EXFOLIATIVE CYTOLOGY AND HORMONAL CYTOLOGY

• Exfoliative cytology


  Branch of general cytology which deals with the microscopic study of cells desquamated, removed or shed from epithelial surfaces
• Purposes of exfoliative cytology

  • Detect cancerous / malignant conditions
  • Detect asymptomatic cancer in women or precancerous cervical lesions
  • Determination of genetic sex
  • Detection of possible infection
• Possible infections detected by exfoliative cytology

  • HPV
  • Trichomoniasis
  • Yeast infection (e.g candidiasis)
• Hormonal cytology


  Evaluate female hormonal status in case of sterility
• Gynecological specimens for exfoliative cytology

  • Cervico vaginal smear a.ka PAP smear
• Non-gynecological fluid specimens for exfoliative cytology

  • Sputum
  • Smears of urine sediments
  • Endometrial and endocervical smears
  • Prostatic and breast secretions
  • Pleural and peritoneal fluids
  • Gastric and bronchial secretions
  • CSF
• Preparation methods for non-gynecologic specimens

  • Smears
  • Cell block
  • Cytospin preparation
• Respiratory tract specimens


  • Bronchoalveolar Lavage
  • Bronchial washings
  • Bronchial brushings
  • Sputum
• Sputum


  Deep cough, 3 samples for 3 consecutive mornings, check for alveolar macrophages to determine if it is sputum or saliva
• Sputum processing


  Immediate fixation because cells degenerate immediately, use Saccomano fluid as fixative
• Gastric secretions/aspirates/lavage


  Immediate transport to lab required, no delay beyond 30 minutes, fasting for at least 8 hours prior to collection
• Breast secretion/nipple discharge


  Low diagnostic yield for breast cancer diagnosis, discharge during and post lactation may be normal, discharge is usually benign and may be due to duct ectasia & papilloma
• Breast Smear preparation techniques

  • Pull apart technique
  • Fix using spray fix or 95% Isopropyl Alcohol (IPA)
• Peritoneal, pleural, pericardial fluid


  Accumulation of these fluids is usually indicative of pathologic process, have a tendency to clot so add heparin to prevent jelly clot
• Urinary tract specimens


  • Voided urine
  • Catheterized urine
  • Washing from bladder or renal pelvis
• Urine specimen collection


  First voided must be discarded, second collection is preferred, if delay then refrigerate but do not add preservatives, 50 mL specimen must be collected
• Body cavity fluids (CSF, ascitic fluid)

  Specimen must be submitted fresh, formalin or alcohol should not be used, at least 1 mL of CSF is required
• Smear preparation techniques


  • Streaking
  • Spreading
  • Pull-apart
  • Touch Preparation/ Impression smear
• Specimens tested in histopathology

  • Autopsy specimens
  • Biopsy specimen
  • Cytology specimen
• Centrifugation of non-gynecological/fluid specimens

  Carried out at 1,000 rpm for 1 minute, decant supernatant and smear sediments, use extra sediments for cell block
• Cell block preparation methods

  • Direct Filtration
  • Plasma Thrombin method
  • Carbowax
• Specimen preparation for microscopy

  Smears should be prepared from fresh material and fixed (10-15 min) while moist to prevent drying
• Specimens that require adhesive

  • Urinary sediments
  • Bronchial Lavage
  • Concentrated Sputum
  • Specimen with Trypsin
• Adhesives used in cytology

  • Pooled serum or plasma
  • Celloidin ether alcohol
  • Leuconostoc culture: Gram (+) cocci
  • APES 3 aminopropyltriethoxysilane
• Reason why egg albumin cannot be used as adhesive

  Intense staining by the counterstain used in PAPs, counterstain stains the cells and adhesive
• Most common tissue adhesive

  Mayer's egg albumin
• Fixatives used in cytology

  • 95% ethanol ether
  • 95% ethanol
  • Spray fixative
• Saccomano preservative


  Combination of 50% alcohol and 20% carbowax
• Fixatives for different specimens

  • Cervical/vaginal smears: 95% Ethanol and Ether, 95% Ethanol

  Sputum and bronchopulmonary: 95% Ethyl alcohol, Cough directly on a container with Brasil's fixative

  Pleural and peritoneal fluids: Brasil's fixative, 50% Alcohol

  Gastric specimen: 95% Ethanol

  All types of Effusion: 50% Alcohol

  Sputum: 70% Alcohol

  Urine, Gastric and Bronchial Aspirates: 95% Alcohol
• Other fixatives


  • Carnoy's fluid, equal parts of tertiary butyl; alcohol and 1 part of 95% ethanol, Schaudinn's fluid
• Most common tissue fixative

  10% Buffered Neutral Formalin
• Papanicolau stain


  Gold Standard stain for Cytology
• Routine tissue processing stain

  H&E staining, Hematoxylin - nuclear dye, Eosin - cytoplasmic dye
• Components of Pap's stain

  • Harris Hematoxylin (nuclear stain)

  OG6 (counterstain; cytoplasm stain for superficial cells)

  EA 50 (counterstain; cytoplasm stain of both parabasal and intermediate cells)
• Staining results


  • Vesicular Nucleus: Blue

  Pyknotic nucleus: Dark blue to black

  OG6: Orange with a hint of green

  EA36-50: Olive green with a hint of brown and red

  Bacteria: Dark blue

  Mycelia: Violet

  T. vaginalis: Pale greenish blue
• Purposes of Pap's smear


  • Screening test for detection of cervical cancer

  Hormonal Cytology
• Conventional Pap's smear


  OB will collect specimen using Ayre's spatula and smear on the slide, resulting in overlapping and crowded cells
• Ary butyl


  Alcohol and 1 part of 95% ethanol
• Schaudinn's fluid


  Tissue fixative
• Carnoy's fluid


  Best for bloody specimens