Introduction to Anatomical

Pathology

Krisna Murti

Anatomical Pathology, Faculty of Medicine, Sriwijaya

University
 Historical Periods of Pathology
1. Anatomical (before XIX century)
J. Morgagni (1685-1777)
K. von Rokitansky (1804-1878) performed nearly 30,000
autopsies, wrote an outstanding monograph on diseases
of arteries and congenital heart defects

2. Microscopic (XIX century - mid-XX

century)
R. Virchow (1821-1902) father of cellular pathology

3.Ultramicroscopic (mid-XX century)

4.Modern - pathology of a living person

 Definition

Pathology (Anatomical Pathology / Pathologic

Anatomy)

is fundamental biomedical science that studies

the structural basis of pathological processes of
human disease
Anatomical Pathology is a medical laboratory
specialty, whose objective is the diagnosis of
diseases through the study of biological material
obtained from organs or tissues and that may
consist of cells or fluids.
 The Scope of Pathology

1. The core of pathology:

The four aspects of a disease process that form the core of pathology:
Epidemiology
(1) Etiology: causes of the disease
(2) Pathogenesis: the evolution/mechanisms of its development
(3) Morphologic changes: the structural alteration induced in the cells and
organs
of the body
(4) Clinical significance: the functional consequences of the morphologic
changes

Management
Complications
Prevention
2. Classification:
Autopsy
(1) Human pathology
Biopsy
Cytology
(2) Experimental pathology

3. Position:
Its a bridging discipline involving both
basic science and
clinical practice
4. Text of Pathology:

(1) General pathology:

concerned with the basic reaction of cells and tissues to
abnormal
stimuli that underlie all diseases

(2) Systemic pathology :

describe the specific responses of specialized organs and
tissues
to defined stimuli
 Techniques of Pathology

1. Human pathology
(1) Autopsy
(2) Biopsy: surgical or diagnostic pathology
(3) Cytology: smear, fine needle aspiration

2. Experimental pathology
(1) Animal experiment: animal model
(2) Tissue and cell culture
Morphological Alteration
Structural alteration of cells or tissuesusually
specific for particular diseases

Clinical symptoms: alteration of function due to

morphological changes

Morphological alterations of tissues / organs

cause malfunction of the organs and then raise
clinical symptoms
Observation and New Technique of
Morphology

Gross appearance:
size, shape
weight
color
consistency
surface
edge of section
Histologic and Cytologic Observation

most common and basic formalin fixed

HE (hematoxylin and eosin) stained

Hemangioma of ventrical wall

 Methods of Pathological Anatomy

Biopsy
Removal of tissue from a living subject to determine
morphological changes;

Autopsy
Post-mortem examination of a corpse to determine the
cause and manner of death and to evaluate any
disease or injury that may be present

Experiment
Modelling of pathological process on animals and
subsequent post-mortem examination
 Types of Biopsies

During surgery
Puncture (needle
biopsy)
Aspiration
Excisional
Incisional
Scrape
 Anatomical Pathology

Histopathology
Cytology
Gynecology: papsmear,
Non gynecology: FNAC, sputum, pleural effusion,
Transthoracal biopsy (TTB), transthoracal needle
aspiration (TTNA)
Vriescope (VC)
Histochemistry
Imunohistochemistry
Molecular
 Levels of Study

Organismal
Organ
Tissue
Cellular
Ultrastructural
Molecular
 Histopathology

Macroscopic/gross

Direct examination by eyes or by touching

Organs :
Sizes : enlargement / smaller
Consistency : soft, hard, solid, fragile
Color : pale, yellow, brownish
 Microscopy

Using microscope

Helped by staining for histochemistry or

immunohistochemistry to show specific
chemical contains in cells or tissues

Observe structural alterations in cells /

tissues due to particular diseases
Histopathological
Materials

Biopsy: excise, endoscopy, cystoscopy

Operation : appendices, ovary, breasts

Extirpation : Lymph nodes

Curettage : specific for abnormalities in

endometrium, cervix and utery
Eg. abortus, mola hidatidosa, hormonal
abnormality, malignancies
Procedures

a. Formalin 10% buffer

b. The whole tissues rinse in fixation (volume of
formalin is 10 X tissues volume)
c. If the tissues are too large lamellation every
1 cm to ensure fixation enters the tissues
d. Processingmachines
 Processing

Removal of alcohol
with xylene that will
be miscible with the
embedding medium
(paraffin)

Impregnating with
paraffin.
Haematoxylin-Eosin Staining
(Routine)

Machine

Manual
Cytology

Cytos (cells) and logos (knowledge)

Exfoliative cytology: knowledge of cells

important for early diagnosis of diseases /
malignancies

Spontan exfoliation occur due to mature cells

changed by younger cells the normal
behavior of superficial tissues / organs
A. Structure of cells
A cell is an important structure

Different forms depending of the functions

The cell from different tissues / organs has specific

behavior

B. Structure of tissues
Tissues consist of collection of cells
Pap smearmany types of cells originated from
different tissue types of female genital tracts
Cytology
Cytology: to observe and examine structural alteration of every
found cell

To detect malignancies: pap smear/pap test/ papanicolau

test/cervical smear early detection of cervical cancer

Genetic disorders

Hormonal disorders

a. Gynecologic: swab of lateral vaginal wall, cervix and

endometrium

b. Non gynecologic: sputum (lung), ascites, pleural

effusion
Gynecologic

1. Hormonal evaluation
2. Early detection of cancer
3. Inflammation detection

Procedures:
- Lateral wall of vagina (1/3 inner wall)
- Cervix
- Endometrium
Pap Smear/Cervical Smear
The examination to detect abnormalities of cervix in
particular cervical cancer
Taken from fornix posterior/squamo-columnar junction
fixation : alcohol 95%
Staining : Papanicolau

Representative if Endocervical cells are present

Not representative/inadequate if :
Too little
Swab is too thick
Too much blood
Inadequate fixation
Endocervical cells are not present
Pap Smear
Non Gynekologic

1. Respiratory tracts
- sputum
- Bronchial washing
- TTB (transthoracic biopsy)
- TTNA
2. Digestive tracts endoscopy for stomach
3. Urinary tracts urine
4. Pleural cavity, pericard, sinovial, abdomen (ascites),
cerebrospinal fluid
Fine Needle Aspiration Cytology (FNAC)
Staining : PAP, HE, MGG
Aplications : superfisial and deep organs
Superfisial : lien, breasts, thyroid, peripheral lung,
pleura, peritoneal, sinovial, tumor
Deep organs: deep lung lesion, liver, kidenys, prostate

Superficial
Needle Biopsy

Deep
Procedures

Pap smear Fixation: alcohol 95%

The swab as soon as possible rinse into alcohol 95%
for minimal 30 min

Urine/fluid from body cavities (pleural effusion,

ascites)
- Can be sent as swab of sediment after centrifugation
- Fixation: alcohol 95%
- Or urine/pleural effusion/ascites about 100-200 cc fix
with alcohol 50% aa
3. FNAC (fine needle aspiration cytology)
Dry with no fixation: dry the swab on the air without
fixation then send
to the lab

Wet Fixation: swab rinses into alcohol 95% for

minimal 30 min then send to the lab
 Frozen Section/Vries Coupe/VC

Examination of
histopathology
while the patient
is in the
operation room

To ensure the
malignancy is
present or not
then therapy
Procedures in sending the
specimens
1. Fill in forms completely
2. Send the specimens in adequate fixation
(formalin 10% buffer)
Histochemistry and
Cytochemistry

PAS staining
Autopsy

Clinical autopsy is part of pathologists duties

Identify new diseases
Identify etiologies of death
Examination of the correctness of the
diagnosis and treatment
Establish the cause of death
Research
Teaching students and physicians
Judicial Autopsy (Autopsi
kehakiman)
Forensic

Criminal cases
Legal Authority of Autopsy
Autopsy is performed mandatory:
Suspicion of violent death
Death less then 1 day after admission to
hospital
Death during surgery, diagnostic
manipulations and/or
anesthesia
Death from infection
Suspicion of overdose or drug intolerance
Pregnant women, women in/after childbirth
Children under 1 year
 Terminologies

Alteration of epithelia in malignancy

Variation in nuclear size and forms
(pleomorphism)
Alteration in nuclear chromatin:
hyperchromatic, vesicular
Increase NC ratio
Nucleoli can be seen

: Metaplasia
Alteration of a mature cell type to another
type
Eg: changes of columnar epithelial-
Cont

Dyplasia: alteration of mature cell type to

disorientation

Undifferentiated cell: not differentiate not growth to

specialized types
Cont
Hyperchromatism: an increase in the histological
staining, usually in the nucleus

Hyperplasia: an increase in the amount of

organic tissue that results from cell proliferation

Hypertrophy: An enlargement of organs due to the

increase of cell size

Physiology: athletes muscles, pregnant uteri

Pathology: hypertensioncardiac enlargement

 Cont
Karyolysis: The complete dissolution of the chromatin of a
dying cell due to the enzymatic degradation by
endonucleases
Karyorrhexis: the destructive fragmentation of the nucleus
of a dying cell whereby its chromatin is distributed
irregularly throughout the cytoplasm
Carcinoma: Malignant neoplasm originated from epithelial
Pyknosis : the irreversible condensation of chromatin in the
nucleus of cell undergoing necrosis or apoptosis
Polymorphism : occur when two or more clearly different
phenotypes exist in the same population of a species the
occurrence of more than one form or morph or multiple
alleles in a single gene
Pleomorphism : Occurring in various distinct forms. In
 Flow Cytometry (FCM)
1. One kind of cellsquantitative analysis
2. DNA ploidy analysis
3. Protein nucleus acidquantitative analysis

4. Selection of collection of cells

Image analysis (IA)
Nuclei: diameter; circumference; area; volume;
morphology

Laser scanning confocal microscope (LSCM)

Aliving cellobservation in situ or development
or quantitative
Modern Methods in Morphology

Immunohistochemistry (IHC)

Electron microscopy

In situ hybridization (ISH)

Polymerase chain reaction

(PCR)
 Immunohistochemical techniques
Immunohistochemistry
It is based on specific interaction of tissue
and cellular antigens with a specially derived
antibodies bearing the different labels.
Immunohistochemistry (IHC)

Immunofluorescence (IF)

Opportunities IHC

Determination of cells belonging to a

particular tissue;
Identification of individual products (e. g.
abnormal
proteins), routes of cellular and
intercellular signals,
synthesis of certain proteins, glyco-and
lipoproteins
 Immunohistochemistry

1. Ag-Ab specific reaction

2. Applications
(a) Location analysis
cytokeratincell membrane
(b) Clinical diagnosis and distinguishing
diagnosis of tumor histogenesis
Immunofluorescence (IF)
 Ultrastructure Observation
Used to study the details of cell structure, detection of viruses,
bacteria, immune complex deposits

Examples of use:

Oncology - Birbeck granules in histiocytosis X

Oncology - Z-discs in rhabdomyosarcoma cells
Nephrology - diagnosis of glomerulonephritis

TEM (transmitting electron microscope) SEM (scanning electron microscope)

 In Situ Hybridization (ISH)
In situ hybridization (ISH)
Method for localizing / detection of specific sequences
(of DNA or RNA in situ / directly in tissue specimens) in
morphologically preserved tissue sections or cell
preparations by hybridizing the complementary strand of
a nucleotide probe against the sequence of interest.

It is based on principle of complementary interaction of

DNA or RNA in specimen with labelled nucleotide
sequence (probe)

If nucleic acids are preserved in a histological specimen,

then it can be
Is used for:
detected by using a complementary probe

Detection of viral genomes

Detection of mutant genes
Detection of active protein synthesis
(unlike IHC which allows to determine the
presence of a protein)
IHC data validation
 Polymerase Chain Reaction (PCR)

Method for detection of specific sequences of

DNA or RNA in any biological sample

PCR is in vitro amplification (i. e. increase in the

number of copies) of nucleic acids triggered by
synthetic oligonucleotide primers

Differences from the in situ hybridization:

Because of amplification it is more

sensitive (about 1 million times)

Usually not combined with morphology

Molecular Biology Technique

1. Polymerase chain reaction (PCR)

2. DNA sequencing
3. Biochip technique
(1) Gene chip (DNA chip)
(2) Protein chip (protein microarray)
(3) Tissue chip (tissue microarray)
Thank you