Universität zu Köln

Grafen, Markus ORCID: 0000-0002-6941-9235, Hofmann, Thurid R., Scheel, Andreas H., Beck, Julia, Emmert, Alexander, Kueffer, Stefan, Danner, Bernhard C., Schutz, Ekkehard, Buettner, Reinhardt, Ostendorf, Andreas ORCID: 0000-0002-3601-6392, Stroebel, Philipp and Bohnenberger, Hanibal (2017). Optimized expression-based microdissection of formalin-fixed lung cancer tissue. Lab. Invest., 97 (7). S. 863 - 873. NEW YORK: NATURE PUBLISHING GROUP. ISSN 1530-0307

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Abstract

Analysis of specific DNA alterations in precision medicine of tumors is crucially important for molecular targeted treatments. Lung cancer is a prototypic example and one of the leading causes of cancer-related deaths worldwide. One major technical problem of detecting DNA alterations in tissue samples is cellular heterogeneity, that is, mixture of tumor and normal cells. Microdissection is an important tool to enrich tumor cells from heterogeneous tissue samples. However, conventional laser capture microdissection has several disadvantages like user-dependent selection of regions of interest (ROI), high costs for dissection systems and long processing times. ROI selection in expression-based microdissection (xMD) directly relies on cancer cell-specific immunostaining. Whole-slide irradiation leads to localized energy absorption at the sites of most intensive staining and melting of a membrane covering the slide, so that tumor cells can be isolated by removing the complete membrane. In this study, we optimized xMD of lung cancer tissue by enhancing staining intensity of tumor cell-specific immunostaining and processing of the stained samples. This optimized procedure did not alter DNA quality and resulted in enrichment of mutated EGFR DNA from lung adenocarcinoma specimens after xMD. We here also introduce a quality control protocol based on digital whole-slide scanning and image analysis before and after xMD to quantify selectivity and efficiency of the procedure. In summary, this study provides a workflow for xMD, adapted and tested for lung cancer tissue that can be used for lung tumor cell dissection before diagnostic or investigatory analyses.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Grafen, Markus UNSPECIFIED orcid.org/0000-0002-6941-9235 UNSPECIFIED Hofmann, Thurid R. UNSPECIFIED UNSPECIFIED UNSPECIFIED Scheel, Andreas H. UNSPECIFIED UNSPECIFIED UNSPECIFIED Beck, Julia UNSPECIFIED UNSPECIFIED UNSPECIFIED Emmert, Alexander UNSPECIFIED UNSPECIFIED UNSPECIFIED Kueffer, Stefan UNSPECIFIED UNSPECIFIED UNSPECIFIED Danner, Bernhard C. UNSPECIFIED UNSPECIFIED UNSPECIFIED Schutz, Ekkehard UNSPECIFIED UNSPECIFIED UNSPECIFIED Buettner, Reinhardt UNSPECIFIED UNSPECIFIED UNSPECIFIED Ostendorf, Andreas UNSPECIFIED orcid.org/0000-0002-3601-6392 UNSPECIFIED Stroebel, Philipp UNSPECIFIED UNSPECIFIED UNSPECIFIED Bohnenberger, Hanibal UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-226055
DOI:	10.1038/labinvest.2017.31
Journal or Publication Title:	Lab. Invest.
Volume:	97
Number:	7
Page Range:	S. 863 - 873
Date:	2017
Publisher:	NATURE PUBLISHING GROUP
Place of Publication:	NEW YORK
ISSN:	1530-0307
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language LASER-CAPTURE MICRODISSECTION; PROMOTER METHYLATION; MOLECULAR ANALYSIS; CELLS; SAMPLES; GENE Multiple languages Medicine, Research & Experimental; Pathology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/22605