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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  • 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


  • 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


  • 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)