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Epidemiology
Etiology - Causes
Pathogenesis - Evolution
Morphology - Structural Changes
Clinical Significance – Functional Changes
Management
Complications
Prevention
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Four aspects of Pathology:
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Branches of clinical Pathology:
Histopathology Immunology
Anatomic
Cytopathology Chemical
Pathology
Haematology Pathology
Forensic Genetics
Pathology Toxicology
Microbiology
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Learning Pathology:
General Pathology
Common changes in tissues.
Systemic Pathology
Specific changes in organs.
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Techniques in Pathology:
Gross Pathology:
Light Microscopy:
Histopathology, Cytology, Autopsy
Histochemistry, Biochemical
Immunohistochemistry
Electron Microscopy
Cell Cultures, Medical Microbiology
Molecular Pathology
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HISTOTECHNOLOGY
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Surgical Specimen
Clinical Details
Adequate specimen
Proper Fixative
10% buffered
Formalin
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Gross Examination:
Description:
Specimen weight &
measurement (approx)
Consistency
Photo *
Cut section
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Taking Samples:
Edge of lesions.
Wall of cysts.
Include normal areas.
Avoid necrotic area.
Whole specimen if
small.
Direction, mark
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Inking the Margins*
To mark surgical
margin.
Spread of lesion
Malignancy
Adequacy of removal
Different colors to
identify margins
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Fixation:
After fixation is
Replacing aqueous
formalin with alcohol
in gradual sequence
(70, 95, 100%) to
make way for
paraffin.
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Clearing:
Removal of alcohol
with “xylene” that will
be miscible with the
embedding medium
(paraffin)
Impregnating with
paraffin.
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Embedding:
Paraffin block with
embedded tissue
consistency to cut
Paraffin blocks taken
for sectioning
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Tissue Processing:
Preservative
Provides stability
Protects from infection
Prevents autolysis
Permits sectioning and staining
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Sectioning:
Microtome
3-10 microns
Ribbon of sections
taken on hot water bath
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Picking up sections:
Floating
sections onto
slides
Common
“float” artefact
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Microscope slide preparation:
Taking the section onto
slide
Flat, no air bubbles, no
stretch or breaks.
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Automated Staining:
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Coverslipping:
Clearing - xylene
Thin glass coverslips to
protect the section
Using mounting media
(Eg. DPX, Resins,
Canada balsam etc.)
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Reporting:
Additional sections
Deeper / Thinner sect.
Special Stains/tech.
Reference..
Discussions with Clin.
Diagnosis
Report Typing
Despatch.
(>3-5 days)
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Cytopathology:
Cytopathology is study of cells in diagnosis
of disease.
Exfoliative & Non-Exfoliative - cytology.
Exfoliative: Cell samples are collected
from normally shedding tissues like
epithelium. Spatula or brush to enhances
collection.
Non-Exfoliative: Cells samples collected by
needles with suction pressure. (FNAC)
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PAP* Smear Normal:
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PAP Smear - Abnormal:
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Special Techniques:
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Light Microscopy
Kohler Illumination
Condenser
Objectives
2 to 4x - Low power
100x lens – Oil Imm.
Eye piece of 10x and
objective of 40x = 400
times magnification.
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Normal Stomach
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Normal Skin
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Normal Skeletal Muscle
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Normal Kidney
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Summary
1. Grossing
2. Fixation
3. Processing
4. Embedding
5. Sectioning
6. Staining
7. Mounting •
Some
Special
Techniques
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Frozen Sections:
Freezing acts as
embedding agent
by forming minute
ice crystals within
cells.
More rapid (5min),
Freezing Microtome
Liquid nitrogen.
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Immunohistochemistry
Antigen antibody
reaction
Marker Ab Tagged with
Sec. Antibody marker
Pri. Antibody Simple Dye
Enzyme (peroxidase)
Tissue Antigen
Fluorescent Dye
Radioactive Dye
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Melanoma +ve for HMB-45
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B cell Lymphoma – CD20
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Breast Cancer
Estrogen Receptor Antigen
Tamoxifen Sensitive
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Polarized Microscopy
Under Polarized light,
Some materials have
the property of
"birefringence" which
is the ability to pass
light in a particular
plane.
Eg. Crystals, fat,
fibers. Amyloid etc.
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Cardiac Amyloidosis
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Urine Oval Fat Bodies
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Fluorescent Microscopy
Property of materials
that causes them to
absorb light at a
shorter (UV)
wavelength, and to
emit light at a higher
(visible) wavelength
Auto-Fluorescence
Immuno-Fluorescence
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ANA – Diffuse Pattern
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ANA – Nucleolar Pattern
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Electron Microscopy
Electron beam instead of light.
Magnified images are typically from
1000X to 50,000X. (Light microscope is
10-1000x).
Gluteraldehyde fixative.
Glass knives.
Specimen is mounted on a metal grid.
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Membranous GN
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Minimal Change GN