyte seen
in bacterial and viral infection
• Size 10-15 µm
• Nucleus- Round,
• Large nucleolus, and abundant,
deeply basophilic cytoplasm
100. Reactive lymphocytes
• Have slightly larger
nuclei with more
open chromatin
• Abundant cytoplasm
that may be
irregular.
• Seen in -infectious
mononucleosis
– viral infections
101. What are Downey cells
Type I: Small cells with minimum cytoplasm, indented
nucleus/irregular nuclear membrane, and condensed chromatin.
▪Type II: Larger cells with abundant cytoplasm; the lymphocyte
cytoplasm seems to hug the red cells. Type II is the most common
type of Downey cell.
▪Type III: Cells with large moderate basophilic cytoplasm and nucleus
with coarse chromatin. Nucleoli are apparent
104. Leucoerythroblastic blood films
• A blood film is referred to as leucoerythroblastic if it contains NRBC
and granulocyte precursors.
105. • Discrepancies in WBC parameters –
Falsely high TLC–
a) Presence of NRBCs
b) Liver disease
c) Cold agglutinins
d) Numerous giant platelets or megakaryocyte
fragments
e) Platelet aggregates
f) Cryoglobulinemia/ Cryofibrinogenemia
g) Malarial parasites
h) Systemic mycoses
i) abnormal haemoglobins (in some counters)
Falsely Low RBC –
a) Cell lysis caused when blood is more than 3
days old
b) Storage at room temperature for 24 hours or
more
c) Storage at 4°C for more than 24 hours
(Horiba)
d) Leucocyte or leucocyte and platelet
aggregation due to an antibody or to
alteration of the cell membrane or to the
presence of neoplastic cells
e) In vitro red cell lysis due to mishandling of
Extreme microcytosis or fragmentation
106. Platelates
• Size -1-3µm
• Normal count - 280 ±130×109/µl
• Non nucleated cells derived from cytoplasmic
fragments of Megakaryocytes
• Has purple red granules.
• Liliac color
110. Platelet morphology: Giant platelets
• Platelates seems to be
size of rbcs.
• Seen in
– May –Hegglin anomly
– Bernard Soulier
syndrome
– Alport syndrome
– Storage pool syndrome
113. • 10 oil immersion fields counting
• Average
• Multiply by 15000.
114. • Discrepancies in Platelet parameters –
Falsely Low platelet count–
a) Partial clotting of specimen
b) Activation of platelets during venepuncture with
consequent aggregation
c) Activation of platelets during cardiopulmonary bypass
d) EDTA‐induced platelet aggregation (more common in viral
infections, particularly hepatitis A, CMV and inluenza A)
e) EDTA‐induced platelet degranulation and swelling
f) Lipiodol‐induced platelet clumping following
chemoembolisation
g) Platelet satellitism
h) Platelet phagocytosis by neutrophils and monocytes
i) Storage of blood at 4°C for more than 24 hours
j) Giant platelets falling above upper threshold for
platelet count
k) Heparin addition to blood sample in patient with
antibodies to heparin‐platelet factor 4
Falsely High platelet count –
a) Microcytic red cells or red cell fragments failing
below upper threshold for the platelet count
b) Presence of Microspherocytes (in hereditary
spherocytosis, burns)
c) Inadvertent heating of blood sample
d) White cell fragments counted as platelets
(fragments of leukaemic blast cells, hairy cells
or lymphoma cells)
e) Haemoglobin H disease
f) Hypertriglyceridaemia or hyperlipidaemia
115. changes on prolonged storage of sample
• EFFECT ON COUNT-
• Less marked in blood in ACD, CPD or Alsever’s solution
than in EDTA.
• At room temperature blood is stable up to 8 h.
• RBC
– Swell up the PCV and MCV increases
– Osmotic fragility increases
– Erythrocyte sedimentation rate decreases
– At 4ͦ C up to 24 h
– Reticulocyte count- Unchanged upto 24 h at 4 C
– Hemoglobin Unchanged upto 2-3 days
116. Disadvantages of the Peripheral Blood
Smear
• Experience is required to make technically
adequate smears.
• Non-uniform distribution of white blood cells
• Larger leukocytes concentrated near edges and
lymphocytes scattered throughout.
• Non-uniform distribution of RBCs
– Small crowded red blood cells at the thick edge
– Large flat red blood cells without central pallor at the
feathered edge
117. EXAMINATION OF BLOOD FILMS FOR
PARASITES
• thick film- when parasites are scanty
• thin film – identification of species
• STAINING OF FILM
– by Leishman’s stain at pH 7.2
120. Schizonts are commonly seen in P. vivax infection and appear
as large bodies containing 12 to 24 nuclei and a loose
pigmented body. This photograph shows an early schizont of P.
vivax on the left and mature schizonts