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Dr. Roman | June 17, 2015 | PATHOLOGY

SURGICAL PATHOLOGY: An Introduction

Italicized from the lecture of Dr. Roman SURGICAL PATHOLOGY REPORT

th
Blue Text from Surgical Pathology 10 Ed. Should contain all relevant gross and microscopic
features of a case
PATHOLOGY Should be:
ANATOMIC PATHOLOGY Prompt
Autopsy Accurate
Surgical Pathology Brief
Cytology Avoids unnecessary histologic jargon
Forensic Pathology
CLINICAL PATHOLOGY 5 Major Fields:
Hematology 1. History
Clinical Chemistry Clinical data known to the pathologist
Blood Banking Name, age, sex, symptoms, surgical findings,
Immunology/Serology type of surgery
Clinical Microscopy Optional; includes the demographics of the
Microbiology patient
Contains the essential clinical data known to
WHO IS SURGICAL Pathologist? the pathologist at the time he dictates a
Must be a licensed physician. description of the gross specimen(s).
Must undergo training in an accredited hospital Gives the reader of the report, whether a
by the Philippine Society of Pathologist clinician or another pathologist, an immediate
Must pass the Specialty Board Examination given orientation to the nature of the problem that
by the PSP Board of Pathologists. led to that particular operation.
Either a Diplomate or a Fellow 2. Gross
Pathology Residency = 4 years Gross description
Pathologist the one who reads the Precise and thorough, (with picture is better)
biopsies. Fresh or fixed specimen, previously opened
or intact, any landmark (ex. Suture)
Surgical Pathologist Size, color and location of all lesions should
Closely affiliated with all surgical specialties, be recorded.
internal medicine, dermatology, neurology, Use metric system cm used not inches
radiology etc. Exact measurement is better than comparing
He needs to understand the clinicians needs and with objects (e.g. size of an egg which
respond to them accordingly. egg?) give 3D measurements
He needs to tell the clinician, not only whether a It is advisable to give specific dimensions and
tumor is benign or malignant but also descriptions rather than to provide
Extent of the disease comparisons with common objects such as
Grade of malignancy fruits or other vegetables.
Adequacy of surgery Weight is important in many tissues, like
Other pertinent information, e.g. thyroid, prostate normal weight of the
Prognosis prostate = 20g
It is important to be accurate, factual, and
Subspecialties noncommittal in the gross description,
Hematopathology avoiding subjective interpretations as much
Renal Pathology as possible.
Neuropathology Use standard color, like red, brown, amber,
Dermatopathology green rather than blood-stained, pus or milky.
Forensic Pathology using blood-stained, pus or milky somehow

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 imparts the diagnosis rather than describe
the specimen
Concluded by stating whether the entire
specimen was submitted for tissue
processing or sampling was done.
3. Microscopic Description
Optional
Short and straight to the point
Histologic samples are kept by the lab
Usually not written or described
4. Diagnosis
Organ, specific site, operation morphologic
Diagnosis
E.g. Bone, femur, biopsy Osteosarcoma.
5. Note or Remarks
Differential diagnosis, reasons for
interpretation, prognostication and
therapeutic consideration
Helps the clinician make his/her diagnosis
and plan of care
One of the most important means of
clinicopathologic correlation
REMEMBER!
no technologic advancement can replace the
time-honored practice of two medical specialist
discussing how best to treat a patient.
SLIDE REVIEW
Slide can be evaluated by several pathologists or
by the same pathologist at a different time.
Slides and paraffin blocks are stored indefinitely.
Usual practice is to review outside slides of
patient who is referred to his institution prior to
therapy
Consultation with other pathologists in unusual,
difficult and controversial cases is now a popular
practice
Slides sent to different professionals for second
opinions.
LIMITATIONS OF HISTOLOGIC DIAGNOSIS
Pathologists are physicians and human beings.
They have as great a capacity for error and
susceptibility to subjective distractions as other
practitioners of the art of medicine.
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Incomplete communication between clinician and
pathologist may make diagnosis difficult or
impossible.
If diagnosis is not compatible with clinical
diagnosis, send slide to other
pathologists for second opinion
Sampling of tissue need enough samples
which vary depending on the pathologist
Best biopsy is a cleanly excised, uncrushed
wedge that includes a junction between normal
and neoplastic tissue.
Adequacy of sample - Adequate volume of tissue
permits a choice of fixatives, histochemical
studies, bioassay or tissue culture.
Fixation of sample fix sample at once
Upon removal of the breast tissue, it takes 20
minutes before receptors autolyse secondary
to anoxia. If the sample was fixed after > 20
minutes, the test would yield a false negative
result.
BIOPSIES
Incisional Biopsies
Only a portion of the lesion is sampled.
Strictly of diagnostic nature.
Involves the removal of portion/s of a big
mass
Excisional Biopsies
Entire lesion is removed usually with a rim of
normal tissue.
Both diagnostic and therapeutic in some
cases.
Factors to consider:
Size of the lesion
Depth of lesion
Wide Excision for suspected malignant
lesion
Fine Needle Aspiration Biopsy current trend in tissue
sampling.
- Accurate results of cytology exam
- Usually done with lung and liver biopsy
General Rules for Biopsy Procedure:
Larger lesions should be sampled more
variability in pattern may exist and the diagnostic
areas may be present only focally.
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 In ulcerated lesion, take from the periphery that
includes both normal and diseased tissues -
biopsy of the central ulcerated area may show
only necrosis and inflammation
Biopsy should be deep enough to show
relationship between the tumor and stroma -
Epithelia involved by carcinoma have a tendency
to detach from the underlying stroma. This should
be avoided whenever possible by careful
handling of the tissue.
Deep seated lesion can show peripheral tissue
reaction, e.g. inflammation, fibrosis, calcification
If the biopsy is too peripheral, this may only
reflect peripheral tissue reaction. Similarly, in a
mass of lymph nodes, a deep-seated node may
show involvement by a malignant tumor, whereas
a superficial node may show only nonspecific
hyperplasia.
When several fragments of tissues are obtained,
submit everything - Sometimes the smaller or
grossly less impressive fragment is the only one
that contains the diagnostic elements.
Get at the junction between normal tissue and the
ulcer. Obtaining sample in the middle will only yield
necrotic tissue.
Crushing or squeezing with forceps should be
avoided. increases artifacts making
interpretation difficult
Always place adequate amount of fixative
10% buffered neutral formalin
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Volume of fixative is 10x that of tissue
(1:10 ratio)
Consider the need for special studies, e.g. touch
preparations, electron microscopy, cytogenetics,
molecular pathology, flow cytometry, etc.
INTRAOPERATIVE CONSULTATION (FROZEN
SECTION)
Required experience, knowledge of clinical
medicine and pathology, capacity to make quick
decisions under pressure, good judgment and
attitude that is conservative but no excessively
so.
Cryostat or Freezing microtome.
A simple question:
Will the result of the frozen section
examination influence in any way the
surgical procedure?
If the answer is no, the procedure is not
indicated.
E.g. A patient with Hirschsprungs Disease
undergo Frozen Section intraop to determine the
area where there are already ganglion cells
present before performing a pull-through surgery
Three legitimate purpose:
To establish the presence and nature of
a lesion.
To determine adequacy of surgical
margins
To establish whether the tissue obtain
contains diagnosable material or whether
additional sampling is indicated
Diagnosis usually given within 20 minutes
especially with breast specimens
At the time of a frozen section, the diagnosis
given verbally to the surgeon should be
transcribed verbatim in an appropriate form and a
copy of such form incorporated immediately into
the chart.
Another copy should remain in the laboratory and
be filed with the frozen section slides.
Overall accuracy is pretty good:
CAP sponsored review show a
concordance of 98.58%
Causes of discrepancies between frozen sections
and paraffin sections:
Misinterpretation of the original frozen
section
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 Absence of diagnostic tissue in the
material frozen but present in tissue not
sampled.
Absence of diagnostic tissue in the
material frozen but present in permanent
section
A correlation exists between the accuracy of the
procedure and both the tissue type and the
nature of the pathologic process.
The real aim of the frozen section procedure is to
influence the course of the operation.
The pathologist should be thoroughly briefed on
the patient's clinical history: ideally the surgeon
and the pathologist should have discussed the
case beforehand. The pathologist should be
prepared to advise the surgeon as to the best
area to biopsy.
Freezing the tissue in isopentane (methylbutane)
cooled with liquid nitrogen or with an electronic
device saves valuable time and results in fewer
artifacts than when the tissue is frozen on the
cryostat stage.
Parts:
Gross examination determine if benign
or malignant from gross features
Touch imprint touch the slide on the
surface of the tumor to get samples
Samples are examined
cytologically
Microscopic examination
DIAGNOSTIC CYTOLOGY
In the hands of a well-trained, experienced
individuals offers an extremely high degree of
reliability
A positive cytologic diagnosis of malignancy
made under these circumstances should be given
the same weight as one obtained from a surgical
biopsy.
Terminology used should be the same as those
used for biopsies e.g. HSIL rather than Moderate
Dysplastic cells.
In most organs, the cytology diagnosis should be
followed by conventional biopsy before treatment
is carried out e.g. cervical cancer.
Pap smear showing severe dysplasia is
followed by cervical biopsy to identify
layer involvement
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FNAB breast, thyroid, salivary glands and lung.
It is generally carried out with a fine
needle (OD 0.60.9 mm), sometimes
under image guidance.
Inexpensive, safe, quick, and when
performed by experienced workers
quite accurate.
It can also induce artifacts of various
types in the tissues, which the pathologist
should be cognizant of in order to avoid
misinterpretation.
Many of the special stains that are routinely used
for tissue sections can also be very useful for the
evaluation of cytologic material. This includes
stains for glycogen, melanin, fat, and mucin.
Cytologic material is also well suited for
examination with immunocytochemical,
ultrastructural, flow cytometric, cytogenetic, and
molecular genetic techniques.
Use 95% alcohol as fixative
DIGITAL PATHOLOGY & TELEPATHOLOGY
Images can be transmitted electronically to any
part of the globe.
This can be done at various levels, from the e-
mail attachment of a few static photographs to
sophisticated systems that duplicate almost to
perfection the examination of slides under the
microscope and are, therefore, accurately
referred to as virtual microscopy.
LEGAL ASPECT OF SURGICAL PATHOLOGY
Most common reasons for surgical pathologists
being brought to trial:
A mistaken diagnosis based on
misinterpretation of a slide.
An important lesion or feature present in the
specimen was missed
The pathologic diagnosis failed to give the
clinician a clear idea about the nature or
extent of the lesion or adequacy of the
sample because of poor wording or
omissions in the report
E.g. Invasion of lymphatics to determine
metastasis was not reported
Two essential components of these claims are:
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 That the alleged error or omission resulted in
physical, emotional, and/or financial damage
to the patient
That such an error or omission was below the
standards for the practice of pathology in that
particular community at that particular time.
BASIC ELEMENTS OF MALPRACTICE:
DUTY: recognition of an obligation of a physician
to treat the patient. (Physicians responsibility)
BREACH: Neglect to treat within the standard of
care. (Responsibility was not done)
PROXIMAL CAUSE: Breach causes injury in a
fairly direct manner. (neglect directly affects the
patient)
DAMAGE: Injury resulted (negative effect)
GROSS TECHNIQUES IN SURGICAL PATHOLOGY
Description of the gross specimen to include:
Measurement
Weight
Color
Consistency
Surface texture
Cut surface texture
Remember!
Once a lesion is seen intraoperatively, the surgeon must
be able to describe the lesion according to its:
5 MedTechs Before Christ
- Site location (e.g. upper outer quadrant of the
breast)
- Size in cm
- Shape standard shapes (e.g. oval, irregular)
- Surface smooth, nodular, granular, borders
- Symmetry
- Mobility
- Tenderness
- Ballotment how it feels between the thumb and
the index finger
- Consistency
FIXATION
10% buffered formalin
Relatively inexpensive
Tissue can remain in it for prolonged periods
without deterioration
Compatible with most stains including
histochemical stains
As long as the tissue is placed in fixative
shortly (<30 min) after surgical removal,
and overfixation (>2448 hours) is
avoided.
Pure formalin is 40% solution of gas
formaldehyde
Thus a 10% formalin solution represents
a 4% solution of the gas, which is 1.3
molar.
Other Fixatives:
ZENKERS
o Expensive and contains Mercuric chloride
o Often used for biopsies of the kidney, bone
marrow, lymph node and testes.
o Expensive, requires careful disposal of the
mercury, and necessitates meticulous attention to
fixation times and washing procedures to remove
the precipitates of mercury.
BOUINS
o Contains picrid acid
o Recommended for testicular biopsies
o Preservation of nucleic acids is very poor.
CARNOYS
o Mixture of ethanol, chloroform and glacial acetic
acid
o Dissolves fats
o For identification of lymph nodes in radical
resection specimens
Considerations:
Volume of fixative is at least 10X that of the
tissue.
Container should have a wide mouth - tissue can
be removed easily after it has been hardened by
the fixation.
Fixative should surround the specimen on all
sides.
Large specimens that float on a fixative
should be covered by a thick layer of gauze.
In cases of large, flat, heavy specimens that
rest on the bottom of the containers, the
gauze should be placed between the
container bottom and the specimen.
Fixation is usually carried out room temp.
For large specimen, 4C

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 Speed of penetration of formalin is 1 mm/hr.
fixation time of several hours is needed for most
specimens
o
Fixation can be hastened by heat (60 C) and by
continuous agitation by heater-motor
GROSS EXAMINATION
Specimen photography either color
transparencies or digital images
Tissue contamination (floater) some cells
retained at the cutting board from the first
specimen
Suspect a floater whenever confronted with
any of these situations:
1. A fragment of tissue that looks different
from all the others by virtue of the
thickness of the section and/or staining
intensity
2. A fragment of tissue that is on a slightly
different plane from the others, especially
if superimposed on them
3. A fragment of tissue showing pathologic
changes totally different from the others,
and of a type that one would not have
expected at all under the clinical
circumstances of the case.
st
o e.g. malignant cells coming from the 1
specimen transferred to the following
specimen giving it a false positive result
Specimen radiography for bone lesions,
calcified soft tissue masses, breast biopsies and
excisions (especially if they had been studied by
mammography), cardiac valves, and lymph node
groups in which a lymphangiogram had been
performed.
It is advantageous to fix the specimen in
Carnoy solution, which somewhat clears the
specimen by the action of the chloroform at
the same time that it fixes it.
Sampling for histologic evaluation
Must not be more than 3 mm thick
Adipose tissue must be cut even thinner.
Not larger than dimensions of tissue cassette
or glass slide
Suture material, metal clips, and other foreign
bodies should be removed from the tissues
Discrete areas of calcification or ossification
should be dissected out, or else the
specimen should be decalcified.
Better preparations will be obtained in organs
covered by folded mucosa (e.g., stomach,
bowel) if the sections are taken perpendicular
rather than parallel to the mucosal folds.
Adequate sampling with proper labeling (lead
pencil in folder paper material)
Surgical Margins
A positive margin will likely lead to local
recurrence if uncorrected.
Carried out by painting those margins with
India ink or a similar pigment before
sectioning.
The smoother the specimen contours and the
harder the consistency, the easier to identify
the true surgical margins.

Lymph node dissection - one of the most

important components in the gross evaluation of
a radical operation for cancer.
Consists of dissecting the node-containing fat
from the organ in the fresh state, using
forceps and sharp scissors.
It is too easy to crush them with the forceps
and scissors, especially if they are dissected
before fixation.

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 SPECIAL TECHNIQUES IN SURGICAL PATHOLOGY violetiodine complex (gram positive) and
The gold standard of Surgical Pathology is still with those that are decolorized by alcohol or
the use of the H & E stain on a tissue cut 5 micrometer by acetone treatment and counterstained by
a microtome. either safranin or fuchsin.
Acid Fast - depends on the high lipid content
H & E Technique - hematoxylin staining of nuclei is (mycolic acids and long-chain fatty acids) in
followed by counterstaining of cytoplasms and various the cell walls of mycobacteria, which confer
extracellular materials by eosin. to the cell the ability to complex basic dyes
- In order to function as a nuclear stain, (such as carbolfuchsin) and to retain them
hematoxylin needs to be oxidized (ripened) to following strong decoloration with acid
the purple dye hematein and provided with a net alcohol.
positive charge by combining it (chelating) with a Argentaffin and Argyrophilic stains
metallic salt (mordant) Neuroendocrine cells and their tumors, also
- Eosin is an anionic xanthene dye that combines for reticulin, melanin and calcium
electrostatically with various cytoplasmic Argentaffin reaction depends on the presence
components and with tissue such as collagen or in the tissue of a substance, often of the
muscle, the latter in an amphoteric manner. phenolic group (such as catecholamines or
- It is relatively quick, inexpensive, suitable for indolamines), that reduces silver (and other
most situations, and comparatively easy to metallic) salts
master. In the argyrophilic reaction, an extraneous
- It allows an accurate microscopic diagnosis of the reducing agent such as hydroquinone or
large majority of specimens sent to the formalin is added
laboratory. Amyloid stains
Congo red
Special Techniques for difficult slides Regarded as the most reliable and practical
Special Stains technique to detect amyloid.
Enzyme Histochemistry Reticulin Stains
Tissue Culture Reticular fibers and basement membranes
Quantitative Methods (Histometry) Reticular fibers consist of very thin fibers of
X-Ray Microanalysis mainly type III collagen, which are
Electron Microscopy widespread in connective tissue throughout
Immunohistochemistry the body.
Flow Cytometry Basement membranes are largely composed
Cytogenetics of type IV collagen and laminin.
Molecular Pathology Main applications of silver-based reticulin
stains:
SPECIAL STAINS (1) Epithelial from nonepithelial neoplasms;
PAS (Periodic Acid Schiff) Stain (2) Various mesenchymal neoplasms from
Demonstrate glycogen and neutral each other; and
mucosubstances, outlines basement (3) In situ from invasive carcinoma.
membranes and makes evident most fungi Trichrome Stains
and parasites. Evaluation of the type and amount of
Also useful for the demonstration of the extracellular material
intracytoplasmic crystals in alveolar soft part Determine parasitism
sarcoma. The three tissue structures demonstrated by
Stains for Microorganisms the three component dyes are nuclei,
Gram Stain - allows the separation of cytoplasm, and extracellular collagen
bacteria into those that retain the crystal

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 PTAH (Phosphotungstic acid-hematoxylin) stain Myelin stains
To demonstrate intracytoplasmic filaments, Luxol fast blue for the demonstration of
e.g. muscle and glial cells myelin.
Stains for hemosiderin (Perls), melanin (Fontana- Formaldehyde-induced Fluorescence - sensitive
Masson) and calcium (von Kossa) for the demonstration of catecholamines and
Perls - hydrochloric acid splits off the protein indolamines
bound to the iron, allowing the potassium
ferrocyanide to combine specifically with the ENZYME HISTOCHEMISTRY
ferric iron to form ferric ferrocyanide Fell in general disuse because of:
(Prussian blue). Complex technique
Fontana-Masson - ammoniacal silver solution Need for fresh material
is used without a reducing bath. Only Relative non specific
substances capable of reducing directly silver At present time, enzyme histochemical methods
salts (i.e., argentaffin) such as melanin are use are:
demonstrated. Skeletal muscle-related enzymes (for
von Kossa silver is substituted for calcium myopathies)
in calcium salts; this silver salt is then Acetylcholinesterase (for Hirschprungs
reduced to black metallic silver by the use of disease)
light or a photographic developer. Choloroacetate esterase (for myeloid
Stains for neutral lipids leukemia)
Oil red O is the one most commonly
employed. ELECTRON MICROSCOPY
Inconsequential distinction between fibroma Main applications
and thecoma in the ovary, support for the Renal pathology
diagnosis of renal cell carcinoma and Tumor pathology
sebaceous gland tumors of skin, and Limitations
identification of lipid-rich carcinoma in various Sampling - only a small proportion of the
organs. neoplasm can be studied
Mucin stains Paucity of truly specific ultrastructural
Used to classify gastric incomplete features - the number of organelles or other
metaplasia into subtypes (sialomucin- and structures that are exclusive of a cell or tissue
sulfomucin-containing) having supposedly type is very small
different malignant potentials Possible misinterpretation of entrapped non-
Hale colloidal iron stain has become the neoplastic elements as belonging to the
standard for the identification of renal tumor due to the difficulties in evaluating
chromophobe carcinoma spatial relationships in a small tissue sample.
Giemsa stains Diagnostic potential in:
For the demonstration of various Identification of a tumor as neuroendocrine
hematolymphoid elements (including mast through the detection of dense-core granules
cells) and microorganisms. Assessment of the nature of tumor cells with
Elastic fibers granular cytoplasm (oncocyte, granular cells
Weigert-type techniques are reasonably etc)
specific for elastin and are regarded by many ID of epithelial differentiation
as the method of choice for the ID of tumors as melanocytic through
demonstration of these extracellular fibers. detection of melanosomes
The Verhoeffvan Gieson (VVG) stain is ID of Langerhans histiocytosis through the
more popular because it is quick and outlines detection of Birbecks granules
the elastic fibers with a strong black color.

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 ID of tumor as composed of steroid-
producing cells from adrenal cortex and
gonads through the detection of abundant
smooth ER & mitochondria with
tubulovesicular cristae
ID of endothelial cells by detection of Weibel-
Palade bodies
ID of skeletal and smooth muscles by
detection of cytoplasmic filaments
ID of Schwann cells by mesoaxons
ID of alveolar soft tissue sarcoma by
detection of membrane bound crystals
ID of smooth muscles, neural or other types
of differentiation in tumors of GIST family.
Main situations where EM is used:
Differential diagnosis of carcinoma,
melanoma and sarcoma.
Differential diagnosis between
adenocarcinoma and mesothelioma
Differential diagnosis of anterior mediastinal
tumors among thymoma, thymic carcinoid,
malignant lymphoma and seminoma.
Differential diagnosis of small round cell
tumors of infancy.
Differential diagnosis of spindle cell tumors of
soft tissues.
Differential diagnosis between endocrine and
non-endocrine tumors.
IMMUNOHISTOCHEMISTRY
Application of immunologic principles and
techniques to demonstrate molecules in cells and
tissues.
Remarkable sensitivity and specificity
Applicability to routine processed material
Feasibility of an accurate correlation with the
traditional morphologic parameters
Determine specific or identifying features of the
tumors
Actin - marker for the identification of smooth muscle cells
and myofibroblasts, and for the evaluation of the
participation of myoepithelial cells in lesions of
breast, salivary glands, and sweat glands.
Epithelial Membrane Antigen (EMA) epithelial cells
- Excellent marker for most normal and neoplastic
epithelia but is not restricted to them
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- Also expressed by mesotheliomas, meningiomas,
a variety of mesenchymal neoplasms, and even
some malignant lymphomas.
- Also been found to be a marker of normal and
neoplastic perineurial cells.
Glial Fibrillary Acidic protein nerve cell
- Present in normal, reactive, and neoplastic
astrocytes; developing, reactive, and neoplastic
ependymal cells; and developing and neoplastic
oligodendrocytes
- Also been documented in peripheral nerve
sheath tumors and in mixed tumors of salivary
glands and sweat glands.
HCG gestational Trophoblastic Disease
HER2/neu (ERBB2) determines
behavior/aggressiveness of breast cancer
- When overexpressed, it acts as an oncogene.
This overexpression can be seen in breast
carcinoma (22%), lung adenocarcinoma (28%),
colorectal carcinoma (17%), lung squamous cell
carcinoma (11%), and gastric adenocarcinoma
(11%).
- Predict response to Herceptin (trastuzumab) in
carcinoma of breast and other organs
HMB-45 melanin for melanoma
- Now known to be expressed by other neural
crest-derived tumors, angiomyolipomas of the
kidney and other sites, other components of the
tuberous sclerosis complex (PEComas), and
occasional carcinomas as well as several other
neoplasms.
Keratins squamous cells
- Excellent marker for epithelial differentiation
regardless of whether the tumor is of
endodermal, neuroectodermal, mesenchymal, or
germ cell derivation
- Particularly common and prominent in synovial
sarcoma, epithelioid sarcoma, and uterine
smooth muscle tumors
Neuron Specific Enolase - found in the majority of
neuroectodermal and neuroendocrine
neoplasms, including carcinoid tumors and
malignant melanoma.
PSA prostate cancer
- Method of choice for the identification of prostatic
adenocarcinoma.
S-100 malignant melanoma
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 - Evaluation of peripheral nerve sheath and translocations has been particularly
melanocytic tumors. successful in the fields of leukemias and
lymphomas, germ cell tumors, pediatric
FLOW CYTOMETRY tumors, and mesenchymal neoplasms
Measurement of various parameters while a Contributions of CytoGen to Pathology:
suspension of cells flows through a beam of light (1) Defining subsets within putatively
past stationary detectors. histologically homogeneous tumor types
Light scattered at various angles by the cells is (2) Suggesting connections between
registered by detectors and converted to histologically diverse tumors
electronic signals and analyzed by computers (3) Highlighting specific changes in
Cellular features evaluated include cell size, histological subtypes
cytoplasmic granularity, cell viability, cell cycle (4) Suggesting the site of the primary when a
time, DNA content, surface marker phenotype specific cytogenetic change is found in a
and enzyme content. metastasis
Usually used in hematopathology (5) Providing clues to tumor classification,
causation, and presence of cancer-
related genes.
MOLECULAR PATHOLOGY
Based on the application of recombinant DNA
technology.
Can be performed in tissues handled as part
of routine work, including formalin-fixed,
paraffin-embedded material.
Snap frozen material is superior in regard to
the yield and quality of extracted DNA and
particularly RNA
Stained cells enter the flow chamber where they
pass into the center of a stream of sheath fluid in single
file. They are then struck by a focused laser beam and
emit scattered and fluorescent light, which is separated
according to wavelength by appropriate mirrors and
filters. An obscuration bar protects the forward angle light
scatter (FALS) detector from exposure to the direct laser
beam. Only two fluorescence detectors are shown, for
sake of simplicity, but a typical instrument has three or
four such detectors, one of which can be used to
measure laser light scattered perpendicular to the laser
beam by the cells. Signals from the detectors pass to
amplifying processors and then to the integral (on-board)
computer, which digitizes the signals, stores them, and
displays them.

NEWER TRENDS
CYTOGENETICS
Karyotypic analysis of human tumors
Detection of nonrandom or specific
chromosome abnormalities such as
deletions, amplifications, inversions, and

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