Cytology and Cytology and Cytological Cytological TechniquesTechniques

Clinical PathologyClinical Pathology

CytologyCytology

The microscopic examination The microscopic examination of cells.of cells. Generally refers primarily to Generally refers primarily to

cells exfoliated from tissues, cells exfoliated from tissues, lesions, and internal lesions, and internal organ/tumor cells.organ/tumor cells.

A very valuable diagnostic tool.A very valuable diagnostic tool. Is inexpensiveIs inexpensive Is quick and easyIs quick and easy Involves little or no risk to the Involves little or no risk to the

patientpatient

Cytology ContinuedCytology Continued

Must be able to identify normal cells from Must be able to identify normal cells from abnormal cells, and inflammatory from abnormal cells, and inflammatory from non-inflammatory cellsnon-inflammatory cells

Disadvantage may be that some tumors Disadvantage may be that some tumors do not exfoliate cells well and therefore do not exfoliate cells well and therefore may not provide and adequate sample to may not provide and adequate sample to examine.examine.

Cytologic InterpretationCytologic Interpretation

May be able to diagnose May be able to diagnose Identify the disease processIdentify the disease process Help form a prognosisHelp form a prognosis May determine what diagnostic May determine what diagnostic

procedures should be performed nextprocedures should be performed next May help with therapy optionsMay help with therapy options

Cytologic TechniquesCytologic Techniques

Fine Needle Aspirate (FNA)Fine Needle Aspirate (FNA) Fluid Aspiration- Fluid Aspiration-

Thoracocentesis/AbdominocentesisThoracocentesis/Abdominocentesis Solid mass imprintingSolid mass imprinting Vaginal wall techniqueVaginal wall technique Cerebrospinal (CSF) Fluid AnalysisCerebrospinal (CSF) Fluid Analysis Synovial Fluid AnalysisSynovial Fluid Analysis Nasal FlushNasal Flush

General Collection General Collection TechniquesTechniques

When possible prepare several smearsWhen possible prepare several smears Use stained and unstained techniquesUse stained and unstained techniques

May use a variety of stainsMay use a variety of stains

Use clean, dry slidesUse clean, dry slides

ScrapingsScrapings

Done on freshly cut surfacesDone on freshly cut surfaces Scrap lesion/tissue with clean scalpel Scrap lesion/tissue with clean scalpel

bladeblade Place material collected on a slide and Place material collected on a slide and

spreadspread Advantage: May collect more cellsAdvantage: May collect more cells Disadvantage: More difficult to collect Disadvantage: More difficult to collect

and only able to collect superficial lesionsand only able to collect superficial lesions

ImprintsImprints

May be prepared from external lesions May be prepared from external lesions (ulcers)(ulcers)

May be prepared from tissues excised May be prepared from tissues excised during surgery or necropsy.during surgery or necropsy.

Easy to collectEasy to collect Disadvantage: May only collect few cells Disadvantage: May only collect few cells

and may contain contaminationand may contain contamination

Solid Mass imprintsSolid Mass imprints

Cut mass in halfCut mass in half Blot dryBlot dry

Need to remove blood/tissue Need to remove blood/tissue fluid from surfacefluid from surface Use sterile gauze or other Use sterile gauze or other

absorbent materialabsorbent material Excess blood/fluid inhibits cells Excess blood/fluid inhibits cells

from spreading and assuming from spreading and assuming normal size and shapenormal size and shape

Touch the slide to the blotted Touch the slide to the blotted surfacesurface

StainStain

Fine Needle AspiratesFine Needle Aspirates

Preferred method of obtaining samples from Preferred method of obtaining samples from masses.masses.

Avoids superficial contaminationAvoids superficial contamination Very little risk to patientVery little risk to patient

Less complications to internal organs than core Less complications to internal organs than core biopsy techniquesbiopsy techniques

Implantation of malignant cells along the aspiration Implantation of malignant cells along the aspiration tract is extremely raretract is extremely rare

Disadvantage: May not get a good sample Disadvantage: May not get a good sample because using just a small needle.because using just a small needle.

Fine Needle AspirateFine Needle Aspirate

2 techniques2 techniques AspirationAspiration

Collect with 22-25 gauge needleCollect with 22-25 gauge needle Use 3-12 ml syringeUse 3-12 ml syringe Need slidesNeed slides

Non-aspirationNon-aspiration

FNA Aspiration TechniqueFNA Aspiration Technique

Hold mass/lymph node firmlyHold mass/lymph node firmly Introduce the needle with syringe attached into the massIntroduce the needle with syringe attached into the mass Apply strong negative pressure by withdrawing the plunger to Apply strong negative pressure by withdrawing the plunger to

about 2/3 -3/4 of the volume.about 2/3 -3/4 of the volume. Do several times in same area or redirect needle.Do several times in same area or redirect needle. Stop negative pressure and remove needle from massStop negative pressure and remove needle from mass Remove needle from syringe and air is drawn up into syringeRemove needle from syringe and air is drawn up into syringe Sample that is in hub of needle is expelled onto slide by rapidly Sample that is in hub of needle is expelled onto slide by rapidly

depressing the plungerdepressing the plunger Hold needle close to slide, if too far away will result in small Hold needle close to slide, if too far away will result in small

droplets that dry rapidly before smear technique may be done.droplets that dry rapidly before smear technique may be done.

FNA Non-Aspiration FNA Non-Aspiration TechniqueTechnique

Works best for small Works best for small masses that are difficult masses that are difficult to aspirate.to aspirate.

Works well for highly Works well for highly vascular tissuesvascular tissues

Using a needle only, Using a needle only, move rapidly back and move rapidly back and forth (stabbing motion).forth (stabbing motion).

Withdraw needle and Withdraw needle and place syringe with air to place syringe with air to force onto slide.force onto slide.

Preparation of smears Preparation of smears from aspiratesfrom aspirates

Squash prep methodSquash prep method Needle spread methodNeedle spread method Blood smear methodBlood smear method

Squash PreparationSquash Preparation

With experience, can yield excellent cytologic smearsWith experience, can yield excellent cytologic smears Aspirated material is placed on the center of the slideAspirated material is placed on the center of the slide A second slide is placed over the sample to form a A second slide is placed over the sample to form a

cross.cross. Carefully slide apart from first slide (Put down on and Carefully slide apart from first slide (Put down on and

pick up to move).pick up to move). Do not place excessive downward pressure to the first Do not place excessive downward pressure to the first

slide because will cause distorted ruptured cellsslide because will cause distorted ruptured cells The weight of the spreader slide is sufficient to The weight of the spreader slide is sufficient to

adequately spread the cells. adequately spread the cells.

Needle Spread MethodNeedle Spread Method

Spread aspirate on the slide with tip of Spread aspirate on the slide with tip of needle.needle.

Pull sample out into several projections Pull sample out into several projections (starfish appearance). (starfish appearance).

Blood Smear TechniqueBlood Smear Technique

Use if material is thick or Use if material is thick or fluidfluid

After material is expelled After material is expelled on slide, second slide is on slide, second slide is held at 30-40held at 30-40˚angle.˚angle.

Second slide is pulled Second slide is pulled backward until it contacts backward until it contacts the fluidthe fluid

Rapidly move forward like Rapidly move forward like a blood smear.a blood smear.

Common Problems with Common Problems with FNAFNA

Few or no cells obtainedFew or no cells obtained Some lesions do not exfoliate cells well.Some lesions do not exfoliate cells well. The needle may miss the site of the lesionThe needle may miss the site of the lesion Timid collectionTimid collection Inadequate negative pressureInadequate negative pressure

Blood contaminationBlood contamination Using too large needle gaugeUsing too large needle gauge Prolonged aspirationProlonged aspiration Failure to blot if doing imprintFailure to blot if doing imprint

Common Problems with Common Problems with PreparationPreparation

Poorly prepared slides due to thick or high cell Poorly prepared slides due to thick or high cell numbersnumbers

Allowing material to dry on slide before squash Allowing material to dry on slide before squash prep or other smear technique.prep or other smear technique.

If a large amount of material is present, spread If a large amount of material is present, spread between two slidesbetween two slides

May have to do 4-5 slides form the same site in May have to do 4-5 slides form the same site in order to get valuable diagnostic sample. order to get valuable diagnostic sample.

Staining SlidesStaining Slides

Diff-quik, Wright’s, GeimsaDiff-quik, Wright’s, Geimsa Papanicolau stains- Papanicolau stains-

used in human Ob/gyn exams. Stains nucleus and used in human Ob/gyn exams. Stains nucleus and nuclear material better. nuclear material better.

New Methylene Blue stainNew Methylene Blue stain Air dry these slides, do not heat fix.Air dry these slides, do not heat fix. Use clean slides (make sure no lint on slide)Use clean slides (make sure no lint on slide) Stain immediately after air dryingStain immediately after air drying Take care not to touch the surface of the slide Take care not to touch the surface of the slide

or smear at any time.or smear at any time.

Medical TerminologyMedical Terminology

HypertrophyHypertrophy-an increase in cell size and/or -an increase in cell size and/or functional activity in response to a stimulus.functional activity in response to a stimulus.

HyperplasiaHyperplasia- increase in cell numbers, via - increase in cell numbers, via increased mitotic activity, in response to a increased mitotic activity, in response to a stimulus.stimulus.

NeoplasiaNeoplasia- increase in cell growth and - increase in cell growth and multiplication that is not dependent on an multiplication that is not dependent on an external stimulus.external stimulus.

MetaplasiaMetaplasia- a reversible process in which one - a reversible process in which one mature cell type is replaced by another mature mature cell type is replaced by another mature cell type (adaptive response to a stimulus)cell type (adaptive response to a stimulus)

Medical Terminology Medical Terminology ContinuedContinued

DysplasiaDysplasia- reversible, irregular, atypical, - reversible, irregular, atypical, proliferative cellular changes in response to proliferative cellular changes in response to irritation or inflammation.irritation or inflammation.

AnaplasiaAnaplasia- A lack of differentiation of tissue - A lack of differentiation of tissue cellscells Less differentiated cells in a tumor is more Less differentiated cells in a tumor is more

malignantmalignant Chromatin patternChromatin pattern- the microscopic pattern of - the microscopic pattern of

nuclear chromatin (the chromatin pattern nuclear chromatin (the chromatin pattern coarsens as malignant potential increases)coarsens as malignant potential increases)