1. Anatomy and Histology of female Genital Tract

2. The uterus Uterus is lined by simple columnar ciliated cells cervix
Fundus of the uterus
Body of the uterus
cervix
Uterus is lined by simple columnar ciliated cells

3. The cervix Anatomy Histology vagina
Stratified squamous non-keratinzed epithelium rich in glycogen
Ectocervix
Endocervix
Pseudo-stratified columnar ciliated epithelium, rich in mucus secretion
Cervical canal

4. The vagina Function: protection of upper genital tract. Thickness:
Child: thin (few cells) and atrophy
Menopause: thin and atrophy
Puberty (reproductive period): thick (more than one layer, 7-10 layers).

5. Cytology of normal female genital tract
1- Epithelial cells:
Stratified squamous epithelium
Columnar epithelium
Endometrium
Glandular endometrial cells
Stromal endometrial cells:
Superficial cells
Deep cells

6. Squamous Epithelium

7. Superficial
Intermediate
Parabasal
Basal

8. Glandular endometrial cells
Physiologically: they are present as normal cells in the following conditions, in routine vaginal smear
Menstrual flow ( early 2-3 days).
Early pregnancy
Abortion
Post-partum period
In case of intra-uterine device at the end of menstrual cycle.

9. Glandular endometrial cells
Pathologically: present in endometrial lesions as:
Polyp
Endometritis
Submucous fibroid
Endometrial hyperplasia
Adeno-carcinoma

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 cells
Histocytes (macrophages)
Sperms Yeast
Trichomanos vaginalis

11. Inflammatory cells usually posterior fornix or cervical smears
Bacterial
Vaginosis
Polymorphs
Diplococci
Inflammatory changes in epithelial cells
Presence of polymorphs
Intra-cellular diplococci

12. Candida spores-hyphae
Dust mites
Trichomonas
Candida spores-hyphae
Presence of micro-organisms
Trichomonas
Candida
Bacteria
Mites

13. Methods of collecting endometrial samples
Endometrial aspiration
Endometrial lavage
Endometrial scraping
Endometrial biopsy

14. Cervical – vaginal smears
Several possibilities
Cervical scrape taken from the squamo-columnar junction; the level of the SC junction varies with sexual maturity
Posterior fornix aspiration
Lateral vaginal wall [upper third] scrape

15. Squamo-Columnar Junction
Junction of pink cervical skin and red endocervical canal
Inherently unstable
Key portion of the cervix to sample
Most likely site of dysplasia
In 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 Speculum
The 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 scrapings
The 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, valva
Non-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
Parabasal
Intermediate
Superficial
Cell formation
Single cells or sheet
Single cells or clump
Diameter
15 – 25 um um
33 – 55 um
Nucleus
Vesicular with finely granular chromatin um
Vesicular with finely granular chromatin 8 – 10 um
Pyknotic or absent 4 – 6 um
Cytoplasm
Oval, thick, and dark basophilic
Thin, flat or folded, polyhydral, pale basophilic
Polyhydral, thin, flat, eosinophilic

21. Feature of cell maturation
Nucleus
Cytoplasm
Basal cells
Reticular chromatin
Greenish blue (basophilic)
Parabasal and intermediate cells
Granular chromatin
Yellow
Superficial cells
Pyknotic chromatin
Red (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 packaging
Cytologic Fixative
Allow 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 fixation
Uncleaned slides
Incorrect staining time
Dryness during staining procedures

25. Characters of unsatisfactory smear
Thick
Poorly spread
Blood smear with clotting

26. Thank you