Microsatellite Analysis of Hereditary Nonpolyposis Colorectal Cancer-Associated Colorectal Adenomas by Laser-Assisted Microdissection Giuseppe Giuffrè,

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Microsatellite Analysis of Hereditary Nonpolyposis Colorectal Cancer-Associated Colorectal Adenomas by Laser-Assisted Microdissection Giuseppe Giuffrè, Annegret Müller, Thomas Brodegger, Tina Bocker-Edmonston, Johannes Gebert, Matthias Kloor, Wolfgang Dietmaier, Frank Kullmann, Reinhard Büttner, Giovanni Tuccari, Josef Rüschoff The Journal of Molecular Diagnostics Volume 7, Issue 2, Pages 160-170 (May 2005) DOI: 10.1016/S1525-1578(10)60542-9 Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Laser microdissection. A: A periodic acid-Schiff-stained section used to identify the degree of dysplasia. B, C, E: The laser cut line surrounds the dysplastic cells (B); these cells were harvested in the cap of a microtube (C, E). D: The tissue section after laser microdissection: DNA contamination by stromal and inflammatory cells surrounding the desired cells was avoided. The Journal of Molecular Diagnostics 2005 7, 160-170DOI: (10.1016/S1525-1578(10)60542-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Microsatellite status after manual and laser-guided dissection indicating the cases with a changed MSI level. The Journal of Molecular Diagnostics 2005 7, 160-170DOI: (10.1016/S1525-1578(10)60542-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Carcinoma arising in an adenoma in a case with partial loss of MSH2 in the adenoma. A: Focus of MSH2 expression in the adenoma (->, retained MSH2 immunostaining; AD, adenoma; CA, carcinoma; N, normal mucosa; *, lymphoid follicle as internal positive staining control). B: Enlarged inset from A with focal MSH2 staining and C shows the expression of MLH1 within the same area. D and E: MSI at BAT40 (D) and BAT25 (E) of MSH2-negative cells (D: lane 1, normal; lanes 2 and 3, adenomatous tissue without MSH2; lanes 4 and 5, with preserved MSH2 expression; E: lane 1, normal; lane 2, adenomatous tissue with MSH2 staining; and lanes 3 and 4, without MSH2 staining). The Journal of Molecular Diagnostics 2005 7, 160-170DOI: (10.1016/S1525-1578(10)60542-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 A known HNPCC patient with synchronous adenomas. One adenoma showed nuclear staining for MLH1 and MSH2 (A) whereas the other revealed a loss of MLH1 (B). Corresponding MSI analysis disclosed instabilities at BAT25 (top set) and BAT 26 (bottom set) for the one (B) and no instabilities for the other (A) adenoma (lanes 1 and 4, normal control). The Journal of Molecular Diagnostics 2005 7, 160-170DOI: (10.1016/S1525-1578(10)60542-9) Copyright © 2005 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions