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Cytology and

Cytological
Techniques
Clinical Pathology
Cytology
The microscopic examination
of cells.
Generally refers primarily to
cells exfoliated from tissues,
lesions, and internal
organ/tumor cells.
A very valuable diagnostic tool.
Is inexpensive
Is quick and easy
Involves little or no risk to the
patient
Cytology Continued

Must be able to identify normal cells from


abnormal cells, and inflammatory from
non-inflammatory cells

Disadvantage may be that some tumors


do not exfoliate cells well and therefore
may not provide and adequate sample to
examine.
Cytologic Interpretation

May be able to diagnose


Identify the disease process
Help form a prognosis
May determine what diagnostic
procedures should be performed next
May help with therapy options
Cytologic Techniques

Fine Needle Aspirate (FNA)


Fluid Aspiration-
Thoracocentesis/Abdominocentesis
Solid mass imprinting
Vaginal wall technique
Cerebrospinal (CSF) Fluid Analysis
Synovial Fluid Analysis
Nasal Flush
General Collection
Techniques
When possible prepare several smears
Use stained and unstained techniques
May use a variety of stains
Use clean, dry slides
Scrapings

Done on freshly cut surfaces


Scrap lesion/tissue with clean scalpel
blade
Place material collected on a slide and
spread
Advantage: May collect more cells
Disadvantage: More difficult to collect
and only able to collect superficial lesions
Imprints

May be prepared from external lesions


(ulcers)
May be prepared from tissues excised
during surgery or necropsy.
Easy to collect
Disadvantage: May only collect few cells
and may contain contamination
Solid Mass imprints
Cut mass in half
Blot dry
Need to remove blood/tissue
fluid from surface
Use sterile gauze or other
absorbent material
Excess blood/fluid inhibits cells
from spreading and assuming
normal size and shape
Touch the slide to the blotted
surface
Stain
Fine Needle Aspirates
Preferred method of obtaining samples from
masses.
Avoids superficial contamination
Very little risk to patient
Less complications to internal organs than core
biopsy techniques
Implantation of malignant cells along the aspiration
tract is extremely rare
Disadvantage: May not get a good sample
because using just a small needle.
Fine Needle Aspirate

2 techniques
Aspiration
Collect with 22-25 gauge needle
Use 3-12 ml syringe
Need slides
Non-aspiration
FNA Aspiration
Technique
Hold mass/lymph node firmly
Introduce the needle with syringe attached into the mass
Apply strong negative pressure by withdrawing the plunger to
about 2/3 -3/4 of the volume.
Do several times in same area or redirect needle.
Stop negative pressure and remove needle from mass
Remove needle from syringe and air is drawn up into syringe
Sample that is in hub of needle is expelled onto slide by rapidly
depressing the plunger
Hold needle close to slide, if too far away will result in small
droplets that dry rapidly before smear technique may be done.
FNA Non-Aspiration
Technique
Works best for small
masses that are difficult
to aspirate.
Works well for highly
vascular tissues
Using a needle only,
move rapidly back and
forth (stabbing motion).
Withdraw needle and
place syringe with air to
force onto slide.
Preparation of smears
from aspirates
Squash prep method
Needle spread method
Blood smear method
Squash Preparation
With experience, can yield excellent cytologic smears
Aspirated material is placed on the center of the slide
A second slide is placed over the sample to form a
cross.
Carefully slide apart from first slide (Put down on and
pick up to move).
Do not place excessive downward pressure to the first
slide because will cause distorted ruptured cells
The weight of the spreader slide is sufficient to
adequately spread the cells.
Needle Spread Method

Spread aspirate on the slide with tip of


needle.
Pull sample out into several projections
(starfish appearance).
Blood Smear Technique
Use if material is thick or
fluid
After material is expelled
on slide, second slide is
held at 30-40angle.
Second slide is pulled
backward until it contacts
the fluid
Rapidly move forward like
a blood smear.
Common Problems with
FNA
Few or no cells obtained
Some lesions do not exfoliate cells well.
The needle may miss the site of the lesion
Timid collection
Inadequate negative pressure
Blood contamination
Using too large needle gauge
Prolonged aspiration
Failure to blot if doing imprint
Common Problems with
Preparation
Poorly prepared slides due to thick or high cell
numbers
Allowing material to dry on slide before squash
prep or other smear technique.
If a large amount of material is present, spread
between two slides
May have to do 4-5 slides form the same site in
order to get valuable diagnostic sample.
Staining Slides
Diff-quik, Wrights, Geimsa
Papanicolau stains-
used in human Ob/gyn exams. Stains nucleus and
nuclear material better.
New Methylene Blue stain
Air dry these slides, do not heat fix.
Use clean slides (make sure no lint on slide)
Stain immediately after air drying
Take care not to touch the surface of the slide
or smear at any time.
Medical Terminology
Hypertrophy-an increase in cell size and/or
functional activity in response to a stimulus.
Hyperplasia- increase in cell numbers, via
increased mitotic activity, in response to a
stimulus.
Neoplasia- increase in cell growth and
multiplication that is not dependent on an
external stimulus.
Metaplasia- a reversible process in which one
mature cell type is replaced by another mature
cell type (adaptive response to a stimulus)
Medical Terminology
Continued
Dysplasia- reversible, irregular, atypical,
proliferative cellular changes in response to
irritation or inflammation.
Anaplasia- A lack of differentiation of tissue
cells
Less differentiated cells in a tumor is more
malignant
Chromatin pattern- the microscopic pattern of
nuclear chromatin (the chromatin pattern
coarsens as malignant potential increases)