12/6/2011

COURSE : CLINICAL PATHOLOGY

TOPIC : EXFOLIATIVE CYTOLOGY
SUBMITTED TO,
DR SUGUNA RAO
PROFESSOR
PRESENTED BY,
PRIYA S
MVHK
MVHK-- 1039

The accuracy of the cytologic

examination depends on the
Quality of collection
Preparation
Staining and
Interpretation of the material.
Inadequacy in any of these steps will
adversely affect the quality of
diagnostic cytology.

Making quality smears of

impression smears

Exfoliative cytology :
A branch of General Cytology which deals
with the microscopic study of cells that
have been desquamated
from the
epithelial
surfaces
and
mucous
membranes..
membranes
George
N
Papanicolou
introduced
cytology as a tool to detect cancer and
pre-cancer in 1928.

Collection Methods
1. Imprints
Touch imprints may be made directly from
crusted and ulcerative skin lesions or from
impressions of deeper surgical biopsies
gently rolled onto a glass slide prior to
placement in formalin.
Dry scabs/crusts should be removed
manually prior to impression smears being
made, as cells in these scabs/crusts will
generally reveal poor cellular morphologic
preservation
and
poor
staining
characteristics.

2. Scrapings

The advantages of diagnostic cytology

are
Non-invasive
Simple procedure
Helps in faster reporting
Relatively inexpensive
Has high population acceptance and
facilitates cancer screening in the
field.

The tissue should be blotted dry (using paper towel) to remove

surface fluid or blood as these may impair adhesion of cells to
the slide and dilute the cytological material.
The biopsy sample or lesion being examined is firmly pressed
several times onto a clean glass slide. Do not rub the tissue on
the slide, as this will result in distorted cellular morphology by
causing cell rupture and nuclear stranding.
Imprints from ulcerative lesions often only yield superficial
inflammation and infection and any underlying/primary
neoplastic process may be missed.
Advantages :
Good technique for investigating the presence of inflammation
with or without infectious agent involvement
Also useful in assessing the presence of superficial neoplasia
(squamous cell carcinoma).

Making quality smears of tissue

scrapings

The back (blunt edge) of a scalpel blade or

edge of a glass slide is used to gently
scrape across the lesion or tissue biopsy
until a small amount of material is
collected. This material is then gently
spread across a slide.
This method may be used where imprinting
is likely to yield too few cells for complete
assessment
(e.g.,
conjunctiva,
mesenchymal neoplasia).

12/6/2011

3. Swabs
This technique is useful for the sampling of
fistulous tracts, ear canals, exudates and
for vaginal cytology.

Making quality smears with cottontipped swab

4. Fine Needle Biopsy / Fine Needle

Aspirate Biopsy (FNAB)
The best and most commonly used
method for sampling proliferative lesions
and masses.

Unless the location is very moist, lightly

moistening the swab with 1 or 2 drops of
sterile saline prior to collection is advised,
as this will minimise cell damage.

Use of 22-25 gauge needle and a 2-5 ml

syringe recommend and as a general rule,
the softer the tissue, the smaller the
needle and syringe required to obtain an
adequate sample.

Smears are prepared by gently rolling the

swab over a glass slide. Do not smear the
swab on the slide in a side-to-side motion
as this causes cell rupture and poor cell
preservation.

Aspiration procedure:
Once the mass is stabilised between the operators fingers,
the fine gauge needle is inserted into the mass.
When the needle is seated comfortably in the mass,
negative pressure is applied to the plunger/syringe.
Try to avoid redirecting the needle or moving it back and
forth within the mass whilst vacuum (negative pressure) is
applied, as this generally results in increased blood
contamination of samples.
This procedure should be repeated at least 3 4 times at
different angles within the lesion to obtain a representative
cell population from the lesion.
Smaller syringes attached to the needle offer the operator
better control during the aspiration process, particularly
when aspirating smaller lesions.

A minimal amount of material within the hub of

the needle is adequate and generally, this is
sufficient for cytological interpretation.

Non-aspiration procedure:

Attempted further aspiration often leads to

unwanted blood contamination. If blood is
encountered during aspiration attempts, then the
exercise should be ceased and repeated a little
further away from the initial puncture site.

This technique also utilises a fine gauge needle

(22 25 g) but syringe is not attached to the
needle.
Applied for sampling highly vascular masses, as
blood contamination is often reduced(skin
masses).

Negative pressure should be released before the

needle is removed from the mass and skin.

The needle is briskly redirected within the mass

at several different angles.

Once the needle is removed from the syringe, air

is drawn into the syringe and the needle is firmly
re-attached to the syringe.

Once the needle is removed from the lesion, it is

attached to an air-filled syringe and the material
within the needle is gently expelled onto clean
glass slide/s for smear preparation.

The material within the hub of the needle is then

expelled onto a couple of slides for smear
preparation.

PRECAUTIONS WHILE COLLECTION

FOR CYTOLOGY SPECIMENS:

Preparation of Slides
The aim of slide preparation for
cytological evaluation is to achieve a
monolayer of well-preserved cells.

1. Avoid blood contamination. Possibly use the non-aspiration needle

biopsy technique for soft, highly vascular and small lesions.
2. Do not prolong the period of aspiration (should take less than 30
seconds) and make smears immediately after collection to optimise cell
preservation.

Redirect
needle
3-4
times when sealed in
the mass and if using
vaccuum
dont
pull
back on plunger whilst
redirecting needle

Pin cushion non

aspiration

3. Attempt 2-3 separate collections (if the lesion/mass is large enough).

Several methods:

4. Make 2 slides from each collection.

5. If cell yield appears poor via FNAB, use a larger needle and syringe
and/or increase the amount of negative-pressure within the syringe.
6. Material within the hub of the needle is usually sufficient and further
sampling often results in unwanted blood contamination.

1.
2.
3.

Squash Preparation
Needle/Starfish Preparation
Blood
Smear
Technique

7. when blood appear in the syringe during FNAB, stop the procedure
immediately and start again with a new needle and syringe.

12/6/2011

1. Squash Preparation (A misnomer?)

This is the preferred method for preparing slides from needle
biopsies, FNAB and scrapings.
Material collected by fine needle biopsy or scraping is placed
towards one end of a glass slide. And a second slide is aligned
perpendicular to the first and is allowed to rest on the slide
containing the expelled material.
The second slide is then gently and smoothly drawn over the
length of the first slide whilst concurrently rotating them from
a perpendicular to a parallel position. This results in the
simultaneous preparation of two smears.
Avoid physically squashing the material, which causes
excessive pressure, leading to cell rupture and a nondiagnostic preparation.

PRECAUTIONS FOR SLIDE PREPARATION

Ensure clean glass slides are used and minimise handling of
glass slides to reduce collection artefacts.
Prepare two slides from each collection.
Be gentle when preparing squash preparations.
Rapidly drying the slides (hairdrier or by waving in the air)
after preparation, reduces cell crenation & slow drying
artefact, resulting in superior cell preservation.
The presence of a large amount of blood or the preparation
of thick smears also severely hinders cytological cell
evaluation.
Avoid placing material near the edges of the slides,
particularly towards the shorter edges, as these regions may
be difficult to assess microscopically.
Label the slides and not the slide carriers.

PRECAUTIONS FOR SUBMISSION OF

CYTOLOGY SPECIMENS:

2. Needle/Starfish Preparation
Material collected by fine needle biopsy is
placed in the centre of a glass slide and the
needle is used to drag/tease the material
outwards in multiple directions - to
produce a star/starfish shaped smear with
multiple projections.
Many areas of the smear will be too thick
for evaluation, however, there are usually
multiple cell monolayer regions present on
the smear that should be acceptable for
cytological assessment.

Collection and Preparation of Slides from

Fluid Samples
Fluid samples are obtained from sampling body cavity effusions,
cysts, joints, cerebrospinal fluid, urine and when performing
various types of washes (e.g., bronchoalveolar lavage,
transtracheal wash).
Fluid should be collected into EDTA-containing tubes and
sterile/plain tubes. Smears should also be made at the time of
sampling.
Making smears at the time of sampling also helps better
determine the relevance/significance of certain cytological
features such as erythrophagocytosis and the presence of intracellular bacteria (phagocytosis of both red cells and bacteria by
leukocytes may occur post collection during transit).
EDTA is bacteriostatic and culturing from EDTA-containing fluid
is generally not advised.

3. Air-dried smears made at the time of sampling do not require further

fixation and will survive transit to the laboratory as is.
4. Submit slides in rigid plastic slide-holders and not in cardboard holders
or by any other method (e.g., wrapped in surgical swabs or tissues).
5. Submitting unstained smears is preferred, however, it is often
advantageous to stain one smear in-house to assess the adequacy of
cellularity and smear preparation prior to submission to the
laboratory.
6. Do not submit unstained smears with formalin-containing specimens.
Formalin vapour will fix unstained material and will severely
adversely affect subsequent staining intensity and quality, and will
often render the smears non-diagnostic.

The material is expressed towards one end of a glass slide

and the short edge of the spreader slide is placed in front of
the sample.
The spreader slide is tilted to an angle of approximately 45
degrees, pulled backwards into the material and once the
material has dispersed along the width of the spreader slide,
the spreader slide is smoothly, steadily and rapidly slid
forward. The smear ends with a feathered edge of material.
As a general rule, the more material placed on the specimen
slide, the slower the spreader slide is slid forward and the
more acute the angle between the spreader and specimen
slide, the longer the smear will be.

If there are any floccules of particulate matter grossly visible in

the fluid at the time of collection, then these should be
included in the smears as well.
Aliquots of clear or slightly turbid fluids should be concentrated
via centrifugation to increase the cellularity of prepared
smears.
Following centrifugation, the majority of the supernatant is
decanted, the cellular material is then resuspended in the
minimal remaining supernatant, and smears are made via
either the blood smear techniques or line smear technique.
The line smear technique is similar to the blood smear
technique; however, the spreader slide is abruptly stopped and
lifted off the specimen slide prior to creating a feathered edge,
resulting in a higher concentration of cells present in the
terminal line, than within the remainder of the smear.

Concurrent submission of a plain fluid sample collected into a

sterile container, allows for culture to then be performed if initial
cytology indicates that this may be warranted.

Staining of Cytology Smears

Several different types of stains are used for
cytology: Romanowsky type stains (Wrights,
Giemsa), Supravital stains (toluidine blue, New
Methylene blue) and Papanicolaou stains.

Romanowsky stains are inexpensive, easy to use

and they are readily available to veterinary
practitioners. They provide good nuclear detail,
excellent
cytoplasmic
detail
and
infectious
organisms are readily visualised.

Papanicolaou stains provide excellent nuclear detail

and adequate cytoplasmic detail, however, they are
time consuming and impractical for in-clinic usage.

In clinical practice, the most cost effective,

quickest and easiest stain to use is the Giemsa
stain.

Supravital stains provide excellent nuclear detail but

poor cytoplasmic detail and are typically reserved for
evaluation of reticulocyte identification (peripheral
blood smear) or for evaluating the presence of
poorly granulated mast cells.

As a general rule, the thinner the material on the

smear, the less time needed for staining, and the
thicker the material, the more time required for
staining.

1. Submit 2-4 well-prepared smears from each location sampled.

2. Label all smears appropriately.

3. Blood Smear Technique

Aspirated material may contain enough blood and/or liquid to
allow smearing of the material in a similar fashion to that
used to make a peripheral blood smear.

7. When submitting fluid samples, submission of in-house made smears,

EDTA-containing fluid and fluid in a plain/sterile container is
recommended.

12/6/2011

PRECAUTIONS WHILE STAINING CYTOLOGY

SPECIMENS:
1. Sealing staining solution containers properly (i.e., can use
cling-film between the lid and container) will minimise the loss
of stain fluid due to vaporisation.
2. Make sure that the smears are completely dry before staining.
3. Take care not to over-stain smears.

TIPS FOR DETERMINING THAT A SMEAR IS ADEQUATE FOR

CYTOLOGICAL EXAMINATION AND INTERPRETATION:
6. If bacteria and/or fungi are regularly noted within

examined smears (particularly if sampled sites are

not superficially ulcerated lesions), then the
staining solutions are likely contaminated and need
to be replaced. This can be confirmed by staining
an unused, clean glass slide and then examining
this slide for the presence of organisms.

4. Under-stained smears can be re-stained for further periods to

increase staining intensity.
5. Stains will lose potency over time and need to be replaced.
6. Stain precipitate typically present as extracellular blue granular
material that is often clumped. The stains should be mixed
thoroughly or replaced.

Vaginal cytology
Vaginal cytology is a simple technique that can
be used to characterize stages of the
reproductive cycle of the bitch or to evaluate
certain diseases of the genital tract
tract..

Vaginal cytology usually is used :

to determine the stage of the reproductive cycle
for artificial insemination
insemination..
for determination of the whelping date
diagnosis of inflammation of the vagina and
identification of some types of neoplasia involving
the vagina and urethral orifice
orifice..

7. Evaluate the smears for the thickness of material

and the staining intensity and quality, and adjust
future staining times appropriately.

Basal Cells
Cells:: Basal cells are the youngest cells of the vaginal
epithelium and serve as precursors of the other epithelial cell
types
types.. They are located along the basement membrane and are
rarely seen in exfoliative cytology
cytology.. These cells have a high
nuclear to cytoplasmic (N/C) ratio, a round nucleus, and a
small amount of basophilic cytoplasm
cytoplasm..
Parabasal cells:
cells: These cells are the smallest cells of vaginal
epithelial cells seen in cytologic preparations
preparations.. Parabasal cells
also have a high N/C ratio, round nucleus, and basophilic
cytoplasm.. They are uniform in size and shape
cytoplasm
shape..
Parabasal cells also may contain cytoplasmic vacuoles and are
known as foam cells.
cells. The function of the vacuoles in these cells
is unknown
unknown.. Large numbers of foam cells may be found in
prepubertal samples.

2. Scan the smear at low power (4x or 10x objective) to assess

overall cellularity and to find areas within the smear that
contain a monolayer of well-preserved and adequately stained
cells.
3. Examine smear using high dry technique (40x objective) by
placing a coverslip directly on the dry stained smear. Oil is
not required.
4. Use high power (40x objective) to evaluate individual cells
within the monolayer.
5. Immersion oil is usually only required to investigate the
presence and morphology of infectious agents such as
bacteria.

Collection and Preparation of Specimens

A saline moistened cotton swab is inserted through the vulvar
lips into the vagina
vagina.. The swab is angled craniodorsally to avoid
the clitoral fossa
fossa.. A clean otoscope may be used to guide the
swab and to provide a light source for visual guidance
guidance..
Once the swab is located cranial to the urethral orifice, the
swab is rotated slightly to exfoliate and collect the cells.
cells.

A drop of fluid is placed on a glass slide and a

bloodblood-type smear is made, air dried, and stained
stained..
The medicine dropper technique is more gentle
but the saline will dilute the cell count
count..

After the swab is removed from the vagina, the cells are
transferred to a clean glass slide by rolling the swab along the
surface of the slide.
slide.

Various stains have been used for vaginal

cytology including Romanowsky stains (Wright,
Giemsa
Giemsa,, Leishman),
Leishman), new methylene blue,
toluidine blue, trichome stains (Papanicolaou,
Papanicolaou,
Shorr
Shorr,, Sano) and hematoxylin and eosin (H&E).
(H&E).

An alternative method to collect cytologic material from the

vagina is to use a plastic medicine dropper and saline solution.
solution.
The dropper is passed into the vagina and the bulb is squeezed
several times
times.. Exfoliated cells are collected by saline lavag
lavag..

The stain should be economical, easily stored,

and produce consistent staining that is adequate
for cytologic needs
needs..

Cytologic Features of Vaginal Epithelial

Cells
The vaginal epithelium is influenced by hormonal changes
during the estrous cycle, allowing cytologic monitoring of the
various reproductive stages
stages..

1. Allow the smear to dry completely prior to examination. This

may be enhanced by using a hair drier set on the low heat
level.

Cytologic Staging of the Canine

Reproductive Cycle
Intermediate cells:
cells: These cells vary in size but are

usually two times the size of parabasal cells.

cells. Their N/C ratio
is decreased and they have large amounts of blue
blue--green,
keratinized cytoplasm.
cytoplasm.
Their borders are round to irregular and folded
folded..
Intermediate cells also may be subclassified as superficial
intermediate or transitional intermediate cells according to
their stage in the reproductive cycle
cycle..

Superficial cells: Superficial cells are the oldest

vaginal epithelial cells.
cells. They either have small, round,
pyknotic nuclei or lack a nucleus if they are cornified
cornified.. Their
cytoplasm is abundant, blue
blue--green, and keratinized.
keratinized. Cell
margins are angular with folded edges
edges..

Each stage of the canine reproductive cycle has a

distinct, predominant cell population.
population. that is
easily identified cytologically
cytologically..
Proestrus: Predominant cell populations in early
Proestrus:
to
mid
proestrus
include
nondegenerate
neutrophils and a mixture of parabasal,
parabasal,
intermediate, and superficial epithelial cells.
cells.
In late proestrus,
proestrus, the number of neutrophils
declines
and
superficial
cells
begin
to
predominate

12/6/2011

Early proestrus with a mixture of

nondegenerate
neutrophils;
parabasal,
intermediate,
and
superficial
epithelial
cells,
erythrocytes, and mucus (WrightLeishman stain).

Late proestrus with erythrocytes, scattered

superficial cells, and absence of neutrophils
(Wright-Leishman stain).

Diestrus:: Cell populations change abruptly in

Diestrus
diestrus.
diestrus.
Superficial cells decrease by 20
20%
% and smaller
intermediate cells increase in number.
number.
Neutrophils often reappear and may contain
phagocytosed erythrocytes and bacteria.
bacteria.
Cytologically,, this stage of the reproductive cycle
Cytologically
can look very similar to early proestrus.
proestrus. Therefore,
serial cytologic sampling is required to make the
distinction..
distinction

Estrus: In estrus,
Estrus:
estrus, the cell population
consists of ~ 90
90%
% superficial cells
and < 5% parabasal or intermediate
cells.
cells.

folded cell margins (Wright(Wright-Leishman stain)

stain)..

Less mucus is present

present.. Normal
bacterial flora usually is present and
organisms often are attached to the
superficial cells.
cells.

Diestrus characterized by a decrease in superficial cells,

increase in smaller intermediate cells, and presence of
neutrophils.. (Wrightneutrophils
(Wright-Leishman stain).
stain).

Vaginitis
Cytologic changes in vaginitis include the
presence of many degenerate or nondegenerate
neutrophils..
neutrophils
Mucus, lymphocytes, and macrophages also may
be present and may indicate chronicity.
chronicity.
If vaginitis is due to an infectious agent,
phagocytosed bacteria or intracnuclear inclusions
also may be observed.
observed.

Anestrus::
Anestrus

Parabasal and intermediate cells

predominate during anestrus.
anestrus.
Superficial cells are not present.
present.
Erythrocytes and neutrophils may be present in low
numbers or are absent
absent..

Vaginitis demonstrating the presence of many

nondegenerate to degenerate neutrophils and
scattered epithelial cells (Wright
(Wright--Leishman stain)
stain)..

Estrus characterized by superficial cells that are

keratinized, largely anucleate,
anucleate, and have angular,

An abundance of neutrophils may be present in

diestrus.. Therefore, this normal stage of
diestrus
reproduction
must
be
distinguished
from
inflammation..
inflammation

Vaginal Neoplasia
Vaginal neoplasia is the second most common form of
reproductive neoplasia in the bitch, following mammary
neoplasia..
neoplasia
Vaginal neoplasia includes transitional cell carcinoma (TCC) of
the urethra, transmissible venereal tumor (TVT), squamous cell
carcinoma (SCC), leiomyoma,
leiomyoma, and fibropapilloma.
fibropapilloma.
Vaginal tumors are usually found in the older bitch except for
TVT which is more common in young dogs.
dogs. The mean age of
occurrence of vaginal neoplasia in the bitch is ~10 years
years..
Bitches with vaginal neoplasia typically present because of a
mass protruding from the vulva, vaginal discharge and dysuria
dysuria,,
stranguria,, tenesmus,
stranguria
tenesmus, urinary incontinence, or perineal swelling
due to mechanical interference by a tumor
tumor..

Vaginal cytology is most helpful in diagnosing TVT,

TCC, and SCC.

Transmissible venereal tumor

TVT is transmitted sexually, by mucous membrane
contact (i.e,. sniffing and licking), or via bite wounds
and tends to be common in areas where dogs roam
freely
freely..
These neoplasms are either benign or malignant in
behavior..
behavior
Metastasis is rare but may involve regional lymph
nodes, skin, and subcutanteous tissues
tissues..
TVT cells have a round nucleus, single nucleolus,
and a thin rim of light blue, vacuolated cytoplasm.
cytoplasm.
TVTs may resolve spontaneously without treatment
treatment..
Treatment with vincristine or irradiation is very
effective..
effective

12/6/2011

TVT cells with a round nucleus, single nucleolus,

and a thin rim of light blue, vacuolated cytoplasm
(Wright--Leishman stain)
(Wright
stain)..

Transitional cell carcinoma.

carcinoma. The epithelial cells exhibit
anisocytosis,, anisokaryosis
anisocytosis
anisokaryosis,, basophilic cytoplasm and have

TCC & SCC

Transitional cell carcinomas and SCC are common
in the distal urethra of the bitch and may involve
the vagina or vestibule or both
both..

an increased N/C ratio (Wright(Wright-Leishman stain).

stain).

Vaginoscopy and surgical biopsy are best for a

definitive diagnosis.
diagnosis. Occasionally, fine
fine--needle
aspirates may provide a definitive diagnosis of TCC
or SCC.
SCC.
Cells in TCC exhibit anisocytosis
anisocytosis,, anisokaryosis,
anisokaryosis,
and have an increased N/C ratio
ratio..
Neoplastic squamous cells are characterized by
anisocytosis,, anisokaryosis,
anisocytosis
anisokaryosis, and waxy
blue
cytoplasm, occasionally with a perinuclear halo
halo..
Cytologic evaluation of urine sediment is helpful
but is only diagnostic in ~ 40
40%
% of cases
cases..

Squamous cell carcinoma

carcinoma.. The neoplastic squamous cells
exhibit anisocytosis,
anisocytosis, anisokaryosis,
anisokaryosis, and waxy blue
cytoplasm.. Neutrophils and erythrocytes also are present
cytoplasm
(Wright
(Wright--Leishman stain)
stain)..

Cervical smear
Cancer of the uterine cervix
commonest cancer in the FGT.

Preparation of Smear
is

the

Sampling Devices:
Ayres spatula
The Cervex brush device is a flexible
plastic brush, which follows the shape of
the cervix.

Cytologic examination of the urine

sediment
Different types of epithelial cells can be observed in
urine
urine.. Some of these are readily identifiable,
however it is difficult to distinguish small transitional
epithelial cells from WBC and renal tubular epithelial
cells from transitional epithelial cells.
cells.
Epithelial cells are semi
semi--quantified in urine as
as:: none,
seen - few, moderate, many
many..
Neoplastic cells of renal, urinary or reproductive
origin can exfoliate in the urine and a urinalysis is
definitely indicated if a tumor in one of these sites is
suspected in the animal
animal..

Transitional epithelial cells

Squamous epithelial cells

The urinary tract from the pelvis down the

ureters to the bladder and the proximal urethra is
lined by transitional epithelial cells.
cells.

These can be keratinized or non

non--keratinized.
keratinized.

These cells vary in size and shape depending on

the location from which they originate, e.g. those
from the renal pelvis are more caudate whereas
those from the bladder are more round to
polygonal and vary in size.
size.

Non
Non--keratinized squamous epithelial cells originate
from the distal urethra, prepuce and/or vagina
vagina..
They are larger than transitional cells and have
small central nuclei.
nuclei. They can be round or have
one or more flat border
border..

These cells naturally slough into the urine in quite

low numbers, so none to a few transitional
epithelial cells are seen in the urine from healthy
animals.. This depends on the method of urine
animals
collection, since these cells will be sloughed
(traumatically) when the bladder is catheterized.
catheterized.

Keratinized squamous epithelial cells are from the

skin or vulva and are large cells with angular
borders
borders.. They may or may not have nuclei.
nuclei.

12/6/2011

Renal tubular epithelial cells

Squamous cells are frequently seen as contaminants
in urine samples.
samples.
Urine collected by cystocentesis donot contain any
squamous epithelial cells.
cells.
Although these cells are considered contaminants,
large
numbers
may
represent
abnormal
genitourinary conditions, specifically squamous
metaplasia of the prostate in the dog.
dog.
This occurs secondary to excess estrogen,
estrogen, usually
secreted by testicular tumors (particularly Sertoli
cell tumors
tumors)).

Neoplasia
Neoplastic cells, typically those of transitional
epithelial origin (transitional cell carcinoma or
TCC) may slough into the urine
urine..
The presence of these cells can be diagnostic of
urinary neoplasia,
neoplasia, however they are not always
seen in the urine in affected animals (i..
..e
e. the
lack of these cells in a urinalysis does not rule out
neoplasia)).
neoplasia
TCC are more common in dogs and frequently
originate in the trigone of the urinary bladder,
although the prostatic urethra is a common site
in male
male..

The upper panel demonstrates large neoplastic

epithelial cells in an unstained urine sediment from a
dog with a bladder mass.
mass. The lower panel
demonstrates that these cells display clear cytologic
criteria of malignancy in a Wright's stained cytology

Diagnosis
of
neoplasia
depends
upon
the
identification of cytologic criteria of malignancy in the
epithelial cells, e.g. marked variation in nuclear and
cell size, multiple nucleoli of variable size within one
nucleus,
multinucleation
with
intracellular
anisokaryosis,, macronucleoli.
anisokaryosis
macronucleoli.

These are rarely seen in the urine and are very

difficult to distinguish from transitional epithelial
cells.
cells.
If large numbers of smaller epithelial cells of
uniform appearance (size and shape) are observed
in the urine, a renal origin for these cells is
suspected..
suspected
Transitional epithelial cells tend to be more
variable in size and shape (to some extent).
extent).
Sloughing of large numbers of renal tubular
epithelial cells would indicate renal tubular injury
injury..

Transitional cell carcinoma, urine sediment, dog,

Wright
Wright--Leishman stain
stain.. Bacteria and purulent
inflammation are present in conjunction with
neoplastic epithelial cells.
cells.

TCC usually invade the bladder wall and cause

hemorrhage..
hemorrhage
They may also become secondarily inflamed from a
superimposed bacterial infection or necrosis.
necrosis. Thus,
hematuria and, to a lesser extent, pyuria,
pyuria, can be
features of a urinalysis in animals with TCC.
TCC.
Rarely, other tumors originating in the bladder or
kidney (e
(e..g. lymphoma, renal carcinoma) can
exfoliate into the urine
urine..

Transitional cell carcinoma, dog, WrightWrightLeishman stain

stain.. Neoplastic cells are
scattered singly or arranged in variablyvariablysized clusters
clusters..

Transitional
cell
carcinoma,
dog,
Wright
Wright-Leishman stain
stain.. Anisocytosis,
Anisocytosis, anisokaryosis,
anisokaryosis, and
variation are present within the neoplastic
epithelial cells.
cells.

smear of the urine.

urine.

12/6/2011

Transitional cell carcinoma, dog, WrightWrightLeishman

stain
stain..
Variation
in
the
abundance of cytoplasm among neoplastic
transitional cells.
cells.

Transitional
cell
carcinoma,
dog,
Wright
Wright-Leishman stain
stain.. Scattered necrotic cells stain
light
gray
and
lack
discernable
nuclei.
nuclei.
Degenerative neutrophils,
neutrophils, bacteria, and Bilirubin

Transitional cell carcinoma, dog, WrightWrightLeishman stain

stain.. Multinucleated neoplastic
transitional epithelial cell (right side of
image)..
image)

crystals also are present.

present.

Synovial Fluid Analysis

Concentrations of synovial fluid electrolytes and nonelectrolytes,
nonelectrolytes,
such as glucose and urea, are similar to those of blood.
blood. It has a
relatively low protein concentration
concentration.. The cells of the synovial
membrane
add hyaluronate,
hyaluronate,
a
glycosaminoglycan
that
contributes to viscosity of the fluid.
fluid.

APPEARANCE :
The appearance of synovial fluid is characterized by its color,
color,
turbidity, viscosity, quantity, and ability to clot.
clot.
Note whether the fluid was bloody initially or became so during
aspiration.. The sudden appearance of blood in the synovial fluid
aspiration
during aspiration is also a reliable indication of a traumatic tap
tap..
In the atraumatic tap yielding bloody synovial fluid, the fluid
appearance does not change during the aspiration but remains
bloody throughout
throughout..

Dark yellow or xanthochromic fluids may indicate

chronic hemorrhage and erythrocyte breakdown
and the formation of bilirubin compounds
compounds..

Lowered viscosity, regardless of the synovial fluid

appearance,
is
usually
indicative
of
an
inflammatory change
change..

Turbid samples of varied color are commonly

associated with inflammatory joint disease, which
may be septic or nonseptic.
nonseptic. Turbidity is usually due
to cells, fibrin, or other debris
debris..

Normal synovial fluid does not clot, Synovial fluid

that clots suggests the presence of synovitis and
is caused by fibrinogen and other clotting factors
in the fluid that are not present normally.
normally.

Synovial fluid viscosity is easily assessed by slowly

expelling one or two drops of fluid from the syringe
through the needle
needle.. If the fluid flows with the ease
of water the viscosity is low.
low.
Formation of a tenacious string as the drop leaves
the needle indicates normal viscosity.
viscosity.

Parameter Normal joint synovial fluid

CYTOLOGIC EXAMINATION
Cytologic examination of synovial fluid is similar
to that of peripheral blood in that total numbers
of leukocytes and erythrocytes are counted
counted..
The diluent used for blood counts cannot be used
for synovial fluid because it will cause mucin
precipitation and thus alter the count
count..
Physiologic saline used as a diluent.
diluent.
Pretreating highhigh-viscosity synovial fluids with
hyaluronidase has been recommended for the
even distribution of cells for counting and
identification..
identification

Normal cell counts of synovial fluid of the dog

range from 0 to 3000
3000/mm
/mm3
3.
Mononuclear cells and lymphocytes predominate.
predominate.
Occasionally monocyte
monocyte--macrophage type cells are
seen
seen..
Neutrophils are rarely present and when seen are
usually the result of blood contamination during
sampling.. Increased numbers of neutrophils are
sampling
indicative of disease
disease..
An occasional erythrocyte may also be present
owing to sampling contamination.
contamination.
Wright's stain and new methylene blue are
commonly employed for synovial fluid smears
smears..

Osteoarthritis joint synovial

fluid

Color

Transparent and colorless to yellow

Usually normal

Turbidity

None

None to slightly turbid

Viscosity

Very viscous
3-5cm string

Normal to mildly decreased

viscosity;
1-2cm string

Cell count

Overall low cellularity; red blood cells

absent, small number nucleated
cells
<500/uL equine<1000/uL bovine
<1500/uL (dog/cat)

Normal to slightly increased

population of mononuclear
cells
>500/uL equine>1000/uL
bovine
>1500/uL (dog/cat)

Neutrophi
ls

<6-12%

Variable, normal to increased

Mononucl
ear
cells*

90-100%

90-100%

Total
protein

1.8-4.8 g/dl (generally <2.5 g/dl)

Increased >2.5 g/dl

Other
feature

No toxic cells or microorganisms

microorganisms

12/6/2011

Respiratory cytology
Samples obtained for cytology and culture from the respiratory
tract by fine needle aspiration, washing and brushing.
Cytology of respiratory secretions is used in the diagnosis and
characterization of respiratory disease
disease..

Cytology :
Ciliated epithelial cells (tall columnar to columnar/cuboidal
columnar/cuboidal))
representing all levels of the airway (more in samples collected
by brushing).
brushing).
Macrophages: predominate in lavage fluids from healthy
Macrophages:
horses
horses..
Non
Non--degenerative polymorphonuclear neutrophils (PMNs) and
lymphocytes:: numbers generally low in health
lymphocytes
health..
Variable amounts of mucus
mucus..

Cell types description

NonNon-cellular material
Mucus :

Squamous epithelial cells:

Source is the upper respiratory tract lining, pharynx and
larynx
larynx..
Indicates No particular pathologic significance
significance..

Epithelial cells:
cells:
The airway epithelial cells from the upper trachea to the
terminal bronchioles are ciliated and become progressively less
tall towards the smaller airways
airways..
Normal epithelial cells have a basal nucleus with visible
chromatin pattern and a finely vacuolated cytoplasm.
cytoplasm.
The cilia, which are attached to a well defined terminal plate,
are fine, separate and evenly proportionate in length
length..
Sometimes difficult to see in clusters of cells.
cells.

Amount is noted
noted..
recurrent airway obstruction (RAO) - horses may show
layered mucus with casts and embedded cells.
cells.

Inflammatory cells:
cells:
In septic bronchopneumonia, many (40
40--90
90%
%) degenerate
polymorphonuclear leukocytes (PMNs, neutrophils),
neutrophils), some of which
may contain intracellular bacteria and epithelial cell necrosis
necrosis..
Similarly high proportion of PMNs, mostly non
non--degenerate, and
numerous reactive and multinucleate macrophages.
macrophages.

Lymphocytes::
Lymphocytes

Extraneous non
non--cellular material
material..
Plant or fungal decontaminant may be seen in normal or
dysphagic horses
horses..

Alveolar macrophages :
Part of the mononuclear phagocyte system, adherent to
alveolar wall.
wall. Protect respiratory membrane and have
phagocytic function
function..
Active macrophages show vacuolated or foamy
cytoplasm and increased stippling of the nucleus
nucleus.. A few
multinucleate macrophages may be seen in normal
washes
washes..

Macrophages:
Active macrophages have prominent vacuolation of the
cytoplasm and they become multinucleated with chronic
irritation.
irritation.
The nature of engulfed material provides information on the
type of activity of the macrophage.
macrophage. Can see engulfed fungal
hyphae and spores, pollen grains, bacteria, carbon particles
particles..
Hemosiderin laden macrophages support exercise induced
pulmonary hemorrhage (EIPH) - but may also occur with
severe congestion, vascular damage or other causes of
hemorrhage..
hemorrhage

Few seen in normal washes, more in allergic small airway disease

disease..

Fungal elements
elements::
Eosinophils::
Eosinophils
Eosinophils are not normally found in tracheal aspirate samples
samples..
Their presence in significant numbers are seen with helminth
infections (lungworm ) and in some cases of inflammatory airway
disease .

The level of airway involvement is estimated from

the type of epithelial cells showing degenerative
changes..
changes

Debris :

Goblet cells:
Rarely seen in normal washes.
Presence of increased numbers of active (foamy) goblet cells may
indicate irritation and excessive mucous production.
production.

Abnormal cells
Epithelial degeneration :

Free or engulfed (within macrophages) fungal hyphae and

spores
result
from
inhalation
with
environmental
contamination, as well as with pathogenic fungal infections.
infections.

Epithelial cell degenerative changes include:

include:
Clumping and stunting of cilia or Loss of cilia.
cilia.
Disruption of the cell, distortion of its normal shape
(dysplasia, metaplasia)
metaplasia).
The term "creola body" is used to describe metaplastic
bronchiolar epithelial cells, usually a response to chronic
irritation, eg allergy
allergy..
Pyknosis of the nucleus, uneven vacuolation of the
cytoplasm..
cytoplasm
Artifactual degeneration occurs commonly if cells remain
in lavage fluid (saline) for more than a few hours
hours..

Pathological changes
An increase in the PMN % is seen in inflammatory
conditions..
conditions
In septic bronchopneumonia or interstitial
pneumonia the PMN % is usually >20
20%
% (often up
to 90
90%
% with toxic degeneration)
degeneration)..
In allergic pulmonary disease the PMN
eosinophil % and lymphocyte % increase.
increase.

%,

The PMN % increases in proportion to the

severity of the inflammation (up to 15
15%
% in mild
cases, up to 20
20%
% in moderate cases and over
50
50%
% in severe cases)
cases)..

12/6/2011

(Wrights stain x 1000). Fine needle aspirate smear from a

consolidated lung lobe in a 15-year-old cat. This revealed
marked
pyogranulomatous
inflammation.
Numerous
Toxoplasma tachyzoites were found within macrophages
and neutrophils and also free in the background. These
findings, coupled with the radiographic appearance,
suggested
granuloma
formation
secondary
to
toxoplasmosis.

Degenerative changes in PMNs indicate

the possibility of bacterial infection.
infection.
In cases of EIPH numerous hemosiderin
laden macrophages are seen
seen..
Engulfed fungal spores may be seen in
dust allergy cases
cases..

Fine needle aspirate smear from a 12-year-old Cocker

Spaniel with a history of lethargy and mild cough.
Numerous clusters and small sheets of atypical epithelial
cells were present in direct fine needle aspirate smears.
Several acinar structures were found, as shown below.

(Wrights stain x 500). Section of a large adult worm present in a

BAL from a 10-year cat with a 2 week history of coughing. The
morphology of the ova suggested Capillaria infestation.
Eosinophilic inflammation can be seen in the background.
Capillaria is only rarely associated with respiratory disease in
cats.

Bone Marrow Cytology

Indications to perform bone marrow cytology :
1. Investigation of unexplained cytopaenias
a. Unexplained non regenerative anaemia, especially when
persistent and progressive.
b. Unexplained leukopaenia, thrombocytopaenia

2. Investigation of atypical cells in the peripheral blood

a. Immature haematopoietic cells (blasts) raise the possibility of
haematopoietic neoplasia.
Bone marrow evaluation is necessary to determine the source and
number of these cells in the tissue.
b. Persistent, unexplained marked increases in RBC, WBC or
platelet numbers in the peripheral blood.

Illinois Bone Marrow Needle

Bone marrow sampling:

3. Investigation of haemic neoplasia

a.

Differentiation, diagnosis, and staging of

leukaemias and lymphomas.
b. Diagnosis and staging of other neoplasias,
including histiocytic neoplasia, multiple myeloma,
mast cell neoplasia and metastatic carcinoma.
A) Instruments required:
15- to 18-gauge , 1 5 cm long bone marrow
aspiration needle
Several clean microscope slides
5 10 ml syringe
1 ml of a 3 % EDTA/isotonic fluid solution

Bone marrow aspirate from multiple myeloma

showing numerous plasma cells.

In dogs and cats, bone marrow can be aspirated

using sterile technique from the iliac crest, femur or
humerus using an appropriately sized bone marrow
aspiration needle and syringe.

Smear preparation:
Bone marrow degenerates rapidly after collection.
Smears should be prepared immediately after
collection.
Prepare as many smears as possible with the
available marrow.

10

12/6/2011

Intraoral cytology

Abdominal, thoracic, and pericardial effusions

Sample :
Samples from oral cavity collected by scrape,
swab, touch impression or fine needle

Inflammatory cells:
cells:
>85
85%
% neutrophils = suppurative
<50
50%
% macrophages = active
pyogranulomatous
>50
50%
% macrophages = chronic
pyogranulomatous
>10
10%
% eosinophils = eosinophilic

Normally, only a small amount of fluid is present in

the body cavities of animals.
This fluid provides lubrication that allows the
frictionless movement of adjacent organ surfaces
and the body cavity walls.
Effusions
are
the
abnormal
or
increased
accumulation of this fluid in any of the body cavities
that are lined by mesothelial cells.
These include the
abdominal cavities.

thoracic,

pericardial,

and

A cotton swab used to obtain cellular sample from a

mandibular swelling in a 55-month
month--old Labrador Retriever.

Fluid collection
Thoracocentesis:
Thoracocentesis is performed at the seventh or eighth intercostal
space at the level of the costochondral junction, with the patient
standing or in ventral recumbency.
The area is clipped, and the skin is aseptically prepared and infiltrated
with a local anesthetic.
A 1-inch needle is inserted into the pleural space, and a large syringe
can then be used to collect fluid aseptically.

Abdominocentesis

Pericardiocentesis

Abdominocentesis can be performed with the patient in a

standing position or in left lateral recumbency.
An area 1 to 2 cm caudal to the umbilicus and just lateral to
the midline is clipped and aseptically prepared.
The bladder should be emptied before fluid collection. A
needle or catheter is inserted at an angle of 45 to the body
wall.

A portion of the fluid (23 mL) should be placed in a Vacutainer tube

containing EDTA for cell counts, protein analysis, and cytologic
evaluation.

Open-needle abdominocentesis may be more effective than

attaching a needle to a syringe. Therefore, fluid is collected
directly into sterile vials with and without EDTA as described
previously.

A second portion (23 mL) should be placed in a sterile container

without anticoagulant in case aerobic or anaerobic cultures are
required or to assess whether or not clot formation occurs in bloody
fluids.

Also a multifenestrated dialysis catheter may be more

effective in draining fluid than using either a needle or a
butterfly catheter.

A large area from the third to seventh intercostal space is

clipped and aseptically prepared. The site of entry is usually
lateral to the sternum and near the costochondral junction
between the fourth and fifth or fifth and sixth ribs.
The skin and the intercostal muscles located at the site are
infiltrated with a local anesthetic, and a small skin incision is
made.
A 16-gauge, 3.25-inch catheter is inserted at this site at an
angle of 45 until fluid begins to flow. Care should be taken to
avoid the intercostal vessels located at the caudal edge of each
rib.

Total protein
g/dL

Cells per
ml

Cell types

Special
features

Transudate

<2.5

<1000

Mononuclear

Low cellularity

Modified
transudate

2.55.0

1000 8000

Mononuclear

Cell type
varies with
etiology

Nonseptic
exudate

>3.0

>3000

Neutrophils

Nondegenerat
e

Septic
exudate

>3.0

>3000

Neutrophils

Degenerate
neutrophils

Hemorrhagic

>3.0

Variable

Similar to
blood

Erythrophagia
or
hemosiderin in
macrophages

Tumor cells

Neoplastic cell
seen

Fluid analysis
Protein determination
Cell counts

Neoplastic

>2.5

Variable

Pericardiocentesis is a relatively safe procedure and a

technique that is within the capabilities of all small animal
practitioners.
Electrocardiographic (ECG) monitoring is advised during the
procedure, because puncturing the epicardium often results
in ventricular arrhythmias.
Pericardiocentesis is best attempted on the right side, at
the fourth or fifth intercostal space, with the animal
positioned in left lateral recumbency.
An approach from the right side minimizes the risk of
trauma to the lungs and major coronary vessels.
The procedure is usually performed under local anesthesia,
but sedation may be required in some cases.

Classification of effusions
Transudates
Transudates are effusions of low protein and cell
content. They are typically clear and colorless, with
protein concentrations of less than 2.5 g/dL and less
than 1000 nucleated cells per milliliter.
Cytologically,
these
fluids
contain
mostly
mononuclear
cells,
such
as
lymphocytes,
macrophages, and mesothelial cells, with lower
numbers of nondegenerate neutrophils.
Effusions of any kind may irritate the mesothelial
lining of the body cavity, resulting in mesothelial
hyperplasia and eventual sloughing of mesothelium
into the fluid.

11

12/6/2011

Pure transudates most frequently form as a result of

hypoproteinemia from either increased loss or decreased
production of albumin.
Albumin maintains the plasma colloidal osmotic pressure within
the vascular system, preventing leakage of fluid into and
promoting reabsorption of fluid from the extravascular
compartments, such as the body cavities.

1.

2.
a.
b.

Clinical conditions that result in pure transudate formation

include
Decreased production of albumin with liver failure,
malnutrition, maldigestion, or malabsorption.
Increased loss of albumin in
protein-losing nephropathies, including glomerulonephritis,
nephrotic syndrome, and renal amyloidosis.
protein-losing enteropathies, such as lymphangiectasia or
lymphocytic/ plasmacytic enteritis, chronic hemorrhage,
intestinal parasitism, and intestinal neoplasia.

Modified transudates
Its an effusion that occurs by transudative mechanisms, where
vascular fluids leak out of normal or noninflamed vessels (eg,
via increased capillary hydrostatic pressure or lymphatic
obstruction).
Modified transudates result from the leakage of fluid from vessels
carrying high-protein lymphatics or blood, however the fluid is
modified by the addition of protein and/or cells.
Modified transudates color may vary from tan and slightly turbid,
to pink, depending on the etiology. The protein content ranges
from 2.5 to 5.0 g/dL.
In a modified transudate, most of the nucleated cells are
mononuclear cells, either macrophages, lymphocytes, or a
combination of both.
A modified transudate may be a transitory stage of an effusion,
and if the fluid remains in the body cavity long enough, the
protein content and degenerating cells result in chemotaxis of
neutrophils into the area. This alters the classification of these
fluids from a modified transudate to a nonseptic exudate.

Exudates
Exudates are the result of leakage of fluid from abnormal or
altered vasculature. Occurs because of an inflammatory process
or chemotactic stimuli within the body cavity.
The inflammatory process increases serosal and vascular
permeability, resulting in a fluid with elevated protein and
cellularity.
Exudates are further classified as septic or nonseptic depending
on whether or not infectious agents are identified in the fluid.
Exudates may vary in color from white to amber to pink, are
usually turbid. The protein content is usually high (>3 g/dL),
and the cell counts are typically higher than 3000 cells per
milliliter.

Septic exudates
The identification of phagocytized intracellular
organisms, usually bacteria, distinguishes a septic
exudate from a nonseptic one.

The predominant cell type in most septic exudates

is the neutrophil. Many of these cells are
degenerate as evidenced by nuclear karyolysis
(swollen pale nucleus) or karyorrhexis (nuclear
fragmentation).

Most exudates contain a predominant population of

neutrophils, but some inflammatory fluids may
contain a significant (10%) eosinophil component.
These exudates may be specifically termed
eosinophilic effusions.
Causes
of eosinophilic effusions include allergic
hypersensitivity conditions, parasitic diseases (eg,
heartworm
disease,
intestinal
parasites),
pneumothorax,
lung
lobe
torsion,
intestinal
lymphangiectasia,
and
lymphomatoid
granulomatosis.

Many septic exudates occur:

by introducing organisms into the body
cavity via traumatic puncture wounds; bite
wounds
perforation of the intestinal tract;
migrating foreign bodies
ruptured pulmonary, hepatic, or prostatic
abscesses
Pyometra
pneumonia or pleuritis.
Most septic exudates involve bacterial sepsis;
however,
infections
with
Mycoplasma,
rickettsial agents, fungal agents, and
parasites may occur less frequently.

Chylous effusions
Chylous effusions result from leakage of lymphatic fluid into the
thoracic and/ or abdominal cavity.
They are opaque white to pink with cell counts and protein
content similar to modified transudates.
The lymphocytes seen in chylous effusions are small and well
differentiated - contain nuclei that are approximately the size of
erythrocytes with dense dark nuclear chromatin.The cytoplasm is
scant.

Causes of pleural chylous effusions include mediastinal

neoplasms (lymphoma and thymoma), diaphragmatic or
peritoneopericardial hernia (cat), lung torsion, intestinal
lymphangiectasia (dog), mediastinal cryptococcal granuloma, or
idiopathic chylous effusions.

Nonseptic exudates
Clinical conditions, such as
feline infectious peritonitis (FIP)
foreign objects or material in the body cavity
Pancreatitis
bile or urine leakage
neoplasms
torsion of internal organs (eg, lung lobes, liver
lobes, spleen)
inflamed internal organs, or walled-off abscesses
result in a nonseptic exudate.

Neoplastic effusions
Neoplasia is a common cause of effusions in dogs and cats.
Report states that 57% of pericardial effusions and 11% of
peritoneal and pleural effusions in the dog were the result of
neoplasia.
Neoplastic cells
preparations.

are not

always

identifiable

on

cytologic

Neoplastic processes occurring within the body cavities may

result in various types of fluid accumulations, including modified
transudates, exudates, and hemorrhagic effusions.
The term neoplastic effusion should only be used to describe
effusions where a neoplastic cell population has been identified
in the fluid.
Making this determination is difficult, because neoplastic cells
are absent in many effusions caused by neoplasia and reactive
mesothelial cells often have cytologic criteria that mimic
malignancy.

12

12/6/2011

Samples for cytologic evaluation are collected by

fine-needle aspiration, touch impression, or
gentle tissue scraping.
Superficial cutaneous and subcutaneous tissues
such as peripheral lymph nodes are easily
sampled by fine needle aspiration.
Ulcerated masses can be sampled by impression
smears, fine-needle aspiration, or gentle tissue
scraping.
Ultrasound is useful for guiding needle aspiration
biopsies of focal or diffuse neoplastic infiltrations
involving internal organs.

Cytologic Features of Neoplasia

In general, neoplastic cells are large and pleomorphic
in size and shape when compared with normal cells of
the same type.
Benign neoplastic lesions yield cells that are relatively
uniform in size and appearance.
Nuclei have a similar chromatin pattern, and nucleoli
usually are small and regular in outline or
inconspicuous. Rarely do nuclei of benign cells exceed
two to three times the size of homologous red blood
cells in diameter.
There is minimal variation in N:C ratio in cells from
most benign tumors.

CYTOLOGIC INTERPRETATION
Cytologic findings should be correlated with other
clinical and laboratory findings and with information
about tumor incidence, site predilection, and gross
morphology.
Inflammation and Hyperplasia:
Many tumors elicit an inflammatory response that
may mask the presence of neoplastic cells.
Inflammation is characterized by a cells including
neutrophils, lymphocytes, plasma cells, eosinophils,
monocytes, and macrophages.
The presence of inflammation may be problematic in
the cytologic diagnosis of neoplasia because of the
small amount of tissue evaluated and the lack of
architecture to define the demarcation between
inflammation and neoplasia.

Malignant cells exhibit moderate to marked

variation in cell size, referred to as anisocytosis.
Cells from malignant tumors usually are
macrocytic compared to nonneoplastic cells of the
same type.
The N:C ratio often varies markedly from cell to
cell. For most tumors, the N:C ratio increases
with malignant transformation.
Malignant cells from some tumors appear to be at
different stages of differentiation and may exhibit
asynchrony between nuclear and cytoplasmic
maturation.

Inflammatory reactions often result in hyperplasia

of surrounding tissues. Cells from hyperplastic
tissue resemble normal cells except they appear
more immature.
Cytologic characteristics of hyperplastic cells
include large nuclei with poorly condensed
chromatin and prominent nucleoli. Cytoplasm often
is basophilic.
Hyperplastic cells have a relatively constant nuclear
to- cytoplasmic (N:C) ratio (nuclear size compared
with the amount of cytoplasm present), an
important feature in distinguishing hyperplastic
from neoplastic cells.

The nuclear membrane appear irregularly

thickened, angular, or indented. Some malignant
cells show nuclear molding around adjacent
nuclei or cytoplasmic vacuoles.
Nucleoli may be large (macronucleoli) in
malignant cells. Nucleoli greater than 5 microns
in diameter are highly suggestive of malignancy.
Nucleoli also may be irregularly shaped, and
more than one nucleolus per cell may be present.
Anisonucleoliosis (variation in nucleolar
occurs more frequently in malignant cells.

size)

Thank you

13