Selective Detection of Protein Crystals by Second Harmonic Microscopy
Citations Over TimeTop 10% of 2008 papers
Abstract
The unique symmetry properties of second harmonic generation (SHG) microscopy enabled sensitive and selective imaging of protein microcrystals with negligible contributions from solvated proteins or amorphous protein aggregates. In studies of microcrystallites of green fluorescent protein (GFP) prepared in 500 pL droplets, the SHG intensities rivaled those of fluorescence, but with superb selectivity for crystalline regions. GFP in amorphous aggregates and in solution produced substantial background fluorescence, but no detectable SHG. The ratio of the forward-to-backward detected SHG provides a measure of the particle size, suggesting detection limits down to crystallites 100 nm in diameter under low magnification (10x). In addition to being sensitive and highly selective, second-order nonlinear optical imaging of chiral crystals (SONICC) is directly compatibility with virtually all common protein crystallization platforms.
Related Papers
- → Second-Harmonic Generation Imaging Microscopy of Structural Protein Arrays in Tissue(1998)1 cited
- Quantitative Analysis of Backward Generation and Backscattering for Epi-collected Second Harmonic Generation in Biological Tissues(2007)
- Development of Cell Image Based on Second Harmonic Generation(2008)
- Microscopic Imaging Technology of Second Harmonic Generation(2008)
- → Circular Dichroism Second-Harmonic Generation Imaging of KTiOPO4 Nanocrystal Through Stratified Media(2022)