Exfoliative Cytology

Exfoliative Cytology Branch of general pathology that studies cells that have been desquamated from the epithelial surface Exfoliative cytology is usually recommended for the following: ① Assessing malignant and cancerous condition ② Detection of asymptomatic cancer in women (vaginal cytology) ③ Assessment of female hormonal activity in case of sterility and endocrine disorders ④ Determination of genetic sex ⑤ Determination for possible infection Specimen for examination: ① Vaginal smears ② Endometrial and endocervical smears ③ Prostatic and breast secretions ④ Gastric and bronchial secretions ⑤ Pleural and peritoneal fluids ⑥ Sputum ⑦ Smears of urine sediments ⑧ CSF Specimens require addition of an adhesive agent: ① Urinary sediments ② Bronchial lavage specimen ③ Specimen that utilizes proteolytic enzymes during processing: a) Trypsin b) Concentrated sputum c) Enzymatic lavage specimen from the GI tract Characteristics of Adhesive Agent: ① Permeable to both fixative and stain ② Must not retain stain Egg albumin is not recommended as an adhesive agent because the basic light green counter stain of Papanicolaou method intensively stains it

Example of good adhesive agents: ① Pooled human serum or plasma ② Celloidin either alcohol ③ Leuconostoc culture Fixation:  Minimum fixing time is 1 hour but smears can be left for 15 – 20 min  Best fixative seems to be ether alcohol  95% ethanol is used alone in most lab due to the flammability and volatility of ether  Common fixatives: ① Equal parts of 95% ethyl alcohol and ether ② 95% ethyl alcohol ③ Carnoy’s fl uid  Other fixatives: ① 3 parts butyl alcohol and 1 part 95% ethyl alcohol

② Schaudinn’s fluid ③ Methyl alcohol (dried blood film) Bleaching section for restaining:  Decolorized old section must be restained  Soak the slide in xylene until coverslip slides off or gently heat the slide  Steps: ① Remove cover slip ② Water ③ 0.5 potassium permanganate (510 min) ④ Running tap water ⑤ 5%oxalic acid (5 min or turn white) ⑥ Running tap water ⑦ Restain Smear Preparation: Four methods for preparing cytological examination: ① Streaking

② Spreading ③ Pull apart ④ Touch or impression smear

smears

for

Staining (papanicolaou method “pap’s smear”)

③ Lower layer or para basal: a) b) c) d)

Staining method of choice for Exfoliative cytology

Advantages of pap’s smear: ① Transparent blue staining of cytoplasm ② Excellent nuclear detain ③ Color range is predictable and of great value in identification and classification ④ Comparing cellular appearances

CHMI   

3 types of papanicolaou staining method: ① Harris Hematoxylin Staining solution ② OG 6 stain ③ EA 50 Stain or EA 36 (eosin azure) Criteria for malignant cell Nuclear change: ① Inc. N/C ratio ② Nuclear pleomorphism ③ Hyper chromasia ④ Irregular chromatin condensation ⑤ Abnormalities in size and number of form of nucleoli ⑥ Multinucleation Cytologic change: ① Excess acidophile ② Abnormal vaculation ③ Abnormal inclusion 3 layers ① Upper layer or superficial: a) High estrogen (shift to the right) b) Dark pyknotic nucleus c) Cytoplasm is flat plus trans d) Polygonal or polyhedral

② Middle layer or intermediate: a) High Progesterone (shift to the left) -> puberty or lactating b) Vesicular nucleus c) Polygonal or elongated d) Basophilic cytoplasm e) Folding effect of cytoplasm (navicular cells)

Round or oval Fried egg or sunny side up Youngest layer High in natural menopause, Surgical operation and post partum period (after child birth)

Para basal / Intermediate / Superficial 100/0/0 is menopause 0/5/95 is normal healthy female

Systems of reporting gynecologic cytology Class system:  No longer used  Manner of reporting: ① Negative for malignant cell ② Atypical cells present ③ Suspicious but not conclusive of malignant cells ④ Strongly suggestive of malignant cells ⑤ Conclusive (positive) for malignant cells The Bethesda System (TBS)  National Cancer institute  TBS report includes: ① Specimen adequacy ② General categorization ③ Descriptive diagnosis  Reported as: ① Satisfactory ② Satisfactory but limited to ③ Unsatisfactory