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Heinrich-Heine-University

6 ARTICLES PUBLISHED IN JoVE

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Biology

An Innovative Method for Exosome Quantification and Size Measurement
Arash Mehdiani 1, Anatol Maier 1, Antonio Pinto 1, Mareike Barth 1, Payam Akhyari 1, Artur Lichtenberg 1
1Research Group Experimental Surgery, Department of Cardiovascular Surgery, Medical Faculty, Heinrich-Heine-University Dusseldorf

A method for isolation of exosomes from whole blood and further analysis by nanoparticle tracking using a semi-automatic instrument is presented in this article. The presented technology provides an extremely sensitive method for visualizing and analyzing particles in liquid suspension.

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Biochemistry

Isolation of Glomeruli and In Vivo Labeling of Glomerular Cell Surface Proteins
Eva Königshausen 1, Sebastian A. Potthoff 1, Raphael Haase 1, Catherine Meyer-Schwesinger 2, Ernest Kaufmann 1, L. Christian Rump 1, Johannes Stegbauer 1, Lorenz Sellin 1, Ivo Quack 1, Magdalena Woznowski 1
1Department of Nephrology, Medical Faculty, Heinrich-Heine-University, 2Institute of Cellular and Integrative Physiology, University Medical Center Hamburg-Eppendorf

Here we present a protocol for murine in vivo labeling of glomerular cell surface proteins with biotin. This protocol contains information on how to perfuse mouse kidneys, isolate glomeruli, and perform endogenous immunoprecipitation of the protein of interest.

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Biochemistry

Rapid Fluorescence-based Characterization of Single Extracellular Vesicles in Human Blood with Nanoparticle-tracking Analysis
Andreas Weber 1, Julia Christin Wehmeyer 1, Vera Schmidt 1, Artur Lichtenberg 1, Payam Akhyari 1
1Department of Cardiovascular Surgery, Medical Faculty, Heinrich-Heine-University

In this protocol, we describe the complete workflow for rapid isolation of extracellular vesicles from human whole blood and characterization of specific markers by fluorescence-based nanoparticle-tracking analysis. The presented results show a high level of reproducibility and can be adjusted to cell culture supernatants.

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Medicine

Highly Sensitive Measurement of Glomerular Permeability in Mice with Fluorescein Isothiocyanate-polysucrose 70
Eva Königshausen 1, Sebastian A. Potthoff 1, Magdalena Woznowski 1, Johannes Stegbauer 1, Lars C. Rump 1, Lorenz Sellin 1
1Department of Nephrology, Medical Faculty, Heinrich-Heine-University

Here, we present a protocol to test glomerular permeability in mice using a highly sensitive, nonradioactive tracer. This method allows repetitive urine analyses with small urine volumes.

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Biochemistry

Imaging of Podocytic Proteins Nephrin, Actin, and Podocin with Expansion Microscopy
Eva Königshausen *1, Christina Theresa Schmitz *1, Lars Christian Rump 1, Lorenz Sellin 1
1Department of Nephrology, Medical Faculty, Heinrich-Heine-University

The presented method enables visualization of fluorescently labeled cellular proteins with expansion microscopy leading to a resolution of 70 nm on a conventional microscope.

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Genetics

Application of DNA Fingerprinting using the D1S80 Locus in Lab Classes
Meike Siebers *1, Agatha Walla *1,2, Thea Rütjes 1, Moritz-Fabian Müller 4, Maria von Korff 1,2,3
1Institute of Plant Genetics, Heinrich-Heine-University, 2Cluster of Excellence on Plant Sciences "SMART Plants for Tomorrow's Needs", 3Max Planck Institute for Plant Breeding Research, 4IBG-1: Biotechnology, Forschungszentrum Jülich GmbH

Here we describe a simple protocol to generate DNA fingerprinting profiles by amplifying the VNTR locus D1S80 from epithelial cell DNA.

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