Bone marrow examination is an indispensable tool for diagnosing blood disorders. It involves bone marrow aspiration and biopsy. Aspiration provides cytological details for diagnosis while biopsy assesses cellularity and infiltration. Together they provide important information for evaluating diseases of the blood. The procedure involves inserting needles into bone (typically the iliac crest) under local anesthesia to extract bone marrow samples. Aspirates are used to make smears for staining and analysis under the microscope. Biopsies provide tissue samples for histological examination after fixation and processing. Both tools are used to evaluate bone marrow cellularity, identify abnormal cells, and make diagnoses of blood disorders and other conditions.
The Coombs test, also known as the direct antiglobulin test (DAT) or indirect antiglobulin test (IAT), detects antibodies or complement coating red blood cells. It involves sensitizing RBCs with patient serum, washing unbound antibodies, then adding anti-human globulin reagent to form a "bridge" and cause agglutination if antibodies or complement are present on the RBCs. Controls like Coombs control check cells are used to validate negative results and detect technical problems. The DAT detects in vivo coating while the IAT detects in vitro coating during antibody screening and identification.
The document discusses the history, utility, and methods of preparing cell blocks from fine needle aspiration cytology samples. Cell blocks allow examination of histological structure and use of ancillary tests. Key methods include the fixed sedimentation method using a 1:1 ratio of 100% alcohol and 40% formalin, the plasma thrombin method using equal parts plasma and thrombin, and the bacterial agar method using 3% agar. Cell blocks provide increased diagnostic sensitivity and specificity compared to cytology alone through examination of tissue architecture and ability to perform special stains and molecular testing.
This presentation discusses the cytology of cerebrospinal fluid (CSF). CSF is normally clear, colorless, and transparent. It is produced in the cerebral ventricles and circulates in the spinal canal. A lumbar puncture or spinal tap is performed to examine the physical appearance and microscopic evaluation of CSF slides. The cytology technique involves centrifuging CSF samples onto slides, staining with Papanicolaou stain, and examining under a microscope. Normal CSF is acellular, while abnormal CSF may contain lymphocytes, monocytes, malignant or leukemia cells, indicating conditions such as meningitis, subarachnoid hemorrhage, or CNS malignancy. Cytology of CSF aids in the diagnosis of various
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 summarizes different types of erythrocyte inclusions seen on supravital and Wright staining, along with their composition and associated disease states. It describes inclusions such as basophilic stippling composed of RNA seen in iron deficiency, Howell-Jolly bodies which are nuclear fragments seen in megaloblastic anemia, Heinz bodies containing denatured hemoglobin associated with G6PD deficiency, and siderotic granules containing iron found in sideroblastic anemia.
An excellent ppt on basics of bone marrow morphology and examination which i came accross on the internet.. Not my creation.. Full credit to the author..
Sputum examination provides important diagnostic information by analyzing material coughed up from the lungs and respiratory tract. Key indications for sputum examination include identifying the causative organism in suspected lower respiratory infections like pneumonia or tuberculosis. Sputum samples can also be examined cytologically to detect malignant cells or investigate other infections. Proper collection and transport of sputum samples is important for microbiological culture and other tests. Staining and microscopic examination of sputum looks for bacteria, fungi, parasites and other pathogenic organisms. Molecular tests like PCR provide a rapid and sensitive method for tuberculosis diagnosis.
This document discusses leukocyte disorders, which can be either malignant (tumors) or non-malignant. It focuses on non-malignant disorders affecting neutrophils. Neutrophils and other leukocytes are produced through hematopoiesis in the bone marrow from stem cells. The document describes the different types of leukocytes (neutrophils, eosinophils, basophils, monocytes, lymphocytes), their functions, and normal ranges. It also discusses disorders characterized by increased or decreased levels of these cells.
The Coombs test, also known as the direct antiglobulin test (DAT) or indirect antiglobulin test (IAT), detects antibodies or complement coating red blood cells. It involves sensitizing RBCs with patient serum, washing unbound antibodies, then adding anti-human globulin reagent to form a "bridge" and cause agglutination if antibodies or complement are present on the RBCs. Controls like Coombs control check cells are used to validate negative results and detect technical problems. The DAT detects in vivo coating while the IAT detects in vitro coating during antibody screening and identification.
The document discusses the history, utility, and methods of preparing cell blocks from fine needle aspiration cytology samples. Cell blocks allow examination of histological structure and use of ancillary tests. Key methods include the fixed sedimentation method using a 1:1 ratio of 100% alcohol and 40% formalin, the plasma thrombin method using equal parts plasma and thrombin, and the bacterial agar method using 3% agar. Cell blocks provide increased diagnostic sensitivity and specificity compared to cytology alone through examination of tissue architecture and ability to perform special stains and molecular testing.
This presentation discusses the cytology of cerebrospinal fluid (CSF). CSF is normally clear, colorless, and transparent. It is produced in the cerebral ventricles and circulates in the spinal canal. A lumbar puncture or spinal tap is performed to examine the physical appearance and microscopic evaluation of CSF slides. The cytology technique involves centrifuging CSF samples onto slides, staining with Papanicolaou stain, and examining under a microscope. Normal CSF is acellular, while abnormal CSF may contain lymphocytes, monocytes, malignant or leukemia cells, indicating conditions such as meningitis, subarachnoid hemorrhage, or CNS malignancy. Cytology of CSF aids in the diagnosis of various
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 summarizes different types of erythrocyte inclusions seen on supravital and Wright staining, along with their composition and associated disease states. It describes inclusions such as basophilic stippling composed of RNA seen in iron deficiency, Howell-Jolly bodies which are nuclear fragments seen in megaloblastic anemia, Heinz bodies containing denatured hemoglobin associated with G6PD deficiency, and siderotic granules containing iron found in sideroblastic anemia.
An excellent ppt on basics of bone marrow morphology and examination which i came accross on the internet.. Not my creation.. Full credit to the author..
Sputum examination provides important diagnostic information by analyzing material coughed up from the lungs and respiratory tract. Key indications for sputum examination include identifying the causative organism in suspected lower respiratory infections like pneumonia or tuberculosis. Sputum samples can also be examined cytologically to detect malignant cells or investigate other infections. Proper collection and transport of sputum samples is important for microbiological culture and other tests. Staining and microscopic examination of sputum looks for bacteria, fungi, parasites and other pathogenic organisms. Molecular tests like PCR provide a rapid and sensitive method for tuberculosis diagnosis.
This document discusses leukocyte disorders, which can be either malignant (tumors) or non-malignant. It focuses on non-malignant disorders affecting neutrophils. Neutrophils and other leukocytes are produced through hematopoiesis in the bone marrow from stem cells. The document describes the different types of leukocytes (neutrophils, eosinophils, basophils, monocytes, lymphocytes), their functions, and normal ranges. It also discusses disorders characterized by increased or decreased levels of these cells.
1. Cytology of body fluids involves examining fluids from various body cavities including cerebrospinal fluid, pleural fluid, peritoneal fluid, pericardial fluid, and synovial fluid. Specimen collection and laboratory analysis includes gross examination, cell counts, biochemical analysis, and microscopic examination.
2. Transudates and exudates are distinguished based on characteristics like protein content and cell differentials. Infection, inflammation, and malignancy can be identified by analyzing changes in fluid characteristics.
3. Cytology of body fluids provides diagnostic information for conditions affecting various organ systems. Proper collection and analysis of physical and chemical properties aids in differential diagnosis.
Paroxysmal Nocturnal Hemoglobinuria (PNH) is a rare, acquired hemolytic anemia caused by a loss of proteins that regulate the complement pathway, making red blood cells sensitive to complement-mediated lysis. It is characterized by intravascular hemolysis that is worse at night due to relative acidosis during sleep. Affected cells lack proteins like CD59 and CD55 on their surface. Diagnosis involves flow cytometry to detect absent or decreased CD59 and CD55.
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.
Immunophenotyping is a technique used to study the proteins expressed by cells using antibodies conjugated to fluorescent dyes. This can be done using flow cytometry to analyze cell suspensions, or immunohistochemistry (IHC) to study cells in tissue sections. Flow cytometry works by passing single cells through a laser which excites fluorescent markers, while IHC visualizes the location of antigen-antibody complexes in tissue. Both techniques are useful for research, diagnosis, and identifying cell lineages through specific protein markers.
Reticulocytes are immature red blood cells that are released from the bone marrow into circulation during erythropoiesis. Automated analysis of reticulocytes provides several parameters that can help evaluate bone marrow function and iron status. The immature reticulocyte fraction is useful for monitoring bone marrow recovery after chemotherapy or transplant. Reticulocyte hemoglobin content measures like CHr can detect iron deficiency earlier than other tests. Using multiple reticulocyte parameters in diagnostic algorithms helps classify anemias and guide treatment.
Immunofluorescence is a technique that allows visualization of specific proteins or antigens in tissue sections. It involves binding antibodies conjugated to fluorescent dyes. There are two main types - direct immunofluorescence detects in vivo antibodies bound to tissue, while indirect detects circulating antibodies in patient serum. Immunofluorescence microscopy uses filters and dichroic mirrors to generate fluorescent images. It has various applications in histopathology for diseases like renal biopsy, skin biopsy, and more. Specific patterns of immunofluorescence staining can help diagnose glomerular diseases like post-infectious glomerulonephritis, membranoproliferative glomerulonephritis, and IgA nephropathy.
This document discusses white blood cell disorders, focusing on quantitative disorders of the white blood cells. It describes leukocytosis, which is an increased number of white blood cells, and leukopenia, which is a decreased number. The main types of leukocytosis and leukopenia discussed are neutrophilic leukocytosis, lymphocytosis, and eosinophilia. The causes, pathophysiology, and clinical features of each of these conditions are explained in detail.
The Paris System for Reporting Urinary CytologyRawa Muhsin
The Paris System for Reporting Urinary Cytology provides standardized diagnostic categories for urine cytology specimens. It divides results into negative for high-grade urothelial carcinoma, positive for high-grade urothelial carcinoma, atypical urothelial cells, and suspicious for high-grade urothelial carcinoma based on the number and features of abnormal cells seen. The system aims to determine whether high-grade urothelial carcinoma is present or not, as this has important implications for patient management and prognosis. Risk of malignancy increases from negative to atypical to suspicious to positive categories.
Autoimmune hemolytic anemia (or autoimmune haemolytic anaemia; AIHA) occurs when antibodies directed against the person's own red blood cells (RBCs) cause them to burst (lyse), leading to insufficient plasma concentration.
This document discusses various platelet indices used in interpreting platelet histograms and their clinical significance. It defines plateletcrit (PCT) as the volume of platelets expressed as a percentage of total blood volume, with a normal range of 0.22-0.24%. It describes platelet distribution width (PDW) as measuring the variation in platelet size, with a normal range of 25-65, and higher values associated with vascular disease or cancer. It also defines platelet large cell ratio (P-LCR) as measuring larger platelets between 12-30 femtoliters, with increased levels seen in hyperlipidemia and autoimmune thrombocytopenic purpura. Mean platelet volume (MPV) is defined as the average
This document provides an overview of high-performance liquid chromatography (HPLC) and its use in pathology. It describes the basic components and separation mechanisms of chromatography, including mobile phase, stationary phase, and how separation occurs. It then discusses various types of chromatography techniques like ion exchange, partition, size exclusion, and affinity chromatography. The document focuses on HPLC, describing its instrumentation including reservoirs, pumps, injectors, columns, and detectors. It explains how HPLC provides improved resolution, speed, and reproducibility over other chromatography methods.
Diagnostic approach to pediatric malignant small round cellSivaranjini N
This document provides information on pediatric malignant small round cell tumors. It discusses key characteristics of these tumors including their appearance as relatively small, undifferentiated cells with a high nuclear-cytoplasmic ratio. Common small round cell tumors of childhood discussed include neuroblastoma, Wilms tumor, retinoblastoma, medulloblastoma, hepatoblastoma, and others. The document outlines approaches to classifying and diagnosing these tumors based on their origin, cytogenetics, immunohistochemistry profiles, and other features. Detailed information is provided on diagnostic criteria and clinical characteristics of neuroblastoma, Wilms tumor, and retinoblastoma.
This document provides information on chronic myelogenous leukemia (CML). It defines CML as an acquired myeloproliferative neoplasm caused by the presence of the Philadelphia chromosome and BCR-ABL fusion gene in the stem cells. CML typically affects adults aged 40-50 years and presents with excessive myeloid cells in the blood and bone marrow. The disease progresses through chronic, accelerated and blastic phases as the percentage of blasts increases and symptoms worsen over time if left untreated. Diagnosis involves blood tests showing elevated white cells, basophils and blasts as well as bone marrow biopsy demonstrating hypercellularity with myeloid hyperplasia.
CSF - Cerebrospinal fluid examination - from tapping to pathological diagnosisAshish Jawarkar
This is a series of notes on clinical pathology, useful for undergraduate and postgraduate students, as well as practising pathologists. Prepared from standard text books with data in tabular and easily readable format
Broncho-Alveolar Lavage Fluid Analysis provides information on the status of the respiratory tract beyond what can be seen bronchoscopically. It involves instilling saline into segments of the lung and analyzing the cell types in the returned fluid. A satisfactory sample contains at least 2x10^6 cells including over 10 macrophages per field. Differential cell counts can indicate conditions like pneumonia, cancer, sarcoidosis, and alveolar hemorrhage. Special stains can identify pathogens, lipids, proteins, and minerals for diagnostic purposes. Complications are generally minor but loss of lung function is a risk in severely compromised patients.
This document discusses Perls stain, which is used to identify iron deposits in tissue samples. It provides background on pigments in living tissue, including endogenous pigments like hemosiderin and hematogenous pigments. The history of Prussian blue and its use as Perls stain is described. The principle of the stain is that hydrochloric acid releases ferric ions from hemosiderin, which then react with potassium ferrocyanide to form insoluble Prussian blue pigment. Staining protocols, quality control, and clinical applications for identifying iron deposits in organs are covered.
CSF:
Derived through ultrafilteration and secretion through choroid plexus, produced at the rate of 500 ml/day.
Provides physical support, collects wastes, circulates nutrients and lubricates the CNS.
Normal CSF volumes:
In Adults: 90 - 150 ml
In Neonates: 10 - 60 ml
Total CSF volume is replaced every 5-7 hours.
COLLECTION
Lumbar puncture, Cisternal puncture, Lateral cervical puncture, Shunts and cannulas
Opening pressure – 90-180 mm H2O
Approximately 15-20 cc fluid collected
LAB
REQUIRED
Opening CSF pressure
Total cell count
Differential cell count
Glucose
Total protein
OPTIONAL
Cultures, Gram stain, AFB, Fungal and bacterial
antigens, Enzymes, PCR, Cytology, Electrophoresis,
VDRL, D-Dimers
This document discusses transfusion-transmitted infections (TTIs) and methods for screening donated blood. It notes that TTIs include viruses like HIV, HBV, HCV that can remain undetected in the blood donor but be transmissible. Screening methods include serological tests like ELISA, CLIA, rapid tests, as well as nucleic acid amplification tests (NAATs) that can detect infections earlier. Implementing individual donor NAT in addition to serological screening provides an additional safety layer and reduces the risk window period for TTIs in blood donations.
Bone marrow biopsy and aspiration provides qualitative and quantitative assessment of hematopoiesis. It involves extracting bone marrow samples via biopsy or aspiration, usually from the iliac crest, to examine for abnormalities. The bone marrow is examined microscopically via smears, crush preparations, or sections after staining. This allows evaluation of cellularity, maturation and morphology of cell lines, and detection of infiltrative processes or malignancies. Findings are essential for diagnosis of blood disorders and marrow failure.
This document provides information about bone marrow aspiration and biopsy procedures. It discusses the history and development of the procedures, anatomy of bone and bone marrow, indications for bone marrow examinations, how to perform aspirations and biopsies, complications, interpretation of results including cellularity, hematopoiesis, iron stores, and reticulin, and examples of bone marrow findings in different conditions like infections, deficiencies, and cancers. Bone marrow examinations provide important diagnostic information for hematological diseases and disorders.
1. Cytology of body fluids involves examining fluids from various body cavities including cerebrospinal fluid, pleural fluid, peritoneal fluid, pericardial fluid, and synovial fluid. Specimen collection and laboratory analysis includes gross examination, cell counts, biochemical analysis, and microscopic examination.
2. Transudates and exudates are distinguished based on characteristics like protein content and cell differentials. Infection, inflammation, and malignancy can be identified by analyzing changes in fluid characteristics.
3. Cytology of body fluids provides diagnostic information for conditions affecting various organ systems. Proper collection and analysis of physical and chemical properties aids in differential diagnosis.
Paroxysmal Nocturnal Hemoglobinuria (PNH) is a rare, acquired hemolytic anemia caused by a loss of proteins that regulate the complement pathway, making red blood cells sensitive to complement-mediated lysis. It is characterized by intravascular hemolysis that is worse at night due to relative acidosis during sleep. Affected cells lack proteins like CD59 and CD55 on their surface. Diagnosis involves flow cytometry to detect absent or decreased CD59 and CD55.
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.
Immunophenotyping is a technique used to study the proteins expressed by cells using antibodies conjugated to fluorescent dyes. This can be done using flow cytometry to analyze cell suspensions, or immunohistochemistry (IHC) to study cells in tissue sections. Flow cytometry works by passing single cells through a laser which excites fluorescent markers, while IHC visualizes the location of antigen-antibody complexes in tissue. Both techniques are useful for research, diagnosis, and identifying cell lineages through specific protein markers.
Reticulocytes are immature red blood cells that are released from the bone marrow into circulation during erythropoiesis. Automated analysis of reticulocytes provides several parameters that can help evaluate bone marrow function and iron status. The immature reticulocyte fraction is useful for monitoring bone marrow recovery after chemotherapy or transplant. Reticulocyte hemoglobin content measures like CHr can detect iron deficiency earlier than other tests. Using multiple reticulocyte parameters in diagnostic algorithms helps classify anemias and guide treatment.
Immunofluorescence is a technique that allows visualization of specific proteins or antigens in tissue sections. It involves binding antibodies conjugated to fluorescent dyes. There are two main types - direct immunofluorescence detects in vivo antibodies bound to tissue, while indirect detects circulating antibodies in patient serum. Immunofluorescence microscopy uses filters and dichroic mirrors to generate fluorescent images. It has various applications in histopathology for diseases like renal biopsy, skin biopsy, and more. Specific patterns of immunofluorescence staining can help diagnose glomerular diseases like post-infectious glomerulonephritis, membranoproliferative glomerulonephritis, and IgA nephropathy.
This document discusses white blood cell disorders, focusing on quantitative disorders of the white blood cells. It describes leukocytosis, which is an increased number of white blood cells, and leukopenia, which is a decreased number. The main types of leukocytosis and leukopenia discussed are neutrophilic leukocytosis, lymphocytosis, and eosinophilia. The causes, pathophysiology, and clinical features of each of these conditions are explained in detail.
The Paris System for Reporting Urinary CytologyRawa Muhsin
The Paris System for Reporting Urinary Cytology provides standardized diagnostic categories for urine cytology specimens. It divides results into negative for high-grade urothelial carcinoma, positive for high-grade urothelial carcinoma, atypical urothelial cells, and suspicious for high-grade urothelial carcinoma based on the number and features of abnormal cells seen. The system aims to determine whether high-grade urothelial carcinoma is present or not, as this has important implications for patient management and prognosis. Risk of malignancy increases from negative to atypical to suspicious to positive categories.
Autoimmune hemolytic anemia (or autoimmune haemolytic anaemia; AIHA) occurs when antibodies directed against the person's own red blood cells (RBCs) cause them to burst (lyse), leading to insufficient plasma concentration.
This document discusses various platelet indices used in interpreting platelet histograms and their clinical significance. It defines plateletcrit (PCT) as the volume of platelets expressed as a percentage of total blood volume, with a normal range of 0.22-0.24%. It describes platelet distribution width (PDW) as measuring the variation in platelet size, with a normal range of 25-65, and higher values associated with vascular disease or cancer. It also defines platelet large cell ratio (P-LCR) as measuring larger platelets between 12-30 femtoliters, with increased levels seen in hyperlipidemia and autoimmune thrombocytopenic purpura. Mean platelet volume (MPV) is defined as the average
This document provides an overview of high-performance liquid chromatography (HPLC) and its use in pathology. It describes the basic components and separation mechanisms of chromatography, including mobile phase, stationary phase, and how separation occurs. It then discusses various types of chromatography techniques like ion exchange, partition, size exclusion, and affinity chromatography. The document focuses on HPLC, describing its instrumentation including reservoirs, pumps, injectors, columns, and detectors. It explains how HPLC provides improved resolution, speed, and reproducibility over other chromatography methods.
Diagnostic approach to pediatric malignant small round cellSivaranjini N
This document provides information on pediatric malignant small round cell tumors. It discusses key characteristics of these tumors including their appearance as relatively small, undifferentiated cells with a high nuclear-cytoplasmic ratio. Common small round cell tumors of childhood discussed include neuroblastoma, Wilms tumor, retinoblastoma, medulloblastoma, hepatoblastoma, and others. The document outlines approaches to classifying and diagnosing these tumors based on their origin, cytogenetics, immunohistochemistry profiles, and other features. Detailed information is provided on diagnostic criteria and clinical characteristics of neuroblastoma, Wilms tumor, and retinoblastoma.
This document provides information on chronic myelogenous leukemia (CML). It defines CML as an acquired myeloproliferative neoplasm caused by the presence of the Philadelphia chromosome and BCR-ABL fusion gene in the stem cells. CML typically affects adults aged 40-50 years and presents with excessive myeloid cells in the blood and bone marrow. The disease progresses through chronic, accelerated and blastic phases as the percentage of blasts increases and symptoms worsen over time if left untreated. Diagnosis involves blood tests showing elevated white cells, basophils and blasts as well as bone marrow biopsy demonstrating hypercellularity with myeloid hyperplasia.
CSF - Cerebrospinal fluid examination - from tapping to pathological diagnosisAshish Jawarkar
This is a series of notes on clinical pathology, useful for undergraduate and postgraduate students, as well as practising pathologists. Prepared from standard text books with data in tabular and easily readable format
Broncho-Alveolar Lavage Fluid Analysis provides information on the status of the respiratory tract beyond what can be seen bronchoscopically. It involves instilling saline into segments of the lung and analyzing the cell types in the returned fluid. A satisfactory sample contains at least 2x10^6 cells including over 10 macrophages per field. Differential cell counts can indicate conditions like pneumonia, cancer, sarcoidosis, and alveolar hemorrhage. Special stains can identify pathogens, lipids, proteins, and minerals for diagnostic purposes. Complications are generally minor but loss of lung function is a risk in severely compromised patients.
This document discusses Perls stain, which is used to identify iron deposits in tissue samples. It provides background on pigments in living tissue, including endogenous pigments like hemosiderin and hematogenous pigments. The history of Prussian blue and its use as Perls stain is described. The principle of the stain is that hydrochloric acid releases ferric ions from hemosiderin, which then react with potassium ferrocyanide to form insoluble Prussian blue pigment. Staining protocols, quality control, and clinical applications for identifying iron deposits in organs are covered.
CSF:
Derived through ultrafilteration and secretion through choroid plexus, produced at the rate of 500 ml/day.
Provides physical support, collects wastes, circulates nutrients and lubricates the CNS.
Normal CSF volumes:
In Adults: 90 - 150 ml
In Neonates: 10 - 60 ml
Total CSF volume is replaced every 5-7 hours.
COLLECTION
Lumbar puncture, Cisternal puncture, Lateral cervical puncture, Shunts and cannulas
Opening pressure – 90-180 mm H2O
Approximately 15-20 cc fluid collected
LAB
REQUIRED
Opening CSF pressure
Total cell count
Differential cell count
Glucose
Total protein
OPTIONAL
Cultures, Gram stain, AFB, Fungal and bacterial
antigens, Enzymes, PCR, Cytology, Electrophoresis,
VDRL, D-Dimers
This document discusses transfusion-transmitted infections (TTIs) and methods for screening donated blood. It notes that TTIs include viruses like HIV, HBV, HCV that can remain undetected in the blood donor but be transmissible. Screening methods include serological tests like ELISA, CLIA, rapid tests, as well as nucleic acid amplification tests (NAATs) that can detect infections earlier. Implementing individual donor NAT in addition to serological screening provides an additional safety layer and reduces the risk window period for TTIs in blood donations.
Bone marrow biopsy and aspiration provides qualitative and quantitative assessment of hematopoiesis. It involves extracting bone marrow samples via biopsy or aspiration, usually from the iliac crest, to examine for abnormalities. The bone marrow is examined microscopically via smears, crush preparations, or sections after staining. This allows evaluation of cellularity, maturation and morphology of cell lines, and detection of infiltrative processes or malignancies. Findings are essential for diagnosis of blood disorders and marrow failure.
This document provides information about bone marrow aspiration and biopsy procedures. It discusses the history and development of the procedures, anatomy of bone and bone marrow, indications for bone marrow examinations, how to perform aspirations and biopsies, complications, interpretation of results including cellularity, hematopoiesis, iron stores, and reticulin, and examples of bone marrow findings in different conditions like infections, deficiencies, and cancers. Bone marrow examinations provide important diagnostic information for hematological diseases and disorders.
Bone marrow biopsy and aspiration provide qualitative and quantitative assessment of hematopoiesis. It can help make diagnoses of blood disorders like anemia and help stage diseases like lymphoma. The bone marrow has a structured organization with hematopoietic and stromal components. Biopsy and aspiration samples are analyzed microscopically after staining to evaluate cellularity, maturation of blood cell lineages, iron stores, and detect any abnormalities. This procedure helps diagnose conditions affecting the bone marrow including infections, storage diseases, and cancers.
Bone Marrow evaluation EVALUATION (PBS+BMA).pptxFereshtehAmeli1
Bone marrow examination is indicated for investigating unexplained blood abnormalities, diagnosing hematopoietic neoplasms, infectious disease workup, evaluating storage disorders, and staging lymphomas or other cancers. An adequate bone marrow aspirate requires at least 3 particles per slide and 4 slides total, while an adequate trephine biopsy is at least 1.6 cm long. Bone marrow specimens are evaluated for cellularity, morphology of cells, immunohistochemistry, cytogenetics, and iron staining as needed. Cell distributions and accessory structures like osteoblasts are also assessed.
The document discusses bone marrow examination, including indications for bone marrow aspiration and biopsy. It describes the procedures for bone marrow aspiration and biopsy, including appropriate sites depending on patient age. It also covers preparation of bone marrow smears, examination of cellularity and myeloid to erythroid ratio, and provides normal ranges for differential counts in adult bone marrow.
The document discusses bone marrow examination and preparation of bone marrow smears. It covers indications for bone marrow examination including diagnosis of hematological and non-hematological diseases. The common sites of bone marrow aspiration are described based on patient age. Procedures for bone marrow aspiration and trephine biopsy are outlined along with preparation of bone marrow films from aspirated samples.
This document provides information about bone marrow examination. It discusses:
1. Bone marrow is located within bone cavities and contains hematopoietic cells, sinusoids, fibroblasts, fat cells, and macrophages. There are two types - red (active) marrow and yellow marrow.
2. Hematopoietic cells are arranged between sinusoids and supported by fibroblasts. Erythroid precursors cluster near sinusoids while granulocytes are near bone. Megakaryocytes are near sinusoid walls.
3. Bone marrow aspiration and biopsy are the main examination methods and each provides different information. Aspiration assesses cell morphology while biopsy studies architecture and fibrosis. Indications
Dr shashi bansal approch to bone marrow examinationShashi Bansal
This document provides guidance on performing and interpreting bone marrow examinations. It discusses:
1. The importance of bone marrow examinations for diagnosing blood disorders when other tests are inconclusive.
2. The procedures involved in bone marrow examinations, including aspiration, biopsy, staining, and specialized testing.
3. How to analyze bone marrow samples under the microscope, including identifying cell types, assessing cellularity, iron content, fibrosis, and other features that provide diagnostic information.
Bone marrow aspiration & trephine biopsySanjeev Kumar
Bone marrow aspiration & trephine biopsy, Complication of BM Aspiration, Clinical significance, Indication of Bone Marrow Aspiration and Biopsy, Types Of Needles, Site for Bone Marrow Biopsy And Aspiration, types Of Smear for Bone Marrow, Advantages of Bone Marrow Trephine Biopsy
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 provides information about Pindborg tumor, also known as calcifying epithelial odontogenic tumor (CEOT). It defines CEOT as a locally invasive epithelial odontogenic neoplasm characterized by the presence of amyloid material that may become calcified. The document discusses the pathogenesis, histopathological features including epithelial cells, amyloid-like material and calcific deposits, immunohistochemical findings, differential diagnosis and treatment of CEOT. It also mentions the recurrence rate of CEOT is typically 10-15% but can be higher in certain variants.
Bone marrow examination is used to diagnose conditions like leukemia, multiple myeloma, and anemia. Bone marrow samples are obtained through aspiration or biopsy of the sternum, iliac crest, or tibia. Samples are prepared as bone marrow films which are stained and examined under a microscope. The cellularity, myeloid to erythroid ratio, and differential count of cell types in the bone marrow are assessed to evaluate for conditions like aplastic anemia or myeloproliferative disorders.
This document discusses animal models for studying vulnerable plaque and atherosclerosis. It describes how the chicken, pig, mouse, and rabbit models each have advantages and limitations for replicating human disease. The mouse has been genetically modified to better model plaque development. No current animal model fully captures plaque rupture seen in humans. The rabbit model with an embedded inflatable balloon shows promise for experimentally inducing plaque rupture.
This document discusses animal models for studying vulnerable plaque and atherosclerosis. It describes how the chicken, pig, mouse, and rabbit models each have advantages and limitations for replicating human disease. The mouse has been genetically modified to better model plaque development. No current animal model fully captures plaque rupture seen in humans. The rabbit model with an embedded inflatable balloon shows promise for experimentally inducing plaque rupture.
This document discusses animal models for studying vulnerable plaque and atherosclerosis. It describes how the chicken, pig, mouse, and rabbit models each have advantages and disadvantages for replicating different aspects of human disease. The mouse has been genetically modified to better model plaque development. No single animal model fully captures the human condition, but they provide insights into disease mechanisms. The document also notes that no current animal model replicates the plaque rupture seen in human heart attacks.
Parotid tumour n management dr karan r rawatKaran Rawat
Dr. Beth Eselm Finseyoum discusses parotid gland tumors and their treatment. The parotid gland is located in front of the ear and contains the facial nerve. Common tumors include pleomorphic adenoma and Warthin's tumor, which are usually benign. Mucoepidermoid carcinoma is the most common malignant tumor. Investigation may include ultrasound, CT scan and MRI. Treatment depends on tumor type, grade and extent, and may involve surgery such as parotidectomy or radiotherapy. Outcomes vary depending on tumor aggressiveness, with malignant tumors having a poorer prognosis.
Oral exfoliative cytology is a technique that examines cells collected from the oral mucosa. It can help detect undiagnosed diseases through a simple procedure and confirm suspected diseases without trauma. The document discusses the definition, role, indications, advantages and preparation methods of oral cytology. It describes how to make smears and various staining and fixation techniques used. Cytological classification systems and what normal and abnormal oral cells look like are presented. The uses of oral cytology in detecting oral cancer and newer diagnostic methods like cytomorphometry are also summarized.
Basal cell Adenoma and Canalicular Adenoma Doctor Faris Alabeedi MSc, MMedSc,...Doctor Faris Alabeedi
1. Basal cell adenoma and canalicular adenoma are benign salivary gland tumors composed of basaloid or ductal cells respectively.
2. Basal cell adenoma most commonly occurs in the parotid gland in elderly adults, while canalicular adenoma has a predilection for the minor salivary glands of the upper lip.
3. Histologically, basal cell adenoma demonstrates nests or cords of basaloid cells surrounded by a hyaline layer, whereas canalicular adenoma shows branching cords of ductal cells within a vascular stroma.
This document discusses bone marrow examination, including its purpose, types (aspiration and biopsy), indications, contraindications, procedure details, instruments used, complications, and findings in conditions like iron deficiency anemia and megaloblastic anemia. Bone marrow examination assesses hematopoiesis through aspiration or biopsy of the medullary cavity in bones. Aspiration is used to evaluate cellularity and differentials while biopsy allows evaluation of morphology, architecture, and focal lesions. Common sites include the iliac crest and sternum.
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.
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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
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2. BONE MARROW
Bone marrow examination is an indispensable
adjunct to the study of diseases of the blood
and may be the only way in which a correct
diagnosis can be made.
3. 1905 Pianese Trephine bx in an infant with Leishmaniasis.
1909 Pianese Tibia & femur marrow aspiration with
attached syringe.
1923 Seyfaith Surgical trephine to obtain marrow from
ribs & sternum.But there was excessive
bleeding.
1927 Airinkin Eliminated trephine complication by using
short lumbar needle.
1945 Vanderberg First obtained marrow from iliac creast.
1952 Bierman Used posterior Iliac creast for biopsy
HISTORY
4. 1. Bone marrow aspiration
2. Clot section
3. Bone marrow biopsy
4. Biopsy imprint smears
5. INDICATIONS OF BONE MARROW ASPIRATION
• Red cell disorders
• Leucocytic disorders
• Megakaryocyte and platelet disorders
• Myeloproliferative disorders and myelodysblastic
syndrome with BMB
• Paraproteinemias
• Infection
• Storage disorders
• Iron assessment
• Metastasis
8. 5. Unexplained leukoerythroblastic picture.
6. In suspected cases of multiple myeloma and serum
paraproteins.
7. Diagnosis and staging of
Non hodgkin’s lymphoma
Hodgkin’s lymphoma
Malignancy
Metastatic carcinoma
Small round cell tumors of childhood
8. Stromal changes
Fibrosis
Necrosis
Gelatinous marrow transformation
9. 9. Pyrexia of unknown origin
10. Focal lesions –Metastasis, Granuloma
11.Amyloidosis
12.Metabolic bone diseases
13.To assess the mineralisation front and appositional
growth after tetracycline labeling
10. CONTRADICATIONS
Sternal aspirate-osteoporosis and children
Biopsy in coagulopathies
(For aspiration factor replacement therapy
prior to procedure and observation should be
done for next 24-48 hrs.)
11. COMPLICATIONS
Hemorrhage
(Risk factors- coagulopathies,
myeloproliferative disorders, aspirin and
warfarin therapy, thrombocytopenia, DIC, liver
disease and disease)
Pain
Infection
Perforation of major vessels
Risk of general anaesthesia and sedation.
12. BONE MARROW ASPIRATE V/S BIOPSY
Aspiration biopsy and trephine biopsy are
complementary to each other.
Better cytological detail.
More range for
cytochemical stains,
flowcytometry and IHC.
Ideal for cytogenetics
and molecular genetics.
Topographical details,
cellularity and
infiltration.
Less range.
Can be used for both.
13. Dry tap in fibrosis
Can be performed alone in
iron deficiency anemia,
anemia of chronic
disease , megaloblastic
anemia and acute
leukaemia.
Less painful
Essential for diagnosis
in dry tap.
Helpful for aplastic
hypoplastic anaemia,
lymphoma, metastatic
carcinoma,
myeloproliferative
neoplasms and
diseases of the bones.
More painful
14. IMPORTANCE OF TOUCH/ IMPRINT
SMEAR
Gives cytological details when aspirate is not
obtained.
Shows more neoplastic cells than aspirate.
Can show marrow infiltration , not seen in
aspirate
15. IMPORTANCE OF CLOT SECTION
Assessment of bone marrow cellularity.
For detecting granuloma and tumour infiltrates
complementary to biopsy.
No decalcification associated nucleic acid or
protein damage.
16. SITE FOR ASPIRATION
1. Posterior superior iliac spine- most preferred.
2. Anterior superior iliac spine. (The iliac spines have the
advantage that if no material is aspirated, a trephine biopsy can be
performed immediately.)
3. Sternum (abt 1cm above the sternomanubrial
angle,to one side of midline). (avoided in babies)
4. Medial aspect of tibia just below tibial tubercle
(small babies).
5. Spinous process of vertebrae
17. 1. Posterior superior iliac spine- most
preferred.
2. Anterior superior iliac spine.
The posterior iliac spine is said to provide samples that are
longer and larger, and the aspiration is less uncomfortable for
the patient.
18. Needles should be stout and made of hard
stainless steel, about 7-8 cm in length, with
a well-fitting stilette, and they must be
provided with an adjustable guard.
1. Jamshidi needle
2. Islam needle
3. Salah needle
4. Klima needle
19. If larger specimens are needed, trephine
needles that have bores of 4-5 mm may be
used.
Other needles occasionally used for trephine
biopsy specimens are a 2-mm bore
“microtrephine” needle and a Vim–Silverman
needle.
However, compared with other needles, these
yield smaller specimens of marrow that are
prone to fracturing.
20. a: STYLET b : BORE c : PROBE
The tip gradually widens for next 2cm and then uniform bore the needle.The
marrow biopsy does not get compressed because of narrow cutting edges and
hence cell morhology is well made out.
JAMSHIDI NEEDLE
21. ISLAM
NEEDLE
A modified version of the Islam needle has multiple holes in the distal
portion of the shaft in addition to the opening at the tip to overcome
sampling error when the marrow is not uniformly involved in a
pathological lesion.
23. PROCEDURE
Consent- an written consent should be taken from
patient.
• An appropriate clinical history should accompany
the bone marrow, as they relate to possible
findings within the bone marrow examination.
24. It is useful to know relevant laboratory data
such as Iron studies, Folate or Vitamin B-12
studies, transfusion therapy, hematinics or
history of chemotherapy.
The physician’s clinical impression should be
included on the form.
Lignocaine sensitivity test should be done.
25. Either aspirate or biopsy may be performed first.
But the two should be performed through the
same incision, approx. 0.5-1 cm away from the
other.
This is done to avoid clotting of aspirate and
hemorrhagic or damaged biopsy.
It is recommended that the aspirate and biopsy
be obtained using respective needles separately,
and not through a trephine needle.
26. • Operator should wear Surgical gloves
• Skin at the site should be cleaned with 70%
alcohol or 0.5% chlorhexidine.
• Infiltrate the skin, s/c tissue and periosteum
over the site with 2-5ml of lignocaine.
• Children are either sedated or given general
anaesthesia.
27. With boring movement pass the needle
perpendicularly at center of PSIS.
When bone has been penetrated, remove the
stilette, attach 30 ml syringe and suck up
marrow contents (0.3ml) for making smears
immediately.
28. If large sample is needed for cytogenetics and
immunophenotyping attach another 5 or 10 ml
syringe and aspirate.
(should be kept in preservative free heparin than in EDTA)
• Fix the slides in absolute methanol (20 min) as
soon as they are thoroughly dry for subsequent
staining by a Romanowsky method or Perls' stain
for iron.
29. The preparation can be considered satisfactory
only when marrow particles and free marrow
cells can be seen in stained films.
These films are also suitable for cytochemical
staining.
30. Some material (clots) can be preserved in
fixative rather than anticoagulant for preparation
of histological sections.
(clot section).
Clot is processed same as bone marrow biopsy
but without decalcification
• Peripheral blood sample is also taken along with
it.
31. • Dry tap- Failure to aspirate marrow (suggests
bone marrow fibrosis or infiltration.)
• If there has been a “dry tap,” insert the
stilette into the needle and push any material
in the lumen of the needle onto a slide; in
lymphomas and carcinomas, especially,
sufficient material can be obtained to make a
diagnosis.
• CT–guided marrow sampling-obese, in whom
it is difficult to locate the iliac spine.
33. EXAMINATION OF ASPIRATED BONE
MARROW
Low power (10x)
Determine cellularity
Identify megakaryocytes and note
morphology and maturation sequence
(higher power may be needed for smaller immature megakaryocytes and
micromegakaryocytes).
Look for clumps of abnormal cells
(higher power needed to examine content and morphology of clumps)
Identify macrophages
(higher power for evidence of haemophagocytosis, malaria pigment, and
bacterial or fungal infections that may be present in the cytoplasm)
34. High power (40x,100x oil immersion)
Identify all stages of maturation of myeloid
and erythroid cells
Maturation abnormalities are noted
Determine the myeloid:erythroid ratio
Perform a differential count using the
categories erythroid, myeloid, lymphoid, plasma
cell, and “others,” simultaneously noting any
morphological abnormalities.
Look for areas of bone marrow necrosis
Assess the iron content
48. OSTEOBLAST have basophillic cytoplasm,extruding nucleus and regular
chromatin with 1-4 nucleoli. Can be distinguished from plasma cells by their larger
size and the position of the Golgi zone, which is not immediately adjacent to the
nucleus.
49. An osteoclast; note the highly granular cytoplasm and the multiple nuclei (2-
100)which are uniform in size and have indistinct, medium - sized, single nucleoli.
MGG × 100.
50. Mast cells on bone marrow.
With PAS stain they display magenta coloured granules
51. Mature megakaryocyte having loblated nucleus and pink granular
cytoplasm..platelets are formed by budding of the cytoplasm which are
shed in the circulation.
52. Aspirate of normal BM: a macrophage containing granular and refractile debris
and several normoblast nuclei. MGG × 100.
53. Aspirate of non - infiltrated BM from a patient with Hodgkin lymphoma: a mature
megakaryocyte exhibiting emperipolesis. MGG× 100.
54. Grading of bone marrow storage iron
0 No stainable iron
1+ Small iron particles just visible in reticulum
cells using an oil objective
2+ Small, sparse iron particles in reticulum cells,
visible at lower power
3+ Numerous small particles in reticulum cells
4+ Larger particles with a tendency to
aggregate into clumps
5+ Dense, large clumps
6+ Very large clumps and extracellular iron
55. Grading for iron on bone marrow aspirate
Grade 1 Grade 2 GRADE 3
1+ Small iron particles just visible in reticulum cells using an oil objective
2+ Small, sparse iron particles in reticulum cells, visible at lower power
3+ Numerous small particles in reticulum cells
56. Iron grading
4+ Larger particles with a tendency to aggregate into clumps
5+ Dense, large clumps
6+ Very large clumps and extracellular iron
GRADE 4 GRADE 5 GRADE 6
61. BIOPSY (Procedure)
• The trephine specimen is obtained by inserting
the biopsy needle into the bone and using a to-
and-fro rotation to obtain a core of tissue.
• If an aspirate has been performed first the
needle should be inserted through the same
incision but the needle should be advanced at a
slightly different angle.
62. • The bony core is gently dabbed or rolled
across the slide to form imprint smears, which
is then fixed and stained as for bone marrow
aspiration smears.
This allows immediate examination of cells that fall out of the
specimen onto the slide and may provide a diagnosis several
days before the trephine biopsy specimen has been
processedaspirate. and also useful in dry
• Bilateral trephine biopsies may be performed
to increase the yield of detecting focal lesions.
63. Length at least 1.5 cm
At least 10 partialy preserved trabecular
spaces seen
Sections of 3-4 micron in thickness cut at a
distance of 50 micron each.
(WHO Classification of tumours of
haematopoietic and lymphoid tissue 2008)
64. FIXATION:
Biopsy is fixed in 10% neutral buffered
formalin for 6 hrs.(ICSH preferred)
Other fixatives: Zinc formaldehyde
Acetic acid formalin
Isotonic buffered formalin
B5: not used due to presence of mercuric
chloride (poisonous)
Bouin’s fixative: not used in some countries,
due to presence of picric acid.
65. It is important to ensure that the formol -
saline is not left for long periods at ambient or
high temperature before being used because
formic acid and formalin pigment may be
produced.
The reactivity of antibodies used for
immunohistochemical staining may also be
affected by the choice of fixative (use of
Bouin’s solution is particularly limiting) and
Zenker’s fixative can destroy chloroacetate
esterase activity.
66. B5 fixative- if fixation lasts for more than 6
hours hardening of the tissue can make it
difficult to cut sections.
67. Decalcifier agent volume to tissue volume should
be 20:1
1. 10-15% EDTA( 1-3 days) preserves morphology,
enzymes and immunological epitopes. ICSH
recommended
2. 10% Formic acid( 2-3 days) causes more tissue
distortion.
68. 3. 5-10% nitric acid(3-6 hrs) used for urgent
processing. (nitric acid can cause
megakaryocytes to give positive reactions
with antibodies to CD34 and affect IHC.)
4. In our lab-EDTA.
69. DECALCIFICATION :PITFALLS
It chelates storage iron.
Affects the morphology and cytological details.
Affects ability to perform immunohistochemistry
and to retrieve material for molecular analysis.
70. Factors affecting decalcification
Conc. of decalcifying agent.
Temperature
Agitation
Age of patient
Type of bone
Size of spicules
71. Decalcification endpoint
Specimen radiography (FAXITRON,most
reliable)
Chemical method
Calcium oxalate test
(5ml of used decalcifying agent+Ca(OH)+ 5
mlsat. ammonium oxalate)
Physical method
Bubble test
72. :
Causes shrinkage and loss of some cellular
detail but better for IHC.
:
Glycol methacrylate or methyl methacrylate
used
Finer sections with better cellular details.
No decalcification required so used for
metabolic bone diseases.
73. SECTIONING
At least 6 sections should be cut at 3 levels
into cross sectional diameter of core-
25%
50%
75%
Additional sections needed for IHC or
histochemical stains
75. EVALUATION OF BONE MARROW BIOPSY
4x or 10x
Adequacy
Cellularity
Pattern
Presence of focal lesions
Megakaryocyte number
Abnormal cell clusters and location
Bone structure
Osteoclastic and osteoblastic activity.
76. 40x
Assess haemopoietic cells(erythroid, myeloid,
megakaryocytes, lymphoid cells, plasma cells
and macrophages) and cytological details.
Oil immersion
For finer cytological details (eg.intracellular
granules, organisms.)
77. Normal topography
Myeloid cells
Paratrabecular
Mature cells towards centre
Erythroid cells
Centre in colonies
Megakaryocytes
Centre around sinusoids
78. Monocyte precursors:
Not recognizable.
Some can be seen randomly distributed.
Lymphoid precursors:
Seen in periarteriolar region.
Stroma
Fat cells, fibroblasts, reticulin fibres
79. Osteocytes:
Seen in bony
lacunae.
Osteoblasts:
Seen lining the
trabeculae.
Osteoclasts:
Seen in howship’s
lacunae
.
80. Topography of cells
Bone marrow is highly organised structure with haemopoietic
elements maturing in different micro-anatomical sites.
89. Mast cells in immunoperoxidase stain
Usually seen in giemsa . Present irregularly in medullary cavity
90. Quantification of bone marrow reticulin
and collagen
0 No reticulin fibres demonstrable
1 Occasional fine individual fibres and foci of
fine fibre network
2 Fine fibre network throughout most of the
section; no coarse fibres
3 Diffuse fibre network with scattered thick
coarse fibres but no mature collagen
4 Diffuse often coarse fibre network with areas
of collagenization
91. grade 0 grade 1
Fine fibre network throughout
most of the section; no
Occasional fine individual
fibres and foci of fine fibre
network
grade 2,
92. grade 3,
grade 4,
Diffuse fibre network with scattered
thick coarse fibres but no mature
collagen
Diffuse often coarse fibre network with
areas
94. SUPPLEMENTARY INVESTIGATIONS
- Immunohistochemistry- for demonstration of
antigens in biopsy
Ex.- CD 34, CD 45, Lysozyme, MPO, CD 68,
CEA etc.
- Cytogenetic analysis- for chromosomal
rearrangements
- Molecular genetics- by PCR, RTPCR
FISH
95. Bone marrow biopsy report (ICSH guidelines
2008):
Name of institution
Unique specimen identifier
Details of patient- name,age,gender, contact details
Name of responsible physician
Name of requesting doctor
Date of procedure
Clinical history, examination,therapy
Indication of bone marrow examination
Procedure performed
Site of procedure
96. Signature and date of report
Length of biopsy core
Adequacy and appearance of the core
Percentage and pattern of cellularity
Bone architechture
Location, number,morphology and pattern of differentiation
of
1. Erythroid
2. Myeloid
3. Megakaryocytic
4. Lymphoid
5. Plasma cells
6. Macrophages
Abnormal cells
Reticulin stain
IHC
Other Ix
97. Turn around time
Varies from lab to lab
24 hrs in case of microwave decalcification and as
long as 1 week if EDTA based decalcifying agent
is used.
5 days for less urgent cases
Additional 1-2 days if IHC is required
In our lab aspiration is reported within 24hrs and
biopsy within 4-5 days.
99. Quality assurance
North America and Europe have well trenched
external quality assessment for histopathology
which includes bone marrow biopsy.
Situation in India is less advanced due to
vast unorganised structure of private labs
and
highly variable training, technological facilities
and reporting styles.
100. TAKE HOME MESSAGE
Biopsy of the bone marrow is an
indispensable adjunct to the study of diseases
of the blood and may be the only way in
which a correct diagnosis can be made.
A minimum definition of an adequate bone
marrow biopsy specimen stipulates a length of
atleast 1.5 cm, 5-6 marrow spaces and no
aspiration, crush or processing artifacts.
101. • Not to assess histology in isolation, ideally reported
along with bone marrow aspirate and peripheral
smear and preferrably reported by same pathologist.
• There should be an integrated approach in reporting
which includes
Clinical history
Laboratory investigations,
Imaging information
IHC wherever applicable.