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18 results on '"Giesen, Charlotte"'

5. Automatic single cell segmentation on highly multiplexed tissue images

Author

Schüffler Peter J, Schapiro Denis, Giesen Charlotte, Wang Hao A O, Bodenmiller Bernd, and Buhmann Joachim M

Subjects
Diagnostic Imaging, ComputingMethodologies_PATTERNRECOGNITION, ComputingMethodologies_IMAGEPROCESSINGANDCOMPUTERVISION, Humans, Breast Neoplasms, Female, Single-Cell Analysis, Flow Cytometry, Immunohistochemistry
Abstract

The combination of mass cytometry and immunohistochemistry (IHC) enables new histopathological imaging methods in which dozens of proteins and protein modifications can be visualized simultaneously in a single tissue section. The power of multiplexing combined with spatial information and quantification was recently illustrated on breast cancer tissue and was described as next-generation IHC. Robust, accurate, and high-throughput cell segmentation is crucial for the analysis of this new generation of IHC data. To this end, we propose a watershed-based cell segmentation, which uses a nuclear marker and multiple membrane markers, the latter automatically selected based on their correlation. In comparison with the state-of-the-art segmentation pipelines, which are only using a single marker for object detection, we could show that the use of multiple markers can significantly increase the segmentation power, and thus, multiplexed information should be used and not ignored during the segmentation. Furthermore, we provide a novel, user-friendly open-source toolbox for the automatic segmentation of multiplexed histopathological images.

Published
2014

6. Systematic analysis of cell phenotypes and cellular social networks in tissues using the histology topography cytometry analysis toolbox (histoCAT)

Author

Schapiro, Denis, primary, Jackson, Hartland Warren, additional, Raghuraman, Swetha, additional, Zanotelli, Vito R T, additional, Fischer, Jana R, additional, Schulz, Daniel, additional, Giesen, Charlotte, additional, Catena, Raúl, additional, Varga, Zsuzsanna, additional, and Bodenmiller, Bernd, additional

Published
2017
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9. Highly multiplexed imaging of tumor tissues with subcellular resolution by mass cytometry

Author

Giesen, Charlotte, primary, Wang, Hao A O, additional, Schapiro, Denis, additional, Zivanovic, Nevena, additional, Jacobs, Andrea, additional, Hattendorf, Bodo, additional, Schüffler, Peter J, additional, Grolimund, Daniel, additional, Buhmann, Joachim M, additional, Brandt, Simone, additional, Varga, Zsuzsanna, additional, Wild, Peter J, additional, Günther, Detlef, additional, and Bodenmiller, Bernd, additional

Published
2014
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10. LA-ICP-MS and nHPLC-ESI-LTQ-FT-MS/MS for the analysis of cisplatin–protein complexes separated by two dimensional gel electrophoresis in biological samples

Author

Moreno Gordaliza, Estefanía, Esteban Fernández, Diego, Giesen, Charlotte, Lehmann, Karola, Lázaro Fernández, Alberto, Tejedor, Alberto, Scheler, Christian, Cañas Montalvo, Benito, Jakubowski, Norbert, Linscheid, Michael W., Gómez Gómez, María Milagros, Moreno Gordaliza, Estefanía, Esteban Fernández, Diego, Giesen, Charlotte, Lehmann, Karola, Lázaro Fernández, Alberto, Tejedor, Alberto, Scheler, Christian, Cañas Montalvo, Benito, Jakubowski, Norbert, Linscheid, Michael W., and Gómez Gómez, María Milagros

Abstract

A method for the analysis of Pt–protein complexes in biological samples, previously subjected to cisplatin treatment, has been developed. Proteins were separated by gel electrophoresis, and those bound to Pt were detected with high sensitivity by LA-ICP-(SF)-MS. Pt-containing spots were in-gel digested with trypsin, and the peptides produced identified using nHPLC-ESI-LTQ-FT-MS/MS. The influence of protein separation conditions, staining and gel processing prior to laser ablation on Pt–protein bonds preservation have been evaluated using standard proteins incubated with cisplatin. 2-DE separation under non-reducing conditions followed by either Coomassie blue brilliant or silver staining is appropriate for Pt–protein complexes, achieving a good separating resolution of the proteins in biological samples. Direct LA-ICP-MS analysis of glycerol-treated dried gels for Pt–protein monitoring resulted in better sensitivity, more reliable relative Pt signals and a simpler and less timeconsuming approach compared to the analysis of blotted membranes. Ablation of gels allowed tackling protein identification of Pt-spots in the remaining non-ablated material in the gel, making it unnecessary to run several gels in parallel for separate Pt detection and protein identification. By using this approach, Pt coordinated to proteins, such as a-2-macroglobulin, transferrin, albumin or hemoglobin, was detected in the serum from a rat treated in vivo with cisplatin after nrSDS-PAGE separation. Furthermore, the first complete LA-ICP-MS metalloprotein contour map in a 2-DE gel has been produced, in this case for the detection of Pt–protein complexes in renal proximal tubule epithelial cells (RPTECs) incubated with cisplatin. Several proteins were identified in those spots containing Pt, which may have a connection with the drug-induced nephrotoxicity mainly affecting this cell type in the kidney., CICYT Projects, Depto. de Química Analítica, Fac. de Ciencias Químicas, TRUE, pub

Published
2012

14. LA-ICP-MS and nHPLC-ESI-LTQ-FT-MS/MS for the analysis of cisplatin–protein complexes separated by two dimensional gel electrophoresis in biological samples

Author

Moreno-Gordaliza, Estefanía, primary, Esteban-Fernández, Diego, additional, Giesen, Charlotte, additional, Lehmann, Karola, additional, Lázaro, Alberto, additional, Tejedor, Alberto, additional, Scheler, Christian, additional, Cañas, Benito, additional, Jakubowski, Norbert, additional, Linscheid, Michael W., additional, and Gómez-Gómez, M. Milagros, additional

Published
2012
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16. Elemental Bioimaging in Kidney by LA–ICP–MS As a Tool to Study Nephrotoxicity and Renal Protective Strategies in Cisplatin Therapies

Author

Moreno-Gordaliza, Estefanía, primary, Giesen, Charlotte, additional, Lázaro, Alberto, additional, Esteban-Fernández, Diego, additional, Humanes, Blanca, additional, Cañas, Benito, additional, Panne, Ulrich, additional, Tejedor, Alberto, additional, Jakubowski, Norbert, additional, and Gómez-Gómez, M. Milagros, additional

Published
2011
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