Muscle diffraction at the life science x-ray scattering beamline

We report on recent methodological advances at the Life Science X-ray Scattering (LiX) beamline of the National Synchrotron Light Source II (NSLS-II) to support small-angle X-ray scattering experiments on skeletal and cardiac muscle tissues. These experiments have been routinely performed at the BioCAT beamline of the Advanced Photon Source (APS) over the past two decades to measure sarcomeric protein organization within healthy and diseased muscle tissues and provide direct molecular evidence for their functional roles and dynamics. Many recent advances in our understanding of sarcomeric proteins relied on diffraction data and include, as examples, MyBP-C, crossbridge SRX/DRX states, and titin. With LiX now available for muscle experimentation, more muscle users can be supported which will speed up research of sarcomeric proteins, muscle biomechanics, and skeletal and cardiac myopathies. LiX explicitly focuses on high-throughput muscle diffraction with rapid sample turnover and semi-automated data processing. These operations have been tested and validated on skeletal and cardiac tissues sourced from both humans and multiple animal models including pig, rat, mouse, and zebrafish.

Methods
Processed X-ray diffraction data of mouse skeletal EDL muscles at 2.7um sarcomere length and room temperature (~22C) under three states: Control, + tobacco etch virus protease, and + mavacamten. Data was acquired at the LiX beamline of NSLS-II. The mice were SnoopC2, an engineered line that controllably cleaves away N-terminal fast myosin binding protein C upon incubation with tobacco etch virus protease.

Data were saved as .tiff files with floating point precision. An image of AgBH is also provided as a calibration standard.