Protein-crystal detection with a compact multimodal multiphoton microscope

Commun Biol. 2020 Oct 13;3(1):569. doi: 10.1038/s42003-020-01275-8.

Abstract

There is an increasing demand for rapid, effective methods to identify and detect protein micro- and nano-crystal suspensions for serial diffraction data collection at X-ray free-electron lasers or high-intensity micro-focus synchrotron radiation sources. Here, we demonstrate a compact multimodal, multiphoton microscope, driven by a fiber-based ultrafast laser, enabling excitation wavelengths at 775 nm and 1300 nm for nonlinear optical imaging, which simultaneously records second-harmonic generation, third-harmonic generation and three-photon excited ultraviolet fluorescence to identify and detect protein crystals with high sensitivity. The instrument serves as a valuable and important tool supporting sample scoring and sample optimization in biomolecular crystallography, which we hope will increase the capabilities and productivity of serial diffraction data collection in the future.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallization / methods
  • Lab-On-A-Chip Devices
  • Liquid Crystals*
  • Microscopy, Fluorescence, Multiphoton* / methods
  • Models, Molecular*
  • Protein Conformation
  • Proteins / chemistry*
  • Reproducibility of Results
  • Structure-Activity Relationship

Substances

  • Proteins