Technical University Munich

4 articles published in JoVE

• Biology

  
  Isolation of Nuclei from Flash-frozen Liver Tissue for Single-cell Multiomics Mateusz Strzelecki^1,4, Kelvin Yin¹, Carlos Talavera-López^2,3, Celia P. Martinez-Jimenez^{1,4 1}Helmholtz Pioneer Campus (HPC), Helmholtz Munich, ²Institute of Computational Biology, Computational Health Department, Helmholtz Munich, ³Division of Infectious Diseases and Tropical Medicine, Ludwig-Maximillian-Universität Klinikum, ⁴TUM School of Medicine, Technical University of Munich Here, we present a protocol for isolating nuclei from flash-frozen, archived liver tissues for single-nucleus RNA-seq, ATAC-seq, and joint multiomics (RNA-seq and ATAC-seq).

• Neuroscience

  
  Microfluidics-Assisted Selective Depolarization of Axonal Mitochondria Simone Wanderoy*^1,2, Alina Rühmkorf*^1,2, Angelika B. Harbauer^{2,3,4 1}TUM Medical Graduate Center, Technical University of Munich, ²Max Planck Institute of Neurobiology, ³TUM School of Medicine, Institute of Neuronal Cell Biology, Technical University of Munich, ⁴Munich Cluster for Systems Neurology The present protocol describes the seeding and staining of neuronal mitochondria in microfluidic chambers. The fluidic pressure gradient in these chambers allows for the selective treatment of mitochondria in axons to analyze their properties in response to pharmacological challenges without affecting the cell body compartment.

• Medicine

  
  Combining 3D-Printing and Electrospinning to Manufacture Biomimetic Heart Valve Leaflets Benedikt Freystetter¹, Maximilian Grab^1,2, Linda Grefen¹, Lara Bischof¹, Lorenz Isert³, Petra Mela², Deon Bezuidenhout⁴, Christian Hagl^1,5, Nikolaus Thierfelder^{1 1}Department of Cardiac Surgery, Ludwig Maximilians University Munich, ²Chair of Medical Materials and Implants, Technical University Munich, ³Faculty for Chemistry and Pharmacy, Ludwig Maximilians University Munich, ⁴Cardiovascular Research Unit, University of Cape Town, ⁵DZHK (German Centre for Cardiovascular Research), partner site Munich Heart Alliance The presented method offers an innovative way for engineering biomimetic fiber structures in three-dimensional (3D) scaffolds (e.g., heart valve leaflets). 3D-printed, conductive geometries were used to determine shape and dimensions. Fiber orientation and characteristics were individually adjustable for each layer. Multiple samples could be manufactured in one setup.

• Biochemistry

  
  Mass-Sensitive Particle Tracking to Characterize Membrane-Associated Macromolecule Dynamics Frederik Steiert^1,2, Tamara Heermann¹, Nikolas Hundt³, Petra Schwille^{1 1}Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, ²Department of Physics, Technical University Munich, ³Department of Cellular Physiology, Biomedical Center (BMC), Ludwig-Maximilians-Universität München This protocol describes an iSCAT-based image processing and single-particle tracking approach that enables the simultaneous investigation of the molecular mass and the diffusive behavior of macromolecules interacting with lipid membranes. Step-by-step instructions for sample preparation, mass-to-contrast conversion, movie acquisition, and post-processing are provided alongside directions to prevent potential pitfalls.