Max Planck Institute of Biochemistry

7 articles published in JoVE

• 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.

• Bioengineering

  
  Rapid Encapsulation of Reconstituted Cytoskeleton Inside Giant Unilamellar Vesicles Yashar Bashirzadeh*¹, Nadab Wubshet*¹, Thomas Litschel², Petra Schwille³, Allen P. Liu^{1,4,5,6 1}Department of Mechanical Engineering, University of Michigan, Ann Arbor, ²John A. Paulson School of Engineering and Applied Sciences, Harvard University, ³Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, ⁴Department of Biomedical Engineering, University of Michigan, Ann Arbor, ⁵Department of Biophysics, University of Michigan, Ann Arbor, ⁶Cellular and Molecular Biology Program, University of Michigan, Ann Arbor This article introduces a simple method for expeditious production of giant unilamellar vesicles with encapsulated cytoskeletal proteins. The method proves to be useful for bottom-up reconstitution of cytoskeletal structures in confinement and cytoskeleton-membrane interactions.

• Biochemistry

  
  In Vitro Reconstitution of Self-Organizing Protein Patterns on Supported Lipid Bilayers Beatrice Ramm*¹, Philipp Glock*¹, Petra Schwille^{1 1}Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry We provide a protocol for in vitro self-organization assays of MinD and MinE on a supported lipid bilayer in an open chamber. Additionally, we describe how to enclose the assay in lipid-clad PDMS microcompartments to mimic in vivo conditions by reaction confinement.

• Chemistry

  
  Microfluidic Chips for In Situ Crystal X-ray Diffraction and In Situ Dynamic Light Scattering for Serial Crystallography Yannig Gicquel*^1,2, Robin Schubert*^3,4,5, Svetlana Kapis³, Gleb Bourenkov⁶, Thomas Schneider⁶, Markus Perbandt^3,4, Christian Betzel^3,4,5, Henry N. Chapman^1,2,4, Michael Heymann^{1,7 1}Center for Free Electron Laser Science, DESY, ²Department of Physics, University of Hamburg, ³Institute for Biochemistry and Molecular Biology, Laboratory for Structural Biology of Infection and Inflammation, University of Hamburg, ⁴The Hamburg Center for Ultrafast Imaging, University of Hamburg, ⁵Integrated Biology Infrastructure Life-Science Facility at the European XFEL (XBI), ⁶European Molecular Biology Laboratory, EMBL c/o DESY, ⁷Department of Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry This protocol describes in detail how to fabricate and operate microfluidic devices for X-ray diffraction data collection at room temperature. Additionally, it describes how to monitor protein crystallization by dynamic light scattering and how to process and analyze obtained diffraction data.

• Developmental Biology

  
  In Vivo Imaging of Muscle-tendon Morphogenesis in Drosophila Pupae Sandra B. Lemke¹, Frank Schnorrer^{1,2 1}Muscle Dynamics Group, Max Planck Institute of Biochemistry, ²Aix Marseille University, CNRS, IBDM Here, we present an easy-to-use and versatile method to perform live imaging of developmental processes in general and muscle-tendon morphogenesis in particular in living Drosophila pupae.

• Biology

  
  Proteomic Sample Preparation from Formalin Fixed and Paraffin Embedded Tissue Jacek R. Wiśniewski^{1 1}Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry Archival formalin fixed and paraffin embedded (FFPE) clinical samples are valuable material for investigation of diseases. Here we demonstrate a sample preparation workflow allowing in-depth proteomic analysis of microdissected FFPE tissue.

• Bioengineering

  
  Visualization of Cortex Organization and Dynamics in Microorganisms, using Total Internal Reflection Fluorescence Microscopy Felix Spira¹, Julia Dominguez-Escobar¹, Nikola Müller^1,2, Roland Wedlich-Söldner^{1 1}AG Cellular Dynamics and Cell Patterning, Max Planck Institute of Biochemistry, ²Helmholtz Zentrum München Total Internal Reflection Fluorescence (TIRF) microscopy is a powerful approach to observe structures close to the cell surface at high contrast and temporal resolution. We demonstrate how TIRF can be employed to study protein dynamics at the cortex of cell wall-enclosed bacterial and fungal cells.