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Ludwig Maximilian University of Munich

5 ARTICLES PUBLISHED IN JoVE

JoVE Core

20 mJ, 1 ps Yb:YAG Thin-disk Regenerative Amplifier

Ayman Alismail^1,2, Haochuan Wang^1,3, Jonathan Brons³, Hanieh Fattahi^1,3

¹Department of Physics, Ludwig Maximilian University of Munich, ²Physics and Astronomy Department, King Saud University, ³Max Planck Institute of Quantum Optics

A protocol for the operation of a high-energy, high-power optical parametric chirped pulse amplifier pump source based on an Yb:YAG thin-disk regenerative amplifier is presented here.

Genetics

A Customizable Protocol for String Assembly gRNA Cloning (STAgR)

Christopher T. Breunig^1,2, Andrea M. Neuner^1,2, Jessica Giehrl-Schwab³, Wolfgang Wurst³, Magdalena Götz^2,4, Stefan H. Stricker^1,2

¹MCN Junior Research Group, Munich Center for Neurosciences, Ludwig Maximilian Universitat, BioMedical Center, ²Institute of Stem Cell Research, Helmholtz Zentrum, German Research Center for Environmental Health, ³Institute of Developmental Genetics, Helmholtz Zentrum, German Research Center for Environmental Health, ⁴Physiological Genomics, Ludwig Maximilian Universitat, BioMedical Center

Here, we present string assembly gRNA cloning (STAgR), a method to easily multiplex gRNA vectors for CRISPR/Cas9 approaches. STAgR makes gRNA multiplexing simple, efficient and customizable.

Medicine

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

Martin Ryll¹, Julian Bucher¹, Moritz Drefs¹, Florian Bösch¹, K. Kumaraswami², Tobias Schiergens¹, Hanno Niess¹, Markus Schoenberg¹, Sven Jacob¹, Markus Rentsch¹, Markus Guba¹, Jens Werner¹, Joachim Andrassy¹, Michael N. Thomas^1,3

¹Department of General, Visceral, and Transplant Surgery, Ludwig Maximilian University of Munich, ²Walter-Brendel-Centre of Experimental Medicine, Ludwig Maximilian University of Munich, ³Department of General, Visceral and Cancer Surgery, University Hospital of Cologne, University of Cologne

Here, we present a protocol of heterotopic aortic transplantation in mice using the non-suture cuff technique in a cervical murine model. This model can be used to study the underlying pathology of chronic allograft vasculopathy (CAV) and can help evaluate new therapeutic agents in order to prevent its formation.

Neuroscience

Cryo-section Dissection of the Adult Subependymal Zone for Accurate and Deep Quantitative Proteome Analysis

Christian Friess¹, Magdalena Götz^1,2,3, Jacob Kjell^1,2,4

¹Division of Physiological Genomics, Biomedical Center, Ludwig Maximilian University of Munich, ²Institute for Stem Cell Research, Helmholtz Zentrum München, ³SYNERGY, Excellence Cluster Systems Neurology, University of Munich, ⁴Department of Clinical Neuroscience, Karolinska Institutet

Cryo-section-dissection allows fresh, frozen preparation of the largest neurogenic niche in the murine brain for deep quantitative proteome analysis. The method is precise, efficient, and causes minimal tissue perturbation. Therefore, it is ideally suited for studying the molecular microenvironment of this niche, as well as other organs, regions, and species.

Biology

Generation and Maintenance of Primate Induced Pluripotent Stem Cells Derived from Urine

Jessica Radmer¹, Johanna Geuder¹, Fiona C. Edenhofer¹, Wolfgang Enard¹, Mari Ohnuki^1,2,3

¹Faculty of Biology, Ludwig Maximilian University of Munich, ²Institute for the Advanced Study of Human Biology, Kyoto University, ³Hakubi Center, Kyoto University

The present protocol describes a method to isolate, expand, and reprogram human and non-human primate urine-derived cells to induced pluripotent stem cells (iPSCs), as well as instructions for feeder-free maintenance of the newly generated iPSCs.