Hologic Proprietary © 2012 ThinPrep ® Pap Test Diagnostic Challenges and Differential Diagnoses.

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Hologic Proprietary © 2012 ThinPrep ® Pap Test Diagnostic Challenges and Differential Diagnoses

Hologic Proprietary © 2012 ThinPrep ® Characteristics Wet Fixation –enhanced cytoplasmic and nuclear detail –variability in nuclear staining Cell Size –proportionately smaller –single cells more prominent –cells may round up in solution e.g.. adenocarcinoma Smear Pattern –Cellular material not pulled out in mucous –Mechanical artifacts eliminated Specimen Background –Cellular debris may appear clumped

Hologic Proprietary © 2012 Differential Diagnoses Endocervical Adenocarcinoma vs. Poorly Differentiated Squamous Cell Carcinoma (SCC) Endocervical Adenocarcinoma vs. Endometrial Adenocarcinoma Endometrial Adenocarcinoma vs. Small Cell Squamous Carcinoma Adenocarcinoma-in-situ vs. Tubal Metaplasia HSIL vs. Single Endometrial Cells HSIL vs. Immature Squamous Metaplasia Poorly Differentiated Squamous Cell Carcinoma vs. Repair

Hologic Proprietary © 2012 Poorly Differentiated Squamous Cell Carcinoma vs. Endocervical Adenocarcinoma Poorly Differentiated SCC 2D sheets and single cells Ragged group edges Dense, homogenous cytoplasm Pleomorphism, irregular nuclear shapes and sizes Irregular chromatin clumping Nucleoli variable in shape, size, number and position Endocervical Adenocarcinoma 3D cell groupings Common group borders Delicate, foamy cytoplasm Enlarged nuclei, commonly round/oval Parachromatin clearing Round, central, single or multiple macronucleoli

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Hologic Proprietary © 2012 Endocervical Adenocarcinoma vs. Endometrial Adenocarcinoma Endocervical Adenocarcinoma Abundant abnormal material –directly scraped Well preserved material Cells and cell groupings generally larger in size Abundant, foamy cytoplasm, occasionally columnar shaped AIS precursor with endocervical architecture may be seen Endometrial Adenocarcinoma Isolated abnormal groups –cell shedding Variable preservation of cells Cells and cell groupings generally smaller in size Scant, cyanophilic cytoplasm with occasional conspicuous vacuoles Mature hormonal pattern, watery transudate may be seen

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Hologic Proprietary © 2012 Endometrial Adenocarcinoma vs. Small Cell SCC Endometrial Adenocarcinoma Isolated abnormal groups cell shedding Occasional clusters, fewer single cells Scant, vacuolated cytoplasm Eccentrically located nuclei Parachromatin clearing Round, central, single or multiple macronucleoli Small Cell SCC Abundant abnormal material directly scraped ++ Single cells, clusters Dense, homogenous cytoplasm Centrally located nuclei Irregular chromatin clumping Prominent, irregular nucleoli

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Hologic Proprietary © 2012 Adenocarcinoma In Situ vs. Tubal Metaplasia Adenocarcinoma In Situ Crowded sheets & strips Feathering & pseudostratification Relative hyperchromasia Uniformly stippled chromatin Nuclear size and shape variation Block-like nucleoli Mitoses and apoptotic bodies Tubal Metaplasia Cilia and/or terminal bars reliably identified Crowding without overlap, lacking depth of focus Evenly distributed chromatin Cytoplasm dense and cuboidal Nucleoli absent Nuclear membrane irregularities and thickening absent Occasional elongate nuclear forms

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Hologic Proprietary © 2012 HSIL vs. Endometrial Cells HSIL Sheets, syncitia; thick plaques rather than 3D ball-like clusters Hyperchromasia Irregular nuclear membranes Single cells have centrally located nuclei Dense homogenous cytoplasm Endometrial Cells 3D ball-like clusters and small, single cells Relative hyperchromasia Regular nuclear membranes Single cells with eccentrically located nuclei Scant basophilic cytoplasm with cytoplasmic “blebs”

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Hologic Proprietary © 2012 HSIL vs. Immature Squamous Metaplasia HSIL Single cells, clusters, and thickened plaques Variable hyperchromasia Irregular nuclear membranes Coarse clumped chromatin Variation in nuclear size within a cluster Immature Squamous Metaplasia Single cells & cobblestone sheets Normochromatic Regular nuclear membranes Evenly distributed chromatin with pinpoint chromocenters Nuclear size & shape consistent

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Hologic Proprietary © 2012 Repair vs. Poorly Differentiated SCC Poorly Differentiated SCC Poorly cohesive sheets and single cells Irregular nuclear shapes & sizes Thickened nuclear membranes Irregular chromatin clumping Nucleoli variable in shape, size, number and position Some nuclei display nucleoli, some nuclei do not Repair (typical) Cohesive sheets Significant nuclear size variation but typically round/oval in shape Thin, well defined nuclear membranes More open chromatin pattern with minimal variability Centrally located macronucleoli with smooth, round contours Virtually all nuclei display nucleoli

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Hologic Proprietary © 2012 Trademark Statement CytoLyt, Hologic, PreservCyt, ThinPrep, and UroCyte are registered trademarks of Hologic, Inc. and/or its subsidiaries in the United States. CytoLyt, Hologic, PreservCyt, ThinPrep, UroCyte and associated logos are trademarks of Hologic, Inc. and/or its subsidiaries in other countries. All other trademarks are the property of their respective owners.

Hologic Proprietary © 2012 Any Questions? Part No Rev. 001