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Cell Signal. 2019 Oct;62:109335. doi: 10.1016/j.cellsig.2019.05.020. Epub 2019 Jun 3.

New automatic quantification method of immunofluorescence and histochemistry in whole histological sections.

Author information

1
Experimental Nephrology and Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: Friederike.Kessel@uniklinikum-dresden.de.
2
Experimental Nephrology and Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: Anne.Steglich2@uniklinikum-dresden.de.
3
Experimental Nephrology and Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: tschoto@outlook.de.
4
Experimental Nephrology and Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: Florian.Gembardt@uniklinikum-dresden.de.
5
Experimental Nephrology and Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: Leo.Ruhnke@uniklinikum-dresden.de.
6
Experimental Nephrology and Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: Julian.Stumpf@uniklinikum-dresden.de.
7
Institute for Immunology, Carl Gustav Carus Faculty of Medicine, Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: rayk.behrendt@tu-dresden.de.
8
Paul Langerhans Institute Dresden (PLID) of Helmholtz Zentrum München at the University Hospital Carl Gustav Carus at the Technische Universität Dresden, Tatzberg 47, 01307 Dresden, Helmholtz Zentrum München, München-Neuherberg, Germany.; German Center for Diabetes Research (DZD), Ingolstädter Landstrasse 1, 85764 München-Neuherberg, Germany. Electronic address: Christian.Cohrs@tu-dresden.de.
9
Department of Physiology, Carl Gustav Carus Faculty of Medicine, Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: irakli.kopaliani@tu-dresden.de.
10
Experimental Nephrology and Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: Vladimir.Todorov@uniklinikum-dresden.de.
11
Experimental Nephrology and Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: Michael.Gerlach@uniklinikum-dresden.de.
12
Experimental Nephrology and Division of Nephrology, Department of Internal Medicine III, University Hospital Carl Gustav Carus at the Technische Universität Dresden, Fetscherstrasse 74, 01307 Dresden, Germany. Electronic address: Christian.Hugo@uniklinikum-dresden.de.

Abstract

Immunofluorescent staining is a widespread tool in basic science to understand organ morphology and (patho-) physiology. The analysis of imaging data is often performed manually, limiting throughput and introducing human bias. Quantitative analysis is particularly challenging for organs with complex structure such as the kidney. In this study we present an approach for automatic quantification of fluorescent markers and histochemical stainings in whole organ sections using open source software. We validate our novel method in multiple typical challenges of basic kidney research and demonstrate its general relevance and applicability to other complex solid organs for a variety of different markers and stainings. Our newly developed software tool "AQUISTO", applied as a standard in primary data analysis, facilitates efficient large scale evaluation of cellular populations in various types of histological samples. Thereby it contributes to the characterization and understanding of (patho-) physiological processes.

KEYWORDS:

Automatic quantification; Cell count; Histochemistry; Immunofluorescence; Morphology

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