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Recommended for : 1. Detection of malignant cells 2. Detection of precancerous (precursor) lesions 3. Assessment of female hormonal status 4. Determination of genetic (phenotypic) sex 5. Detection of the presence of infx microorganisms
SPECIMENS FOR EXAMINATION 1. Peritoneal, pericardial and pleural fluids 2. CSF 3. Nipple discharge 4. Bronchial brushings / washings 5. Sputum 6. Gastric washings 7. Urine sediment 8. Prostatic secretions / fluid 9. Cervicovaginal (Paps) smear
COLLECTION OF SPECIMEN
Collection Procedure: Use an unlubricated speculum (saline or warm water may be used). After visualization of the cervix is accomplished, insert the sampling device (e.g. cytobrush) into the endocervical canal and rotate one complete turn. Withdraw the cytobrush and spread the collected material quickly and evenly onto the slide opposite the frosted end.
COLLECTION OF SPECIMEN
COLLECTION OF SPECIMEN
COLLECTION OF SPECIMEN
COLLECTION OF SPECIMEN
WHY FIX? Exfoliated cells decompose rapidly which may destroy cellular and nuclear details, in turn will give inadequate results for diagnosis. COMMON FIXATIVES 1.Equal parts of 95% ethanol and ether 2.95% ethanol 3.Carnoys fluids 4.Equal parts of tertiary butyl alcohol and 1 part 95% ETOH 5.Schaudinns Fluid Sat. Aq. Hg2Cl, Abs. ETOH, HAc 6.Methanol for dried films
PAPANICOULAOU or Paps Smear ADVANTAGE: - Transparent blue staining of cytoplasm is observed - Excellent nuclear staining - Color range is predictable and of great value in identification of cells DISADVANTAGE: - Procedure is lengthy and complicated - Does not give accurate acidophilic index
RESULTS : Epithelial cytoplasm either pink-red to orange to (eosinophilic) greenish blueto blue- gray (basophilic) Vesicular nucleus blue Pyknotic nucleus dark blue to black Bacteria (L. acidophilus) dark blue Fungal mycelia violet Trichomonas vaginalis pale greenish blue blob of cytoplasm
vesicular nucleus blue pyknotic nucleus dark blue to black
Vaginal cytology:
Vaginal epithelium Responsive to sex steroids, particularly estrogen and progesterone Undergoes predictable changes through the cycle Responds to changes in blood concentrations of ovarian hormones. Rising levels of estrogen cause the vaginal epithelium to become "cornified" The surface cells become large and flattened, with small or absent nuclei.
Vaginal cytology:
Vaginal cytology A type of endocrine assay. Provides a convenient means of assaying changes in estrogen & progesterone levels Changes in the morphology of desquamated vaginal epithelial cells
Vaginal cytology:
Vaginal smears may be taken regularly and often. Hormonal changes are best mirrored in the upper third of the vagina. They can also be taken from the lateral walls because their more accessible and less likely to be contaminated by cellular debris or discharge.
Superficials
Intermediates
Parabasals
SUPERFICIAL CELLS
Large (30-60u) Polyhedral flat cells Cytoplasm: may be acidophilic or basophilic Presence of small dark pyknotic nuclei (less than 6u)
SUPERFICIAL CELLS Anucleate cells are abnormal which may be derived from: 1. Smear contamination by the cells from the vulva 2. Epidermization of the vagina or cervix resulting from prolapse 3. Leukoplakia of the cervix 4. Ruptured membranes in pregnant women 5. Marked hyper-estrinism Medium large (20-30u) Polyhedral or elongated
INTERMEDIATE CELLS
PARABASAL CELLS
Has a translucent basophilic cytoplasm (due to glycogen accumulation) Cytoplasm stains deep blue or blue gray + cell membrane
NAVICULAR CELLS
ENDOCERVICAL CELLS
Boat-shaped intermediate cells with a strong tendency to fold and curl their edges. Expression of excess progesterone effect Found in the latter half of menstrual cycle, during pregnancy, menopause May also be found as a result of abnormal androgen stimulation, either endogenous or exogenous Tall columnar epithelial cells with basal nuclei Occurs in groups and strips of three or more cells ( picket fence configuration) Cytoplasm : basophilic with whispy mucin vacuole
ENDOMETRIAL CELLS
Found during menstruation period ( in groups) seen up to 10 days from LMP Endometrial stromal cells : seen in tight clusters of small, oval dark cells Glandular cells: slightly larger Nucleus: small and moderately dark Cytoplasm: basophilic and maybe vacuolated
Gram (+) slender rod bacteria Predominant organism of the vaginal normal flora: establishes the low pH that inhibits the growth of pathogens Stains pale blue to lavender Energy is obtained by the fermentation of glycogen derived from disintegrating epithelial cells Numerous in the luteal phase and during pregnancy
PYKNOTIC INDEX (P.I) karyo-pyknotic index; percentage of cells having shrunken, dark, small (less than 6) nuclei.
ADEQUACY
General Categorization:
Negative for Intraepithelial lesion or malignant cell Benign Cellular Changes Epithelial cell abnormalites
Descriptive Diagnosis:
Infections / Radiation effects Atypical squamous / glandular cells of unknown significance Low grade squamous intraepithelial lesion High grade squamous intraepithelial lesion Squamous Cell Carcinoma Adenocarcinoma Others
Pap smear specimens are considered satisfactory for interpretation if there are: Adequate numbers of well-visualized squamous cells present Adequate numbers of well-visualized endocervical cells or squamous metaplastic cells (from the transformation zone). Properly labeled specimens
ADEQUACY
Specimen Unsatisfactory
ADEQUACY
Specimens to which the following conditions apply will be rejected: 1. Specimen is submitted without a requisition. 2. Specimen is not labeled with the patient name. 3. The patient name (or other identifying information) on the specimen and requisition do not correspond. 4. The specimen is labeled appropriately but the requisition is not labeled. 5. The specimen slide(s) is (are) irreparably broken. 6. Specimen is submitted from an unauthorized source.
Pap smear specimens are considered unsatisfactory for interpretation if there are: Inadequate numbers of well-visualized squamous cells present Inadequate numbers of well-visualized endocervical cells or squamous metaplastic cells (from the transformation zone). More than 75% of the cells obscured by blood or inflammation Improperly labeled specimens Usually, these smears are recommended for repeat sampling.
High grade squamous intraepithelial lesion (HSIL) (encompassing: moderate and severe dysplasia, CIS, CIN 2 and CIN 3) - with features suspicious for invasion (if invasion is suspected)
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