2 The uterus Uterus is lined by simple columnar ciliated cells cervix Fundus of the uterusBody of the uteruscervixUterus is lined by simple columnar ciliated cells
3 The cervix Anatomy Histology vagina Stratified squamous non-keratinzed epithelium rich in glycogenEctocervixEndocervixPseudo-stratified columnar ciliated epithelium, rich in mucus secretionCervical canal
4 The vagina Function: protection of upper genital tract. Thickness: Child: thin (few cells) and atrophyMenopause: thin and atrophyPuberty (reproductive period): thick (more than one layer, 7-10 layers).
5 Cytology of normal female genital tract 1- Epithelial cells:Stratified squamous epitheliumColumnar epitheliumEndometriumGlandular endometrial cellsStromal endometrial cells:Superficial cellsDeep cells
8 Glandular endometrial cells Physiologically: they are present as normal cells in the following conditions, in routine vaginal smearMenstrual flow ( early 2-3 days).Early pregnancyAbortionPost-partum periodIn case of intra-uterine device at the end of menstrual cycle.
10 2- Non-epithelial cells Smooth muscle cells: normally not present except in deep traumatic scrapping of ulcerative lesion as in polyp, fibroma, and abortion.RBCs Plasma cellsHistocytes (macrophages)Sperms YeastTrichomanos vaginalis
11 Inflammatory cells usually posterior fornix or cervical smears BacterialVaginosisPolymorphsDiplococciInflammatory changes in epithelial cellsPresence of polymorphsIntra-cellular diplococci
12 Candida spores-hyphae Dust mitesTrichomonasCandida spores-hyphaePresence of micro-organismsTrichomonasCandidaBacteriaMites
14 Cervical – vaginal smears Several possibilitiesCervical scrape taken from the squamo-columnar junction; the level of the SC junction varies with sexual maturityPosterior fornix aspirationLateral vaginal wall [upper third] scrape
15 Squamo-Columnar Junction Junction of pink cervical skin and red endocervical canalInherently unstableKey portion of the cervix to sampleMost likely site of dysplasiaIn obtaining the Pap smear, it is important to sample the "Squamo-columnar Junction." This is the circular area right at the opening of the cervix where the pink, smooth skin of the cervix meets the fiery-red, fragile, mucous-producing lining of the cervical canal. If there is going to be a problem with cancer or precancerous changes, it is this area that is most likely to be effected. This area is also known as the SQJ, or transition zone.
16 Vaginal SpeculumThe vaginal speculum consists of two blades. When closed, the speculum is inserted into the vagina. Then the blades are opened, revealing the cervix at the end of the vagina. Thin-bladed Pederson speculums are most comfortable for most women. For women who have had babies, the wider Grave’s speculum may be needed for good visualization of the vagina and cervix.
17 Ayers Spatula Concave end to fit the cervix Convex end for vaginal wall and vaginal pool scrapingsThe Ayers spatula is specially designed for obtaining Pap smears. The concave end (curving inward) fits against the cervix, while the convex end (curving outward) is used for scraping vaginal lesions or sampling the "vaginal pool," the collection of vaginal secretions just below the cervix.The spatula is made of either wood or plastic. Both give very satisfactory results.
18 Cytobrush Insert ~ 2 cm (until brush is fully inside canal) Rotate only 180 degrees (otherwise will cause bleeding)Push the cytobrush into the canal, no deeper than the length of the brush (1.5 cm cm). Rotate the brush 180 degrees (half a circle) and pull the cytobrush straight out. Don't keep spinning the brush round and round or you will cause bleeding. Even the degree rotation may cause a little bleeding but usually it doesn't.
19 Squamous epithelial cells Keratinized type: skin, valvaNon-keratinized: vagina, ectocervix- Basal cells: usually not seen unless there is sever ulceration with destruction of whole thickness of epithelium: deep basophilic- Parabasal cells- Intermediate cells- Superfacial cells- Non-nucleated cells
20 Differentiating feature of normal squamous cells ParabasalIntermediateSuperficialCell formationSingle cells or sheetSingle cells or clumpDiameter15 – 25 umum33 – 55 umNucleusVesicular with finely granular chromatin umVesicular with finely granular chromatin 8 – 10 umPyknotic or absent 4 – 6 umCytoplasmOval, thick, and dark basophilicThin, flat or folded, polyhydral, pale basophilicPolyhydral, thin, flat, eosinophilic
21 Feature of cell maturation NucleusCytoplasmBasal cellsReticular chromatinGreenish blue (basophilic)Parabasal and intermediate cellsGranular chromatinYellowSuperficial cellsPyknotic chromatinRed (acidophilic)
22 Make Pap Smear As thin as possible Properly labeled Label the slide with pencil on the frosted end.Two slides may be made, one for the spatula and one for the brush (“two-slide” technique). Alternatively, a single slide may be used (the “one-slide” technique) in which the brush is spread on one half the slide and the spatula is used on the other half. Both techniques give good results.
23 Spray with Fixative Within 10-15 seconds Allow to fully dry before packagingCytologic FixativeAllow the slides to dry completely before placing them in the Pap smear container. Once dry and packaged, it is best to send them out promptly for interpretation. When operational circumstances disallow prompt sending of the slides, they can be held for weeks to months without significant loss of readability.Make sure the slides are properly labeled and that important clinical information is included with the requisition. Telling the cytologist that the patient has had a hysterectomy will save considerable amounts of time in evaluating the smear.For women who have had a hysterectomy, Pap smears are obtained by using the convex end of the Ayers spatula, scraping it horizontally across the top of the vagina. Then the cytobrush is used to reach into the the right and left top corners of the vagina.
24 Causes of unsatisfactory smear Air drying smear before fixation ---- less differentiation between cytoplasm and nucleus.Inadequate fixationUncleaned slidesIncorrect staining timeDryness during staining procedures
25 Characters of unsatisfactory smear ThickPoorly spreadBlood smear with clotting