cytology and cytological techniques
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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

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cytology
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
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
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
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
General Collection Techniques
  • When possible prepare several smears
  • Use stained and unstained techniques
    • May use a variety of stains
  • Use clean, dry slides
scrapings
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
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
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
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
Fine Needle Aspirate
  • 2 techniques
    • Aspiration
      • Collect with 22-25 gauge needle
      • Use 3-12 ml syringe
      • Need slides
    • Non-aspiration
fna aspiration technique
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
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
Preparation of smears from aspirates
  • Squash prep method
  • Needle spread method
  • Blood smear method
squash preparation
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
Needle Spread Method
  • Spread aspirate on the slide with tip of needle.
  • Pull sample out into several projections (starfish appearance).
blood smear technique
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
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
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
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
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
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)
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