Cytology o Vaginal scrape

 Study of the microscopic appearance of cells for diagnostic purposes o Lateral vaginal scrape
 Can be used as a screening tool for healthy individuals at risk of a particular o Four quadrant vaginal scrape
disease o Vulvar scrape
 Exfoliative vs FNA cytology  Specimen collection
o Exfoliative – cells desquamated from epithelial surfaces o A bivalve speculum of appropriate size is gently inserted into the patient’s
 Spontaneous shedding vagina
 Physically removed from epithelial and mucous membranes o A cervical brush/broom/spatula is inserted in the endocervical canal
o FNA – fine needle aspiration of palpable and non-palpable masses o For conventional smears
 Applications of Exfoliative Cytology  The brush sample should be rolled over the slide followed by
o Detection of malignant cells in body fluids (for staging cancers) immediate fixation
o Detection of precancerous cervical lesions (cervicovaginal or Pap smear) o For liquid-based preparations
o Assessment of female hormonal status in case of sterility and endocrine  Cells are rinsed into a liquid collection media containing fixatives
disorders  Ensures the capture of an entire sample from the collection
 devices
 o Liquid-based preparations
o For determination of genetic sex  alternative to conventional cervicovaginal smears
o Detection of infectious agents  Improved preparation that minimizes cell overlap for better
 Gynecological vs Non-gynecological cytology identification of abnormal cells
o Gyn cytology  ThinPrep technique:
 specimens from the female genital tract  SurePath technique:
o Non-gyn cytology  Gynecological Cytology: Staining
 Specimens from all other body sites o Papanicolau method
 Cytology: Fixation o Nuclear stain
 Specimen should be fixed immediately for optimal cell preservation  Hematoxylin
o 95% ethanol o Cytoplasmic stains – 2 counterstains
o Equal parts of 95% ethanol and ether  Orange G
 If smears from effusions cannot be made immediately, place in:  Eosin Azure
o 50% alcohol  Components:
o Saccomano preservative o Staining technique
 If fluid specimen is enough for cytocentrifugation: 1. Fix in 95% EtOH
o Centrifuge at 2000 rpm for 2 mins, decant supernatant and smear the 2. Primary stain with hematoxylin
sediment 3. Differentiate with acid alcohol then wash with water
o Prepare at least 2 cytocentrifuged smears and a cell block (similar to 4. Blue in ammonia water then wash with water
tissue processing) 5. Counterstain with OG-6
6. Wash with 2 changes of 95% EtOH
Gynecological Cytology 7. Counterstain with EA-50
 Transformation zone/T-zone 8. Dehydrate (ascending grades of alcohol)
o Endocervical-ectocervical junction 9. Clear with xylene
o Where majority of cervical CA and precancerous lesions of the cervix arise 10. Mount with resinous media
 Cytologic collection and preparation  Bethesda System Categories for Specimen Adequacy
o For conventional pap smears o Satisfactory for evaluation
o Endocervical brush  A satisfactory squamous component must be present.
  Note the presence/absence of endocervical/transformation zone  Perimenopausal:
component. o After childbearing age
 Obscuring elements (inflammation, blood, drying artifact, other)  Postmenopausal:
may be mentioned if 50% to 75% of epithelial cells are obscured.  Teleatrophy:
o Unsatisfactory for evaluation  Gynecological Cytology: Normal Cell Components
 Specimen rejected/not processed because [specify reason]. o Neutrophils
Reasons may include: o RBCs
 Lack of patient identification o Lactobacillus acidophilus (Doderlein bacilli)
 Unacceptable specimen (e.g., slide broken beyond  Gram positive bacillus that is a part of the vaginal normal flora
repair) o Leptothrix spp
 Specimen processed and examined, but unsatisfactory for  Long, thin, filamentous, hair-like bacilli that are normal
evaluation of an epithelial abnormality because [specify reason]. commensals and become prolific if vaginal pH increases
Reasons may include: o Endocervical cells
 Insufficient squamous component  Columnar epithelial cells that are part of the normal lining of the
 Obscuring elements covering more than 75% of endocervical canal
epithelial cells o Endometrial cells
 Cytohormonal Maturation Index (CHMI)  Small epithelial cells from the shedding of the endometrial lining
o Assesses ovarian hormonal function or due to a proliferating endometrial pathology
 Estrogen and Progesterone  Gynecological Cytology: Abnormal Cell Components
 Shows the predominant hormone of the woman at the time of o Candida albicans
collection of the smears  A budding yeast that form a branching pseudohyphae that
o Correlated with the age of the patient and her last menstrual period spears clusters of epithelial cells
o Numerical expression representing the relative proportion of the 3 o Trichomonas vaginalis
vaginal cell types:  Sexually-transmitted pear-shaped organism
 Parabasals, Intermediates, Superficials o Gardnerella vaginalis
 CHMI = P / I / S  Tiny pleomorphic coccobacilli that clings to the surface of the
o Parabasal cells cytoplasm of epithelial cells (clue cells)
 Round to oval cells with small dense basophilic cytoplasm o Koilocytes
 Absence of both estrogen and progesterone  Squamous epithelial cells that show HPV cytopathic effects
o Intermediate cells o HSV-II
 Polyhedral or elongated cells with basophilic cytoplasm  Herpes simplex virus
 Influenced by increased progesterone o High grade squamous intraepithelial lesions (HSIL)
o Superficial cells  Encompassing severe dysplasia and carcinoma in-situ
 Polygonal squamous cells with pale, pink-staining cytoplasm and o Invasive squamous cell carcinoma
dark pyknotic nuclei  Most common form of cervical malignancy
 Influenced by increased estrogen  Bethesda System for Reporting Cervical Cytology
 CHMI Results o Negative for intraepithelial lesion or malignancy (NILM)
o Premenarche:  Organisms
o Childbearing age  Trichomonas vaginalis
 Ovulation:  Fungal organisms morphologically consistent with
 Menstruation: Candida species
 Pregnancy:  Shift in flora suggestive of bacterial vaginosis
 Postpartum:  Bacteria morphologically consistent with Actinomyces
  Cellular changes consistent with herpes simplex virus o Causes:
 Other non-neoplastic findings  Benign breast lesion such as duct ectasia and papilloma
 Reactive cellular changes associated with: inflammation  Endocrine problems
(includes typical repair); radiation; intrauterine  Detection of malignant cells
contraceptive device (IUD)  Urinary Tract Specimen
 Glandular cells status post hysterectomy o Principle: Diagnosis of malignancy, usually of urothelial origin
 Atrophy o Specimen
o Epithelial cell abnormalities  Voided urine (second morning)
 Squamous cell  Catheterized specimen
 Atypical squamous cells (ASC)  Washings from bladder or renal pelvis
o Of undetermined significance (ASC-US)  Body Cavity Effusions
o Cannot exclude HSIL (ASC-H) o Accumulation of fluids within the body cavities usually indicate a
 Low-grade squamous intraepithelial lesion (LSIL) pathologic process
 High-grade squamous intraepithelial lesion (HSIL) o Principle: Presence of malignant cells  metastasis
 Squamous cell carcinoma (SQC) o Specimen:
 Glandular cell  Cytopreparation
 Atypical glandular cells (AGC) (specify if endocervical, o Cell suspensions
endometrial, or not otherwise specified)  For body cavity effusions, CSF, urine, watery lavages
 AGC, favor neoplastic (specify if endocervical or not  Cells are viable up to 4 days at ref temp
otherwise specified)  Standard technique: cytocentrifugation then slides are stained
 Endocervical adenocarcinoma in situ (AIS) with Pap stain
 Adenocarcinoma (specify if endocervical, endometrial,
extrauterine, or not otherwise specified) Fine Needle Aspiration Cytology
o Other: Includes sarcoma, malignant lymphoma, others  FNA of superficial masses and deeply seated lesions
 Specimen collection for non-palpable masses
Non-gynecological Cytology o Aspirated under fluoroscopy, computed tomography, ultrasound or other
 Respiratory Tract Specimens radiologic techniques
o Principle: obtained to exclude the possibility of malignancy or infectious  Specimen collection for palpable masses
agents o breast, thyroid soft tissue and lymph nodes
o Specimen: 1. Palpate the target lesion, sterilize the overlying skin
 Sputum 2. Fix the lesion with one hand between fingers
 Fixative: Saccomanno fluid 3. Introduce the needle (22-23 gauge) and then aspirate
 bronchoalveolar lavage (BAL)  Slide preparation
 bronchial washing (BW) o First few drops from the tip of the needle has the most diagnostic
 bronchial brushing (BB) material
 Gastrointestinal Specimens o Prepare about 4 slides using wedge technique or pull-apart technique
o Principle: Done to exclude the possibility of malignant tumors o Rinse needle in a preservative (Saccomano) and send to the lab
o Specimen:  for cytocentrifugation and/or cell block
o delay of more than 1/2 hour before fixation  digestion of cells   Slide fixation
specimen unsatisfactory for evaluation o 95% alcohol or spray fixative
 Smears of Breast Secretion  Stain
o Low diagnostic yield for diagnosis of breast carcinoma o Same with Papanicolau method
o Specimen: Nipple discharge