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 30-40˚angle.
 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, Wright’s, 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)