Biophysics

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    Quantitative Analysis of Clot Deposition on Extracorporeal Life Support Membrane Oxygenators Using Digital and Scanning Electron Microscopy Imaging Techniques

    Device-induced thrombosis remains a major complication of extracorporeal life support (ECLS). To more thoroughly understand how blood components interact with the artificial surfaces of ECLS circuit components, assessment of clot deposition on these

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    Perforated Patch Clamp Recordings in ex vivo Brain Slices from Adult Mice
    Authors:  Simon Hess, Helmut Wratil and Peter Kloppenburg, date: 08/20/2023, view: 357, Q&A: 0

    Intracellular signaling pathways directly and indirectly regulate neuronal activity. In cellular electrophysiological measurements with sharp electrodes or whole-cell patch clamp recordings, there is a great risk that these signaling pathways are

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    Protein Structure Predictions, Atomic Model Building, and Validation Using a Cryo-EM Density Map from Hepatitis B Virus Spherical Subviral Particle
    Authors:  Nadia DiNunno, Emily N. Bianchini, Haitao Liu and Joseph Che-Yen Wang, date: 07/20/2023, view: 490, Q&A: 0

    Hepatitis B virus (HBV) infection is a global public health concern. During chronic infection, the HBV small-surface antigen is expressed in large excess as non-infectious spherical subviral particles (SVPs), which possess strong immunogenicity. To

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    An ex vivo Model of Paired Cultured Hippocampal Neurons for Bi-directionally Studying Synaptic Transmission and Plasticity
    Authors:  Ruslan Stanika and Gerald J. Obermair, date: 07/20/2023, view: 405, Q&A: 0

    Synapses provide the main route of signal transduction within neuronal networks. Many factors regulate critical synaptic functions. These include presynaptic calcium channels, triggering neurotransmitter release, and postsynaptic ionotropic

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    Recoil Measurements in Drosophila Embryos: from Mounting to Image Analysis
    Authors:  Luis Eduardo Sánchez-Cisneros, Sourabh Bhide and Luis Daniel Ríos-Barrera, date: 07/20/2023, view: 309, Q&A: 0

    Tension and force propagation play a central role in tissue morphogenesis, as they enable sub- and supra-cellular shape changes required for the generation of new structures. Force is often generated by the cytoskeleton, which forms complex

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    Biophysical Analysis of Mechanical Signals in Immotile Cilia of Mouse Embryonic Nodes Using Advanced Microscopic Techniques
    Authors:  Takanobu A. Katoh, Toshihiro Omori, Takuji Ishikawa, Yasushi Okada and Hiroshi Hamada, date: 07/20/2023, view: 604, Q&A: 0

    Immotile cilia of crown cells at the node of mouse embryos are required for sensing leftward fluid flow that gives rise to the breaking of left-right (L-R) symmetry. The flow-sensing mechanism has long remained elusive, mainly because of

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    Dual-color Colocalization in Single-molecule Localization Microscopy to Determine the Oligomeric State of Proteins in the Plasma Membrane

    Determining the oligomeric state of membrane proteins is critical for understanding their function. However, traditional ex situ methods like clear native gel electrophoresis can disrupt protein subunit interactions during sample preparation. In

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    Synthesis and Purification of Lipid-conjugated Fluorescent pH Sensors
    Authors:  Wiebke Wiesner, Ronja Marie Kühnel and Thomas Guenther Pomorski, date: 06/05/2023, view: 451, Q&A: 0

    Lipid-conjugated pH sensors based on fluorophores coupled to lipids are a powerful tool for monitoring pH gradients in biological microcompartments and reconstituted membrane systems. This protocol describes the synthesis of pH sensors based on

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    Measuring in vitro ATPase Activity with High Sensitivity Using Radiolabeled ATP
    Authors:  Sarina Veit and Thomas Günther Pomorski, date: 05/20/2023, view: 404, Q&A: 0

    ATPase assays are a common tool for the characterization of purified ATPases. Here, we describe a radioactive [γ-32P]-ATP-based approach, utilizing complex formation with molybdate for phase separation of the free phosphate from

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    In vitro Reconstitution of Phase-separated p62 Bodies on the Arp2/3-derived Actin Network
    Authors:  Tong Liu, Mengbo Xu and Na Mi, date: 04/20/2023, view: 413, Q&A: 0

    In cells, p62/SQSTM1 undergoes liquid–liquid phase separation (LLPS) with poly-ubiquitin chains to form p62 bodies that work as a hub for various cellular events, including selective autophagy. Cytoskeleton components such as Arp2/3-derived

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