DIAGNOSTIC CYTOLOGY

• Microscopic examination of cells from different body sites for

diagnostic purposes
• Sample must include representative material
• Method used to process the specimen must give rapid results
• Consistency and reliability are most important in cytological
interpretation
• Specimen for cytologic examination:
• Exfoliative cytology
• Fine needle aspiration (FNA)
• Body fluids
EXFOLIATIVE
CYTOLOGY

Presented by: Krisha Imee

Anjanelle Mesia
EXFOLIATIVE
CYTOLOGY
microscopic study of cells that
have been shed, desquamated
or scraped off from lining
epithelium and mucosal
surfaces
 Anton Van Leeuwenhoek

19th Century
• Rudolf Virchow : cell theory and doctrines of
cellular pathology
• Pouchet: described the cytologic population
HISTORICAL changes of the vagina in women and animals
1935
BACKGROUND
• Dudgeon: systematic investigations done on
the cytodiagnosis of cancer of the lungs
1943
• Papaniculao and Traut: “Diagnosis of
Uterine Cancer by the Vaginal Smear”
PURPOSES OF EXFOLIATIVE
CYTOLOGY

1 2 3 4 5
assessing malignant detecting assessing female determining genetic determining presence
or cancerous asymptomatic cancer hormonal activity (in sex of possible infections
conditions in women case of
sterility/endocrine
disorders)
ACCURATE CYTOLOGIC INTERPRETATION OF
CELLULAR MATERIALS IS DEPENDENT UPON:
method of specimen collection

fixation and fixatives

preservation of fluid specimens prior to processing

preparation of materials for microscopic examination

staining and mounting of the cell sample

COLLECTION
OF SPECIMEN
• vaginal scrapings
: scraping/swab

• endocervical and
endometrial
scrapings : swab,
curettage
COLLECTION OF SPECIMEN
• prostatic secretions : • bronchial aspirate/sputum :
prostatic massage deep cough or bronchoscopy
COLLECTION OF SPECIMEN : serosal fluids
• pleural effusion : • pericardial effusion :
thoracentesis pericardiocentesis
COLLECTION OF
SPECIMEN : serosal
fluids

• peritoneal fluid/
ascites : paracentesis
COLLECTION OF SPECIMEN
• gastric and duodenal fluids : • cerebrospinal fluid: lumbar
nasogastric tube aspiration puncture (spinal tap)
COLLECTION OF SPECIMEN
• bone marrow aspirate : bone • urinary sediments : urine
marrow tap collection or catheterization
Smear Preparation
 Take note:
• Exfoliated cells-decompose rapidly
➢ prepare smears immediately after removal from
the body and fixed
➢ if smears cannot be prepared immediately, place in
FIXATION proper fixative and refrigerated until it can be
prepared
AND • Minimum fixation time varies : 15 mins
• Recommendation:
FIXATIVES ➢ smears should be left in fixative for a minimum of 1
hr before staining
• Smears may be left for weeks or months
provided that the container is covered lightly
preventing evaporation of the solution
COMMONLY USED FIXATIVES
1. Alcohol-ether (Ether- alcohol)
-equal parts of 95% alcohol and ether
-best fixative for cytological smears

2. 95% Ethyl Alcohol

-universal fixative, most widely used
-does not contain volatile and flammable ether
-may be used for all smears prepared at the side of the patient(vaginal and cervical
smears, prostatic smears, breast smears and aspiration biopsy smears)
-used for final fixation of all smears
COMMONLY USED FIXATIVES
COMMONLY USED FIXATIVES
3. Carnoy’s Fluid
➢useful for bloody specimes : hemolyze rbc’s
➢shrinkage of epithelial cells is greater than in 95% ethanol
➢staining time in hematoxylin must be reduced to prevent overstaining
➢place smears in Carnoy’s fixative for 3-5 mins until the sediment is lost if smears
remain in Carnoy’s fixative for longer than 15 mins
➢fixative must be prepared fresh when needed and discarded after each use

4. Coating Fixatives
-hairsprays (containing high alcohol content with a minimum of oil or
lanolin or oil
➢ have dual action
▪ fix cells when dry
▪ form a thin protective coating over the smear
*not recommended for bloody smears (clumping of erythrocytes)
COMMONLY USED FIXATIVES
-COATING FIXATIVES-

Take note:
• the can should be shaken well prior to each use
• should be applied immediately to fresh smears
• distance between nozzle of the spray and the slide affects quality of the cytologic
detail
=10-12 in (25-30 cm)
• If the spray is held too close :
-cells may be dislodged by the force of the spray
-evaporation of the spray vehicle may freeze and irreversibly
damage cells
-artifact may occur
• spray must be smooth and steady
 1. Smears should be placed into the fixative immediately.
2. Place each smear in fixative by a single uninterrupted
motion, avoiding rippled smear material.
3. Avoid striking the bottom of the container forcefully to
prevent dislodging the materials from the slide.
4. Attach a paper clip to the identified end of the slide.
5. Specimens with a high mucus content maybe preserved
PRECAUTIONS for 12-24 hrs. if refrigerated
IN FIXATION: 6. Specimens with high protein content maybe preserved
for 24-48 hrs. if refrigerated
7. Specimens with low mucus or protein content will
endure only 1-2 hrs. delay even if refrigerated
8. Specimens with a low pH must be collected on ice and be
prepared within minutes of collection to prevent cellular
destruction by HCl