Nonlinear Optical Characterization of Membrane Protein Microcrystals and Nanocrystals

Adv Exp Med Biol. 2016;922:91-103. doi: 10.1007/978-3-319-35072-1_7.


Nonlinear optical methods such as second harmonic generation (SHG) and two-photon excited UV fluorescence (TPE-UVF) imaging are promising approaches to address bottlenecks in the membrane protein structure determination pipeline. The general principles of SHG and TPE-UVF are discussed here along with instrument design considerations. Comparisons to conventional methods in high throughput crystallization condition screening and crystal quality assessment prior to X-ray diffraction are also discussed.

Keywords: Crystal detection; Crystal optimisation; Crystal positioning; High-throughput screening; Microscopy; Nonlinear optical microscopy; Second harmonic generation; Two-photon excited UV fluorescence.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Crystallization
  • Crystallography, X-Ray
  • Equipment Design
  • High-Throughput Screening Assays
  • Humans
  • Lasers
  • Membrane Proteins / chemistry*
  • Microscopy / instrumentation
  • Microscopy / methods*
  • Microscopy, Fluorescence, Multiphoton / instrumentation
  • Microscopy, Fluorescence, Multiphoton / methods
  • Microscopy, Polarization / instrumentation
  • Microscopy, Polarization / methods
  • Nanoparticles / chemistry*
  • Scattering, Radiation
  • Time Factors
  • Ultraviolet Rays


  • Membrane Proteins