Nonlinear Optical Characterization of Membrane Protein Microcrystals and Nanocrystals
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.
KeywordsNonlinear optical microscopy Second harmonic generation Two-photon excited UV fluorescence Microscopy High-throughput screening Crystal detection Crystal positioning Crystal optimisation
The authors wish to acknowledge support from the National Institutes of Health (NIH) grants NIH-R01GM103401 and NIH-R01GM103910. We would also like to thank Ellen Gualtieri from Formulatrix for supplying the glucose isomerase crystal images in Fig. 7.2.
- Boyd RW (2008) Nonlinear optics, 3rd edn. Elsevier Inc, CambridgeGoogle Scholar
- Madden JT, Toth SJ, Dettmar CM, Newman JA, Oglesbee RA, Hedderich HG, Everly RM, Becker M, Ronau JA, Buchanan SK et al (2013) Integrated nonlinear optical imaging microscope for on-axis crystal detection and centering at a synchrotron beamline. J Synchrotron Radiat 20(4):531–540CrossRefGoogle Scholar