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Pathology Practical Book
Harsh Mohan
SECTION ONE: TECHNIQUES IN PATHOLOGY
EXERCISE 1:
Microscopy
Light Microscope
STAND
BODY
OPTICAL SYSTEM
LIGHT/ILLUMINATION SYSTEM
MAGNIFICATION AND RESOLVING POWER OF LIGHT MICROSCOPE
HOW TO USE AND MAINTAIN A LIGHT MICROSCOPE?
NEWER APPLICATIONS OF LIGHT MICROSCOPE
Teaching Microscopes
Image Analysers and Morphometry
Telepathology
OTHER TYPES OF MICROSCOPY
DARK GROUND ILLUMINATION (DGI)
PHASE CONTRAST MICROSCOPY
POLARISING MICROSCOPE
FLUORESCENT MICROSCOPE
ELECTRON MICROSCOPE (EM)
Transmission Electron Microscopy (TEM)
Scanning Electron Microscopy (SEM)
EXERCISE 2:
Routine Histopathology Techniques and Staining
Fixation
TYPES OF FIXATIVES
1. Formalin
2. Glutaraldehyde
3. Bouin's Fluid (Picric acid)
4. Carnoy's Fixative (Alcohol)
5. Osmium Tetraoxide
Dehydration
Clearing
Impregnation
Tissue Processors
OPEN (HYDRAULIC) TISSUE PROCESSOR
CLOSED (VACUUM) TISSUE PROCESSOR
Embedding and Blocking
Section Cutting (Microtomy)
PROCEDURE FOR MICROTOMY
Routine Staining (H & E)
HAEMATOXYLIN
EOSIN
Procedure for Staining
Results
EXERCISE 3:
Frozen Section and its Staining
Frozen Section
APPLICATIONS
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
EXERCISE 4:
Special Stains and Immunohistochemistry
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
Immunohistochemical Stains
TECHNIQUE OF IHC
INTERPRETATION
APPLICATIONS OF IHC
EXERCISE 5:
Surgical Pathology Request Form
Design of the Request Form
Sample Request Form
SECTION TWO: GENERAL PATHOLOGY
EXERCISE 6:
Degenerations
Vacuolar Nephropathy
Hyaline Change in Leiomyoma
Myxoid Degeneration in Ganglion
EXERCISE 7:
Intracellular Accumulations
Fatty Change Liver
Melanin Pigment in Compound Naevus
Anthracotic Pigment in Lung
Brown Atrophy Heart
EXERCISE 8:
Amyloidosis
Amyloidosis Kidney
Amyloidosis Spleen
Amyloidosis Liver
EXERCISE 9:
Necrosis
Coagulative Necrosis (Infarct) Kidney
Liquefactive Necrosis (Infarct) Brain
Caseous Necrosis (Tuberculosis) Lymph Node
Enzymatic Fat Necrosis Pancreas
EXERCISE 10:
Gangrene and Pathologic Calcification
Wet Gangrene Bowel
Dry Gangrene Foot
Monckeberg's Arteriosclerosis
EXERCISE 11:
Derangements of Body Fluids
Pulmonary Oedema
CVC Lung
CVC Liver
CVC Spleen
EXERCISE 12:
Obstructive Circulatory Disturbances
Thrombus Artery
Pale Infarct Spleen
Haemorrhagic Infarct Lung
EXERCISE 13:
Types of Inflammation
Abscess Lung
Chronic Inflammatory Granulation Tissue
Tuberculous Lymphadenitis
EXERCISE 14:
Granulomatous Inflammation—Tuberculosis
Fibrocaseous Tuberculosis Lung
Tuberculosis Intestine
Miliary Tuberculosis Lung
Miliary Tuberculosis Spleen
EXERCISE 15:
Other Granulomatous Inflammations
Lepromatous Leprosy
Tuberculoid Leprosy
Sarcoidosis Lung
EXERCISE 16:
Fungal Infections
Oral Candidiasis
Madura Foot
Aspergillosis Lung
Rhinosporidiosis Nose
EXERCISE 17:
Actinomycosis and Parasitic Infections
Actinomycosis Skin
Cysticercosis Soft Tissue
Hydatid Cyst Liver
EXERCISE 18:
Adaptive Disorders of Growth
Testicular Atrophy
Cardiac Hypertrophy
Reactive Hyperplasia Lymph Node
Squamous Metaplasia Cervix
EXERCISE 19:
Primary Epithelial Tumours
Squamous Cell Papilloma
Squamous Cell Carcinoma
Malignant Melanoma
Basal Cell Carcinoma
EXERCISE 20:
Mesenchymal and Metastatic Tumours
Lipoma
Pleomorphic Rhabdomyosarcoma
Metastatic Carcinoma Lymph Node
Metastatic Sarcoma Lung
SECTION THREE: SYSTEMIC PATHOLOGY
EXERCISE 21:
Atherosclerosis and Vascular Tumours
Atheroma Aorta
Capillary Haemangioma Skin
Cavernous Haemangioma Liver
Lymphangioma Tongue
EXERCISE 22:
Diseases of the Heart
Bacterial Endocarditis
Healed Myocardial Infarct
Chronic Ischaemic Heart Disease
Fibrinopurulent Pericarditis
EXERCISE 23:
Pneumonias
Lobar Pneumonia— Acute Congestion Stage
Lobar Pneumonia— Red Hepatisation Stage
Lobar Pneumonia— Grey Hepatisation Stage
Bronchopneumonia
EXERCISE 24:
Chronic Obstructive Pulmonary Diseases and Lung Cancer
Emphysema
Bronchiectasis
Squamous Cell Carcinoma Lung
Small Cell Carcinoma Lung
EXERCISE 25:
Diseases of the Jaws, Salivary Glands and Stomach
Ameloblastoma
Pleomorphic Adenoma
Peptic Ulcer
Adenocarcinoma Stomach
EXERCISE 26:
Diseases of Appendix and Large Intestine
Acute Appendicitis
Ulcerative Colitis
Juvenile Polyp Rectum
Colorectal Mucinous Adenocarcinoma
EXERCISE 27:
Hepatitis—Viral, Alcoholic, Fulminant
Acute Viral Hepatitis
Alcoholic Hepatitis
Fulminant Hepatitis
EXERCISE 28:
Cirrhosis, Hepatocellular Carcinoma, Gallbladder Diseases
Cirrhosis Liver
Hepatocellular Carcinoma
Chronic Cholecystitis with Cholelithiasis
Carcinoma Gallbladder
EXERCISE 29:
Primary Glomerulonephritis and Tubulointerstitial Disease
Acute Glomerulonephritis
Rapidly Progressive Glomerulonephritis (RPGN)
Chronic Glomerulonephritis
Chronic Pyelonephritis
EXERCISE 30:
Diabetic Nephropathy and Tumours of Renal System
Diabetic Nephropathy
Renal Cell Carcinoma
Wilms' Tumour
Transitional Cell (Urothelial) Carcinoma
EXERCISE 31:
Testicular Tumours and Diseases of Prostate
Seminoma Testis
Nodular Hyperplasia Prostate
Adenocarcinoma Prostate
EXERCISE 32:
Diseases of Endometrium, Placenta, Cervix
Simple (Cystic Glandular) Hyperplasia
Hydatidiform Mole
Invasive Cervical Cancer
EXERCISE 33:
Ovarian Tumours
Serous Ovarian Tumours
SEROUS CYSTADENOMA OVARY
PAPILLARY SEROUS CYSTADENOCARCINOMA OVARY
Mucinous Ovarian Tumours
MUCINOUS CYSTADENOMA OVARY
Benign Cystic Teratoma Ovary
EXERCISE 34:
Diseases of Breast
Fibrocystic Change
Fibroadenoma
Infiltrating Duct Carcinoma-NOS
EXERCISE 35:
Diseases of Thyroid
Hashimoto's Thyroiditis
Nodular Goitre
Follicular Adenoma
Papillary Carcinoma
EXERCISE 36:
Osteomyelitis and Benign Bone Tumours
Pyogenic Osteomyelitis
Tuberculous Osteomyelitis
Osteochondroma
Osteoclastoma
EXERCISE 37:
Malignant Bone Tumours
Ewing's Sarcoma
Osteosarcoma
Chondrosarcoma
EXERCISE 38:
Meningitis and CNS Tumours
Acute Pyogenic Meningitis
Meningioma
Schwannoma
Astrocytoma
SECTION FOUR: CYTOPATHOLOGY
EXERCISE 39:
Basic Cytopathologic Techniques and their Applications
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
Procedure for FNA
Method
RADIOLOGICAL IMAGING AIDS FOR FNA
ADVANTAGES OF FNA OVER SURGICAL BIOPSY
Imprint Cytology
APPLICATIONS OF IMPRINT CYTOLOGY
ADVANTAGES OF IMPRINT CYTOLOGY
EXERCISE 40:
Interpretation of Pap Smears and Fluid Cytology
Inflammatory Pap Smear
Pap Smear in Carcinoma Cervix
Fluid Cytology for Malignant Cells
EXERCISE 41:
Interpretation of FNA Smears
FNA in Tuberculous Lymphadenitis
FNA in Fibroadenoma Breast
FNA in Duct Carcinoma Breast
SECTION FIVE: CLINICAL PATHOLOGY
EXERCISE 42:
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
2. Refractometer
3. Reagent Strip Method
Significance of Specific Gravity
C. Chemical Examination
TESTS FOR PROTEINURIA
TEST FOR GLUCOSURIA
TESTS FOR KETONURIA
1. Rothera's Test
2. Gerhardt's Test
3. Reagent Strip Test
TEST FOR BILE DERIVATIVES IN URINE
Cause for Bile Salts in Urine
1. Fouchet's Test
2. Foam Test
3. Reagent Strip Test
TESTS FOR BLOOD IN URINE
Automated Urinalysis
EXERCISE 43:
Urine Examination II: Microscopy
A. Collection of Sample
B. Preparation of Sediment
C. Examination of Sediment
1. CELLS IN URINE
RBCs
WBCs
Epithelial Cells
2. CASTS IN URINE
i. Hyaline Cast
ii. Red Cell Cast
iii. Leucocyte Cast
iv. Granular Casts
v. Waxy Casts
vi. Fatty Cast
vii. Epithelial Cast
viii. Pigment Cast
3. CRYSTALS IN URINE
4. MISCELLANEOUS STRUCTURES IN URINE
D. Automation in Urine Analysis
URINE STRIP ANALYSERS
FLOW CYTOMETRY
EXERCISE 44:
Semen Analysis
Sample Collection
Gross Examination
Microscopic Examination
MOTILITY
COUNT
MORPHOLOGY
Chemical Examination
FRUCTOSE TEST
ACID PHOSPHATASE TEST
Immunological Assays
Microbiological Assays
Sperm Function Tests
Semen Cryopreservation
EXERCISE 45:
Examination of CSF
Normal Composition of CSF
Specimen Collection
Microscopic Examination
TOTAL LEUCOCYTE COUNT
Manual Method
Automated Method
DIFFERENTIAL LEUCOCYTE COUNT
Chemical Examination
Microbiological Examination
Immunological Examination
SECTION SIX: HAEMATOLOGY
EXERCISE 46:
Types of Blood Samples, Anticoagulants and Blood Collection
Types of Blood Samples
WHOLE BLOOD
Uses
SERUM
Uses
PLASMA
Uses
Anticoagulants
1. EDTA
Uses
2. SODIUM CITRATE
Uses
3. DOUBLE OXALATE
Uses
4. HEPARIN
Uses
5. Sodium fluoride
Blood Collection
EXERCISE 47:
Haemoglobin Estimation
Methods for Estimation of Haemoglobin
CYANMETHAEMOGLOBIN METHOD
OXYHAEMOGLOBIN METHOD
ELECTRONIC COUNTER METHOD
Disadvantage
DIRECT READING ELECTRONIC HAEMOGLOBINOMETERS
Disadvantage
SAHLI'S METHOD
OTHER METHODS
NORMAL VALUES OF HAEMOGLOBIN
ERRORS IN HAEMOGLOBINOMETRY
Quality Control in Haemoglobin Estimation
EXERCISE 48:
Counting of Blood Cells
WBC Count
METHODS
Visual Haemacytometer Method
Electronic Method
NORMAL RANGE FOR WBC COUNT
CAUSES OF ABNORMAL LEUCOCYTE COUNT
RBC Count
METHODS FOR RBC COUNTS
Visual Haemacytometer Method
Calculations
Electronic Method
NORMAL RANGE FOR RBC COUNT
CAUSE OF DECREASED RBC COUNT
CAUSE OF INCREASED RBC COUNT
Platelet Count
METHODS FOR COUNTING PLATELETS
Visual Method
Rough Visual Method for Platelet Counting
Electronic Method
NORMAL PLATELET COUNT
CONDITIONS CAUSING THROMBOCYTOPENIA
CONDITIONS CAUSING THROMBOCYTOSIS
Absolute Eosinophil Count (AEC)
METHODS
Improved Neubauer Chamber Method
Calculations
Fuchs-Rosenthal Counting Chamber Method
NORMAL RANGE FOR AEC
CAUSES OF ABNORMAL AEC
EXERCISE 49:
Reticulocyte Count
Reticulocytes
Methods for Counting of Reticulocytes
VISUAL METHOD
Counting of Reticulocytes
AUTOMATED METHOD
NORMAL VALUES
ABNORMAL COUNTS
EXERCISE 50:
Preparation and Staining of Peripheral Blood Film
Thin Blood Film
SLIDE METHOD
COVER GLASS METHOD
SPIN METHOD
Thick Blood Film
Stains for Blood Film
STAINING OF THIN BLOOD FILM
Leishman Stain
Giemsa Stain
STAINING OF THICK SMEAR
AUTOSTAINERS
PRECAUTIONS IN STAINING OF PBF
EXERCISE 51:
Differential Leucocyte Count
Examination of PBF for DLC
Morphologic Identification of Mature Leucocytes
POLYMORPH (NEUTROPHIL)
LYMPHOCYTE
MONOCYTE
EOSINOPHIL
BASOPHIL
Methods of DLC
VISUAL COUNTING
AUTOMATED COUNTING
Pathologic Variations in DLC
NEUTROPHILS
LYMPHOCYTES
MONOCYTES
EOSINOPHILS
BASOPHIL
EXERCISE 52:
ESR, PCV (Haematocrit) and Absolute Values
Erythrocyte Sedimentation Rate
PRINCIPLE
MECHANISM OF ESR
PHASES IN ESR
METHODS OF ESR
1. WESTERGREN'S METHOD
2. WINTROBE'S METHOD
3. MICRO ESR METHOD
4. AUTOMATED ESR METHOD
CLINICAL SIGNIFICANCE OF ESR
Diagnostic Uses
Monitoring Prognosis of Diseases
Packed Cell Volume or Haematocrit
METHODS FOR ESTIMATION OF PCV
1. MACRO (WINTROBE'S) METHOD
2. MICROHAEMATOCRIT METHOD
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 53:
Screening Tests for Haemostasis
Bleeding Time
METHODS FOR BLEEDING TIME
1. Finger Tip Method
2. Duke's Method
3. Ivy's Method
Clinical Application of Bleeding Time
Clotting Time
METHODS FOR CLOTTING TIME
1. Capillary Tube Method
2. Lee and White Method
Clinical Applications of Clotting Time
EXERCISE 54:
Blood Grouping and Cross-matching
ABO System
INCIDENCE OF ABO BLOOD GROUPS IN INDIA
METHODS FOR ABO GROUPING
1. SLIDE METHOD
2. TUBE METHOD
Cell Grouping (Fig. 54.2,A)
Serum Grouping (Fig. 54.2,B)
Advantages of Tube Method Over Slide Method
NOTE
Rhesus (Rh) System
METHODS FOR RH GROUPING
1. SLIDE METHOD
2. TUBE METHOD
NOTE
Cross-matching
PROCEDURE
INTERPRETATION
Importance of Blood Grouping
EXERCISE 55:
Peripheral Smear Examination in Anaemias
Plan of Investigation for Anaemia
HAEMOGLOBIN ESTIMATION
PERIPHERAL BLOOD FILM 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
PBF in Microcytic Hypochromic Anaemia (Iron Deficiency Anaemia)
PBF in Macrocytic Anaemia (Megaloblastic Anaemia)
PBF in Haemolytic Anaemia (Thalassaemia)
EXERCISE 56:
Bone Marrow Examination
Bone Marrow Aspiration
Trephine Biopsy
EXERCISE 57:
Haemoparasites
PBF in Malarial Parasite
PBF in Filariasis
Bone Marrow in Leishmaniasis
EXERCISE 58:
Haematologic Malignancies: Leukaemias
Common Classification of Leukaemias-Lymphomas
Acute Leukaemia
Chronic Myeloid Leukaemia
Chronic Lymphocytic Leukaemia
EXERCISE 59:
Haematologic Malignancies: Lymphomas
Non-Hodgkin's Lymphoma (NHL)
SLL-CLL
FOLLICULAR LYMPHOMA
Hodgkin's Disease (HD)
HD-NODULAR SCLEROSIS TYPE
HD-MIXED CELLULARITY TYPE
SECTION SEVEN: AUTOPSY PATHOLOGY
EXERCISE 60:
Autopsy Protocol—Request and Report
Autopsy Protocol
A. PROSECTION
B. GROSS EXAMINATION OF ORGANS
C. AUTOPSY REPORT
EXERCISE 61:
CD on Clinicopathological Conferences and Museum Review
Clinicopathological Conference (CPC)
CD and its Contents
SECTION I: TEN CPCs
CPC 1: Case of secondary systemic amyloidosis
CPC 2: Case of septic shock with DIC
CPC 3: Case of HIV infection with miliary tuberculosis with tuberculous meningitis (TBM)
CPC 4: Case of rheumatic heart disease (RHD) with infective endocarditis
CPC 5: Case of alcoholic cirrhosis with portal hypertension
CPC 6: Case of bronchogenic carcinoma
CPC 8: Case of rapidly progressive glomerulonephritis (RPGN)
CPC 9: Case of type II diabetes mellitus with complications
CPC 10: Case of gestational choriocarcinoma
SECTION II: QUICK REVIEW OF PATHOLOGY MUSEUM
SECTION EIGHT: APPENDICES
APPENDIX I:
Common Instruments in Pathology
A. Haematology (1-20)
1. ANTISERA FOR BLOOD GROUPING
2. BONE MARROW ASPIRATION (SALAH'S) NEEDLE
3. DLC COUNTER (MANUAL)
4. EHRICH'S EYEPIECE
5. GRADUATED PIPETTE (1 ML)
6. LANCET
7. NEUBAUER'S (IMPROVED) COUNTING CHAMBER (HAEMACYTOMETER)
8. PASTEUR PIPETTE
9. RBC PIPETTE
10. SAHLI'S COLOUR COMPARATOR
11. SAHLI'S HAEMOGLOBIN PIPETTE
12. SAHLI'S HAEMOGLOBIN TUBE
13. STOP WATCH
14. THERMOMETER FOR WATERBATH
15. TREPHINE BIOPSY (JAMSHIDI'S) NEEDLE
16. VACUTAINERS WITH VACUPUNCTURE NEEDLE
17. WBC PIPETTE
18. WESTERGREN'S PIPETTE
19. WINTROBE'S STAND
20. WINTROBE'S (HAEMATOCRIT) TUBE
B. Clinical Pathology (21-25)
21. DIPSTIX FOR URINE
22. ESBACH'S ALBUMINOMETER
23. LUMBAR PUNCTURE NEEDLE
24. URINOMETER
25. VIM SILVERMAN'S NEEDLE
C. Histopathology (26-31)
26. BLOCK MOULDS (METAL), LEUCKHART'S (L)
27. BLOCK MOULDS (PLASTIC)
28. MICROTOMY BLADES, HIGH AND LOW PROFILE
29. MICROTOMY KNIFE, PLAIN WEDGE TYPE
30. SLIDE CARRIER
31. TISSUE CAPSULES (STAINLESS STEEL)
D. Cytopathology (32-34)
32. AYRE'S SPATULA
33. COPLIN JAR
34. FRANZEN HANDLE
APPENDIX II:
Common Similes in Pathology
APPENDIX III:
Normal Values
Weights and Measurements of Normal Organs
Laboratory Values of Clinical Significance
INDEX
TOC
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