1. Diagnostic Cytopathology & Significance of Biopsy Investigation Jaroslava Dušková Inst. Pathol.,1st Med. Faculty, Charles Univ. Prague https://www1.lf1.cuni.cz/~jdusk/

2. Morphological Diagnostic Methods v Clinical v Pathological

3. Morphological Diagnostic Methods v Clinical: v macroscopy of lesions v visible with the naked eye v invisible with the naked eye - IMAGING (X-ray, sonography, scintigraphy, endoscopy, CT,…) v magnifying glass - colposcopy

4. Morphological Diagnostic Methods v Pathological v macroscopy v microscopy v ultrastructure v IMAGING

5. Morphological Diagnostic Methods v Pathological v macroscopy of lesions v autopsy report v biopsy description v cytology material description

6. Morphological Diagnostic Methods v Pathological v microscopy v cytology v minibiopsy - cytoblock v histology

7. Cytology (FNAB) u often both first and final dg. method u outpatient low cost procedure u done by an experienced (cyto)pathologist surprisingly effective u has some limits (!)

8. CYTOLOGY IS VERY EFFICIENT IN EARLY NEOPLASM DIAGNOSTICS

9. ESPECIALLY, IF CLINICO- MORPHOLOGICAL COMMUNICATION WORKS

10. Expectations Clinician from his pathologist: v confirmation of neoplasm dg. v nosological classification v grading, staging v prognosis v reaction to the therapy v recidive recognition

11. Expectations Pathologist from his clinician: v information: local.,size, duration, former dg. a treated neo, clin. dg. v diagnostic material acquisition v correct interpretation of the pathologist´s report

12. Getting Cytology Material u surface – smeared, brushed, scraped u cavities – punctured, aspirated u deep solid lesions - aspirated

13. Processing Cytology Material u smears u cytospins, cytosedimentation u cytoblock

14. Minibiopsy - Cytoblock from FNAB u advantage of easy material taking together with more tissue architecture information u histology &, immunohistochemistry methods available u multilayered tissue phragments readable

15. Cytology Material Staining u gynecology smears - polychrome u other materials –MGG, HE, polychrome, all other methods u cytoblock – multiple methods

16. Goals of Cytological Investigation u Screening – detection of symptomless lesions u Diagnosis of pathological lesions found v introductory (followed by histol.) v final

17. Gynecological oncologic cytology laboratory investigation standard Authors: MUDr Alena Beková, MIAC MUDr Pavel Tretiník, MIAC Oponents: doc. MUDr J. Dušková, CSc,FIAC MUDr Eva Svobodová

18. Cytology - Evaluation u staining and evaluation – minutes u Bethesda system : –material quality and quantity –group diagnosis –dg. as close to histology as possible –recommendation

19. Bethesda System 2001

20. Suitable for Evaluation u without limits u limited by… v non processed v processed but limited for evaluation of squamous cell abnormalities due to…..

21. General Categorisation u negative for intraepith. lesion or malignancy u intraepith. lesion or malignancy squamous or glandular u other pathology endometral cells in women over 40 yrs

22. Interpretation u negative for intraepithelial lesion or malignancy u Microorganisms v Trichomonas v mycosis vs. candidosis v shift - bact. vaginitis v bacteria Actinomyces like v cell changes of HSV type

23. Interpretation u negative for intraepithelial lesion or malignancy v Other non-neoplastic changes v reactive v inflammation (+ repair) v IUD v atrophy

24. Bethesda - cervical cytology classification 1. normal 2. benign cellular changes 3. ASCUS 4. L SIL 5. H SIL 6. Atypiae of glandular cells 7. susp. adenoca

25. Interpretation u epithelial abnormalities v squamous v ASC-US v ASC-H v LSIL CIN 1, HPV v HSIL CIN2, CIN3, CIS v susp. invasion v squamous ca

26. Interpretation u epithelial abnormalities v squamous v ASC-US v ASC-H v LSIL CIN 1, HPV v HSIL CIN2, CIN3, CIS v susp. invasion v squamous ca

27. Bethesda System 2001

28. Cytology getting sample u needle 0.6-0.8mm u min. 2 punctions v aspiration v nonaspiration – reduction of the blood content u cyst: evacuate and aspirate with the second punction the periphery u fluid: whole volume for cytology

29. Peroperation Biopsy u dg. during minutes u morphological artefacts (combination with cytology) u limited extensity of investigation u limited time

30. „Classical“ Biopsy (formol paraffin technique ) u simple may be done in two days u immuno reactions two days more u further sectioning two days more u oncol. dg. - WHO classification –typing, grading, staging –prognostic factors

31. Biopsy u any tissue removed from a living patient is a COMPULSORY subject to u do not follow the tendency to discard „unimportant“ materials (nevi, tonsillae, uteri, intestine, endometrial curretage…..) !

32. Biopsy u do not crush small samples taking imprint cytology !!! u adequate amount of fixation solution (sample: fixative = min. 1:10) u wide neck bottle tightly closed for transport but openable without rough violence u flat sample – spread and tighten on a bearing socket u mark discrete suspicious lesion with a stitch before fixation

33. CYTOLOGY LOVE IT or LEAVE IT L. Cardozzo