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University of saarland

8 ARTICLES PUBLISHED IN JoVE

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Medicine

Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia
Yves Harder 1, Daniel Schmauss 1, Reto Wettstein 2, José T. Egaña 1, Fabian Weiss 1, Andrea Weinzierl 1, Anna Schuldt 1, Hans-Günther Machens 1, Michael D. Menger 3, Farid Rezaeian 4
1Department of Plastic Surgery and Hand Surgery, Klinikum rechts der Isar, Technische Universität München, 2Department of Plastic, Reconstructive, Aesthetic and Hand Surgery, University Hospital of Basel, 3Institute for Clinical and Experimental Surgery, University of Saarland, 4Division of Plastic and Hand Surgery, University Hospital Zurich

The window of the murine dorsal skinfold chamber presented visualizes a zone of acute persistent ischemia of a musculocutaneous flap. Intravital epi-fluorescence microscopy permits for direct and repetitive assessment of the microvasculature and quantification of hemodynamics. Morphologic and hemodynamic results can further be correlated with histological and molecular analyses.

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Bioengineering

Studying the Stoichiometry of Epidermal Growth Factor Receptor in Intact Cells using Correlative Microscopy
Diana B. Peckys 1, Niels de Jonge 1,2
1INM-Leibniz Institute for New Materials, 2Department of Physics, University of Saarland

This protocol describes the labeling of epidermal growth factor receptor (EGFR) on COS7 fibroblast cells, and subsequent correlative light- and electron microscopy of whole cells in hydrated state. The label contained fluorescent quantum dots. The protocol can be used to study the stoichiometry of EGFR at the single molecule level.

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Medicine

An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice
Tina Histing 1, Michael D. Menger 2, Tim Pohlemann 1, Romano Matthys 3, Tobias Fritz 1, Patric Garcia 1, Moritz Klein 1
1Department of Trauma, Hand and Reconstructive Surgery, Saarland University, 2Institute for Clinical & Experimental Surgery, Saarland University, 3RISystem AG

This protocol describes an osteosynthesis technique using an intramedullary locking nail for standardized fixation of femur osteotomies, which can be used to analyze normal and defective bone healing in mice.

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Engineering

Studying Dynamic Processes of Nano-sized Objects in Liquid using Scanning Transmission Electron Microscopy
Justus Hermannsdörfer 1, Niels de Jonge 1,2
1INM-Leibniz Institute for New Materials, 2Department of Physics, University of Saarland

This protocol describes the operation of a liquid flow specimen holder for scanning transmission electron microscopy of AuNPs in water, as used for the observation of nanoscale dynamic processes.

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Developmental Biology

Isolation and Enrichment of Liver Progenitor Subsets Identified by a Novel Surface Marker Combination
Henrike Julich-Haertel 1, Marina Tiwari 1, Christina Mehrfeld 1, Elmar Krause 2, Miroslaw Kornek 1, Veronika Lukacs-Kornek 1
1Department of Medicine II, Saarland University Medical Center, 2Department of Physiology, Center for Integrative Physiology and Molecular Medicine (CIPMM), University of Saarland

Liver injuries are accompanied by progenitor cell expansion that represents a heterogeneous cell population. Novel classification of this cellular compartment allows for the distinguishing of multiple subsets. The method described here illustrates the flow cytometry analysis and high purity isolation of various subsets that can be used for further assays.

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Bioengineering

Isolation of Murine Adipose Tissue-derived Microvascular Fragments as Vascularization Units for Tissue Engineering
Florian S. Frueh 1,2, Thomas Später 1, Claudia Scheuer 1, Michael D. Menger 1, Matthias W. Laschke 1
1Institute for Clinical and Experimental Surgery, Saarland University, 2Division of Plastic Surgery and Hand Surgery, University Hospital Zurich, University of Zurich

We present a protocol to isolate adipose tissue-derived microvascular fragments that represent promising vascularization units. They can be rapidly isolated, do not require in vitro processing and, thus, may be used for one-step prevascularization in different fields of tissue engineering.

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Medicine

A Minimally Invasive Model to Analyze Endochondral Fracture Healing in Mice Under Standardized Biomechanical Conditions
Tina Histing 1, Philipp Bremer 1, Mika F. Rollmann 1, Steven Herath 1, Moritz Klein 1, Tim Pohlemann 1, Michael D. Menger 2, Tobias Fritz 1
1Department of Trauma, Hand and Reconstructive Surgery, Saarland University, 2Institute for Clinical & Experimental Surgery, Saarland University

This protocol describes a minimally invasive osteosynthesis technique using an intramedullary screw for standardized stabilization of femur fractures, which can be used to analyze endochondral bone healing in mice.

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Biology

Graphene Enclosure of Chemically Fixed Mammalian Cells for Liquid-Phase Electron Microscopy
Patricia Blach 1,2, Sercan Keskin 1, Niels de Jonge 1,2
1INM-Leibniz Institute for New Materials, 2Department of Physics, Saarland University

Presented here is a protocol for labeling membrane proteins in mammalian cells and coating the sample with graphene for liquid-phase scanning transmission electron microscopy. The stability of the samples against the damage caused by radiation can also studied with this protocol.

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