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Pathology Practical Book
Harsh Mohan
SECTION ONE: TECHNIQUES IN PATHOLOGY
EXERCISE 1:
Microscopy of Various Types
LIGHT MICROSCOPE
Stand
Body
Body Tubes
Stage
Knobs for Coarse and Fine Adjustment
Optical System
Eye Piece
Objectives
Condenser
Light/Illumination System
Magnification and Resolving Power of Light Microscope
How to Use a Light Microscope
OTHER TYPES OF MICROSCOPY
Dark Ground Illumination (DGI)
Principle
Polarising Microscope
Principle
Fluorescent Microscope
Principle
Electron Microscope
Principle
Transmission Electron Microscopy (TEM)
Scanning Electron Microscopy (SEM)
RECENT ADVANCES IN MICROSCOPY
Image Analysers and Morphometry
Telepathology (Virtual Microscopy)
EXERCISE 2:
Histopathology Techniques and Routine Staining
FIXATION
Types of Fixatives
Formalin
Glutaraldehyde
Bouin's Fluid (Picric acid)
Carnoy's Fixative (Alcohol)
Osmium tetraoxide
DEHYDRATION
CLEARING
IMPREGNATION
TISSUE PROCESSORS
EMBEDDING AND BLOCKING
SECTION CUTTING (MICROTOMY)
Procedure for Microtomy
ROUTINE STAINING (H & E)
Haematoxylin
Procedure for Staining
EXERCISE 3:
Frozen Section and Special Stains
FROZEN SECTION
Merits
Demerits
Methods for Frozen Sections
Freezing Microtome using CO2 Gas
Refrigerated Microtome (Cryostat)
Staining of Frozen Sections
Rapid H & E Staining
Toluidine Blue Staining
SPECIAL STAINS
Sudan Black/Oil Red O
Procedure for Oil Red O Staining
Result
van Gieson
Result
Masson's Trichrome
Result
Reticulin
Result
Congo Red
Result
Periodic Acid-Schiff (PAS)
Result
Methyl Violet
Result
Prussian Blue/Perl's Reaction
Result
SECTION TWO: CLINICAL PATHOLOGY
EXERCISE 4:
Urine Examination I: Physical and Chemical
A. ADEQUACY OF SPECIMEN
Specimen Collection
Methods of Preservation of Urine
B. PHYSICAL EXAMINATION
Volume
Colour
Odour
Reaction/pH
Specific Gravity
1. Urinometer
Procedure
2. Refractometer
3. Reagent Strip Method
Significance of Specific Gravity
C. CHEMICAL EXAMINATION
Tests for Proteinuria
Qualitative Tests for Proteinuria
1. Heat and Acetic Acid Test
2. Sulfosalicylic Acid Test
3. Heller's Test
4. Reagent Strip Method
Quantitative Estimation of Proteins in Urine
1. Esbach's albuminometer method
2. Turbidimetric method
Test for Bence-Jones Proteinuria
Test for Glucosuria
Qualitative Tests
1. Benedict's Test
Procedure
Interpretation
2. Reagent Strip Test
Quantitative Test for Glucose
Causes of Glucosuria
Tests for Ketonuria
Tests for Ketonuria
1. Rothera's Test
Procedure
2. Gerhardt's Test
Procedure
3. Reagent Strip Test
Causes of ketonuria
Test for Bile Derivatives in Urine
Tests for Bile Salts
1. Hay's Test
Procedure
2. Strip Method
Causes for bile salts in urine:
Tests for Urobilinogen
1. Ehrlich's Test
Procedure
2. Reagent Strip Test
Tests for Bilirubin (Bile Pigment) in Urine
1. Fouchet's Test
Procedure
2. Foam Test
Procedure
3. Reagent Strip Test
Causes of bilirubinuria
Tests for Blood in Urine
1. Benzidine Test
Procedure
2. Orthotoluidine Test
Procedure
3. Reagent Strip Test
Causes of blood in urine
AUTOMATED URINALYSIS
EXERCISE 5:
Urine Examination II: Microscopy
COLLECTION OF SAMPLE
PREPARATION OF SEDIMENT
EXAMINATION OF SEDIMENT
1. Examination of Cells
RBCs
Physiological
Pathological
WBCs
Pathological
Epithelial Cells
2. Casts
Hyaline Cast
Red Cell Cast
Leucocyte Cast
Granular Casts
Waxy Casts
Fatty Cast
Epithelial Cast
Pigment Cast
Crystals
Crystals in Acidic Urine
i) Calcium Oxalate
ii) Uric Acid
iii) Amorphous Urate
iv) Tyrosine
v) Cystine
vi) Cholesterol Crystals
vii) Sulphonamide
Crystals in Alkaline Urine
i) Amorphous Phosphate
ii) Triple Phosphate
iii) Calcium Carbonate
iv) Ammonium Biurate
4. Miscellaneous Structures in Urine
i) Spermatozoa
ii) Parasites
iii) Fungus
iv) Tumour Cells
AUTOMATION IN URINE ANALYSIS
Urine Strip Analysers
Flow Cytometry
EXERCISE 6:
Semen Analysis
A. SAMPLE COLLECTION
B. GROSS EXAMINATION
C. MICROSCOPIC EXAMINATION
Motility
Count
Morphology
D. CHEMICAL EXAMINATION
Fructose Test
Procedure
Acid Phosphatase Test
E. IMMUNOLOGICAL ASSAYS
F. MICROBIOLOGICAL ASSAYS
G. SPERM FUNCTION TESTS
EXERCISE 7:
Examination of CSF
NORMAL COMPOSITION OF CSF
A. SPECIMEN COLLECTION
B. MICROSCOPIC EXAMINATION
Total Leucocyte Count
Manual Method
Automated Method
Differential Leucocyte Count
Conditions Causing Increased Neutrophils in CSF
Conditions Causing Increased Lymphocytes in CSF
Conditions Causing Plasma Cells in CSF
Conditions Causing Lymphocytes and Monocytes in CSF
Conditions Causing Malignant Cells in CSF
C. CHEMICAL EXAMINATION
D. MICROBIOLOGICAL EXAMINATION
E. IMMUNOLOGICAL EXAMINATION
SECTION THREE: GENERAL PATHOLOGY
EXERCISE 8:
Degenerations
VACUOLAR NEPHROPATHY
HYALINE CHANGE IN LEIOMYOMA
MYXOID DEGENERATION IN GANGLION
EXERCISE 9:
Intracellular Accumulations
FATTY CHANGE LIVER
MELANIN PIGMENT IN NAEVUS
ANTHRACOTIC PIGMENT IN LUNG
BROWN ATROPHY HEART
EXERCISE 10:
Amyloidosis
AMYLOIDOSIS KIDNEY
AMYLOIDOSIS SPLEEN
AMYLOIDOSIS LIVER
EXERCISE 11:
Necrosis
COAGULATIVE NECROSIS (INFARCT) KIDNEY
LIQUEFACTIVE NECROSIS (INFARCT) BRAIN
CASEOUS NECROSIS LYMPH NODE
ENZYMATIC FAT NECROSIS PANCREAS
EXERCISE 12:
Gangrene and Pathologic Calcification
WET GANGRENE BOWEL
DRY GANGRENE FOOT
MONCKEBERG'S ARTERIOSCLEROSIS
EXERCISE 13:
Derangements of Body Fluids
PULMONARY OEDEMA
CVC LUNG
CVC LIVER
CVC SPLEEN
EXERCISE 14:
Obstructive Circulatory Disturbances
THROMBUS ARTERY
PALE INFARCT SPLEEN
HAEMORRHAGIC INFARCT LUNG
EXERCISE 15:
Inflammation
ABSCESS LUNG
CHRONIC INFLAMMATORY GRANULATION TISSUE
TUBERCULOUS LYMPHADENITIS
EXERCISE 16:
Tuberculous Granulomatous Inflammation
FIBROCASEOUS TUBERCULOSIS LUNG
TUBERCULOSIS INTESTINE
MILIARY TUBERCULOSIS LUNG
MILIARY TUBERCULOSIS SPLEEN
EXERCISE 17:
Other Granulomatous Inflammations
LEPROMATOUS LEPROSY
TUBERCULOID LEPROSY
SARCOIDOSIS LUNG
EXERCISE 18:
Specific Infections and Infestations I
ACTINOMYCOSIS SKIN
MADURA FOOT
ASPERGILLOSIS LUNG
EXERCISE 19:
Specific Infections and Infestations II
RHINOSPORIDIOSIS NOSE
CYSTICERCOSIS SOFT TISSUE
HYDATID CYST LIVER
EXERCISE 20:
Growth Disorders
TESTICULAR ATROPHY
CARDIAC HYPERTROPHY
REACTIVE HYPERPLASIA LYMPH NODE
SQUAMOUS METAPLASIA CERVIX
EXERCISE 21:
Neoplasia I
SQUAMOUS CELL PAPILLOMA
SQUAMOUS CELL CARCINOMA
MALIGNANT MELANOMA
BASAL CELL CARCINOMA
EXERCISE 22:
Neoplasia II
LIPOMA
PLEOMORPHIC RHABDOMYOSARCOMA
METASTATIC CARCINOMA LYMPH NODE
METASTATIC SARCOMA LUNG
SECTION FOUR: SYSTEMIC PATHOLOGY
EXERCISE 23:
Blood Vessels and Lymphatics
ATHEROMA CORONARY ARTERY
CAPILLARY HAEMANGIOMA SKIN
CAVERNOUS HAEMANGIOMA LIVER
LYMPHANGIOMA
EXERCISE 24:
Heart
BACTERIAL ENDOCARDITIS
HEALED MYOCARDIAL INFARCT
CHRONIC ISCHAEMIC HEART DISEASE
FIBRINOUS PERICARDITIS
EXERCISE 25:
Respiratory System I
LOBAR PNEUMONIA—ACUTE CONGESTION STAGE
LOBAR PNEUMONIA—RED HEPATISATION STAGE
LOBAR PNEUMONIA—GREY HEPATISATION STAGE
BRONCHOPNEUMONIA
EXERCISE 26:
Respiratory System II
EMPHYSEMA
BRONCHIECTASIS
SMALL CELL CARCINOMA LUNG
SQUAMOUS CELL CARCINOMA LUNG
EXERCISE 27:
GIT I
AMELOBLASTOMA
PLEOMORPHIC ADENOMA
PEPTIC ULCER
ULCERATIVE COLITIS
EXERCISE 28:
GIT II
ACUTE APPENDICITIS
JUVENILE POLYP RECTUM
ADENOCARCINOMA STOMACH
MUCINOUS ADENOCARCINOMA COLON
EXERCISE 29:
Liver and Biliary System I
ACUTE VIRAL HEPATITIS
ALCOHOLIC HEPATITIS
SUBMASSIVE NECROSIS OF LIVER
EXERCISE 30:
Liver and Biliary System II
CIRRHOSIS LIVER
HEPATOCELLULAR CARCINOMA
CHRONIC CHOLECYSTITIS WITH CHOLELITHIASIS
CARCINOMA GALLBLADDER
EXERCISE 31:
Urinary System I
ACUTE GLOMERULONEPHRITIS (GN)
RAPIDLY PROGRESSIVE GLOMERULONEPHRITIS (RPGN)
CHRONIC GLOMERULONEPHRITIS
CHRONIC PYELONEPHRITIS
EXERCISE 32:
Urinary System II
DIABETIC NEPHROSCLEROSIS
RENAL CELL CARCINOMA
WILMS’ TUMOUR
TRANSITIONAL CELL CARCINOMA
EXERCISE 33:
Lymphoid System
NON-HODGKIN'S LYMPHOMA (NHL)
HODGKIN'S DISEASE (HD)
HD-Nodular Sclerosis Type
HD-Mixed Cellularity Type
EXERCISE 34:
Male Reproductive System and Prostate
SEMINOMA TESTIS
NODULAR HYPERPLASIA PROSTATE
ADENOCARCINOMA PROSTATE
EXERCISE 35:
Female Reproductive System I
SIMPLE (CYSTOGLANDULAR) HYPERPLASIA
HYDATIDIFORM MOLE
INVASIVE CERVICAL CANCER
EXERCISE 36:
Female Reproductive System II
SEROUS OVARIAN TUMOURS
Serous Cystadenoma Ovary
Papillary Serous Cystadenocarcinoma Ovary
MUCINOUS OVARIAN TUMOURS
Mucinous Cystadenoma Ovary
BENIGN CYSTIC TERATOMA OVARY
EXERCISE 37:
Breast
FIBROADENOMA
SIMPLE FIBROCYSTIC CHANGE
INFILTRATING DUCT CARCINOMA-NOS
EXERCISE 38:
Thyroid
FOLLICULAR ADENOMA
NODULAR GOITRE
HASHIMOTO'S THYROIDITIS
PAPILLARY CARCINOMA
EXERCISE 39:
Bones and Joints I
CHRONIC OSTEOMYELITIS
TUBERCULOUS OSTEOMYELITIS
OSTEOCHONDROMA
OSTEOCLASTOMA
EXERCISE 40:
Bones and Joints II
EWING'S SARCOMA
OSTEOSARCOMA
CHONDROSARCOMA
EXERCISE 41:
Nervous System
ACUTE PYOGENIC MENINGITIS
MENINGIOMA
SCHWANNOMA
ASTROCYTOMA
SECTION FIVE: CYTOPATHOLOGY
EXERCISE 42:
Basic Cytopathologic Techniques
EXFOLIATIVE CYTOLOGY
Applications of Exfoliative Cytology
Female Genital Tract
Respiratory Tract
Gastrointestinal Tract
Urinary Tract
Body Fluids
Buccal Smears for Sex Chromatin
Fixation of Smears in Exfoliative Cytology
Staining of Smears in Exfoliative Cytology
Papanicolaou Stain
H & E Stain
Romanowsky Stain
ASPIRATION CYTOLOGY
Applications of FNA
I. Palpable Mass Lesions in:
II. Non-Palpable Mass Lesions in:
Procedure for FNA
Method
Radiological Imaging Aids for FNA
Advantages of FNA over Surgical Biopsy
IMPRINT CYTOLOGY
EXERCISE 43:
Exfoliative Cytology
PAP SMEAR—INFLAMMATORY
PAP SMEAR—CARCINOMA CERVIX
FLUID CYTOLOGY FOR MALIGNANT CELLS
EXERCISE 44:
Fine Needle Aspiration Cytology
FNA FROM TUBERCULOUS LYMPHADENITIS
FNA FROM FIBROADENOMA BREAST
FNA FROM DUCT CARCINOMA BREAST
SECTION SIX: HAEMATOLOGY
EXERCISE 45:
Haemoglobin Estimation—Various Methods
METHODS FOR ESTIMATION OF HAEMOGLOBIN
Cyanmet Hb Method
Procedure
Calculations
Advantages
Disadvantages
Oxyhaemoglobin Method
Procedure
Advantages
Disadvantages
Electronic Counter Method
Disadvantage
Direct Reading Electronic Haemoglobinometers
Disadvantage
Sahli's Method
Procedure
Advantages
Disadvantages
Normal Values of Hb
Errors in Haemoglobinometry
QUALITY CONTROL IN HAEMOGLOBIN ESTIMATION
EXERCISE 46:
Counting of Blood Cells
WBC COUNT
Methods
Visual Haemacytometer Method
Procedure
Calculations
Precautions
Electronic Method
Advantages
Disadvantages
Normal Range for WBC count
Causes of Abnormal Leucocyte Count
RBC COUNT
Methods for RBC Counts
Visual Haemacytometer Method
Diluting fluids
Compostion of Hayem's fluid
Composition of Dacie's fluid
Procedure
Calculations
Electronic Method
Advantages
Disadvantages
Normal Range for RBC Count
Cause of Decreased RBC Count
Cause of Increased RBC Count
PLATELET COUNT
Methods for Counting Platelets
Visual Method
Procedure
Calculations
Rough Visual Method for Platelet Counting
Electronic Method
Disadvantages
Normal Platelet Count
Conditions causing abnormal platelet counts
Conditions causing Thrombocytopenia
Conditions Causing Thrombocytosis
EXERCISE 47:
Reticulocyte Count
RETICULOCYTES
METHODS FOR COUNTING OF RETICULOCYTES
Visual Method
Procedure
Counting of Reticulocytes
Calculations
Automated Method
Normal Values
Abnormal Counts
Increased count (Reticulocytosis)
Decreased counts (Reticulocytopenia)
EXERCISE 48:
Preparation of Peripheral Blood Film, Staining and DLC
THIN BLOOD FILM
Slide Method
Procedure
Qualities of a Good Blood Film
Parts of a Thin Blood Film
Cover Glass Method
Procedure
Spin Method
Procedure
THICK BLOOD FILM
VARIOUS STAINS FOR PBF
Staining of Thin Blood Film
Leishman Stain
Procedure for staining
Giemsa Stain
Procedure
Staining of Thick Smear
Autostainers
Precautions in Staining of PBF
EXAMINATION OF PBF FOR DLC
MORPHOLOGIC IDENTIFICATION OF MATURE LEUCOCYTES
Polymorph (Neutrophil)
Lymphocyte
Monocyte
Eosinophil
Basophil
EXERCISE 49:
DLC in Cases with Leucocytosis
VISUAL COUNTING
AUTOMATED COUNTING
PATHOLOGIC VARIATIONS IN DLC
Neutrophils
Lymphocytes
Monocytes
Eosinophils
Basophil
EXERCISE 50:
ESR, PCV (Haematocrit) and Absolute Values
ERYTHROCYTE SEDIMENTATION RATE (ESR)
Principle
Mechanism of ESR
Phases in ESR
Methods of ESR
1. Westergren's Method
Procedure
Normal values
Advantages
Disadvantages
2. Wintrobe's Method
Procedure
Normal values
Advantages
Disadvantages
3. Micro ESR Method
Procedure
4. Automated ESR Method
Clinical Significance of ESR
Diagnostic Uses
Monitoring Prognosis of Diseases
PACKED CELL VOLUME (PCV) OR HAEMATOCRIT
Methods for Estimation of PCV
1. Macro (Wintrobe's) Method
Procedure
Advantages of Macro Method
2. Microhaematocrit Method
Procedure
Advantages of Micro Method
Sources of Errors in Macro and Micro Methods
3. Electronic Method
Clinical Significance of PCV
ABSOLUTE VALUES
1. Mean corpuscular volume (MCV)
2. Mean corpuscular haemoglobin (MCH)
3. Mean corpuscular haemoglobin concentration (MCHC)
Significance
EXERCISE 51:
Screening Tests for Bleeding Disorders
BLEEDING TIME
Principle
Methods for Bleeding Time
1. Finger Tip Method
Procedure
Disadvantages
2. Duke's Method
Procedure
Advantages of the method
3. Ivy's Method
Procedure
Advantages of the method
Clinical Application of Bleeding Time
CLOTTING TIME
Whole Blood Clotting Time
Methods
1. Capillary Tube Method
Procedure
Disadvantages
Advantages
2. Lee and White Method
Procedure
Advantages
Disadvantages
Sources of Error
Clinical Applications
EXERCISE 52:
Blood Grouping
ABO SYSTEM
Principle
Methods for ABO Grouping
1. Slide Method
2. Tube Method
Cell Grouping (Fig. 52.2,A)
Serum Grouping (Fig. 52.2,B)
Advantages of Tube Method Over Slide Method
Note
RHESUS (Rh) SYSTEM
Methods for Rh Grouping
1. Slide Method
2. Tube Method
Note
Incidence of ABO Blood Groups in India
Importance of Blood Grouping
EXERCISE 53:
Peripheral Blood Film Examination in Anaemias
PLAN OF INVESTIGATION FOR ANAEMIA
Haemoglobin Estimation
Peripheral Blood Film (PBF) Examination
Variation in Size (Anisocytosis)
Variation in Shape (Poikilocytosis)
Inadequate Haemoglobin Formation (Hypochromasia)
Compensatory Erythropoiesis
Miscellaneous Changes
Red Cell Indices
Leucocyte and Platelet Count
Reticulocyte Count
Bone Marrow Examination
Biochemical Tests
PERIPHERAL BLOOD FILM IN IRON DEFICIENCY ANAEMIA (IRON DEFICIENCY ANAEMIA)
PERIPHERAL BLOOD FILM IN MACROCYTIC ANAEMIA (MEGALOBLASTIC ANAEMIA)
PERIPHERAL BLOOD FILM IN HAEMOLYTIC ANAEMIA (THALASSAEMIA)
EXERCISE 54:
Blood Smear Examination in Leukaemias
PBF IN ACUTE LEUKAEMIAS: AML
PBF IN CHRONIC MYELOID LEUKAEMIA
PBF IN CHRONIC LYMPHOCYTIC LEUKAEMIA
EXERCISE 55:
Haemoparasites in Blood
PBF IN MALARIAL PARASITE
PBF IN FILARIASIS
BONE MARROW IN LEISHMANIASIS
EXERCISE 56:
Bone Marrow Examination
BONE MARROW ASPIRATION
TREPHINE BIOPSY
SECTION SEVEN: AUTOPSY PATHOLOGY
EXERCISE 57:
Introduction to Autopsy Protocol
INTRODUCTION
AUTOPSY PROTOCOL
A. Prosection
B. Gross Examination of Organs
C. Autopsy Report
EXERCISE 58:
Clinicopathological Conference (CPC) and About CD on CPCs
CLINOCOPATHOLOGICAL CONFERENCE (CPC)
ABOUT CD ON CPCS
CPC 1: Case of secondary systemic amyloidosis
CPC 2: Case of septic shock with DIC
CPC 3: Case of miliary tuberculosis with TBM
CPC 4: Case of RHD with infective endocarditis
CPC 5: Case of cirrhosis with portal hypertension
CPC 6: Case of brochogenic carcinoma
CPC 7: Case of long-standing inflammatory bowel disease
CPC 8: Case of RPGN
CPC 9: Case of NIDDM
CPC 10: Case of choriocarcinoma
APPENDIX:
Normal Values
WEIGHTS AND MEASUREMENTS OF NORMAL ORGANS
LABORATORY VALUES OF CLINICAL SIGNIFICANCE
INDEX
TOC
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