Journal
ANALYTICAL CHEMISTRY
Volume 82, Issue 2, Pages 491-497Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac902139w
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Funding
- NIH [P50-GM073197, U54 GM074961, P50-GM088499]
- NSF [MRI 0722558, CHE 0640549]
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Second-order nonlinear optical imaging of chiral crystals (SONICC) is explored for selective detection of integral membrane protein crystals grown in opaque and turbid environments. High turbidity is a hallmark of membrane protein crystallization due to the extensive use of detergent and/or lipids that often form various mesophases. Detection of crystals in such media by conventional optical methods (e.g., intrinsic LTV fluorescence, birefringence, bright-field image analysis, etc.) is often complicated by optical scattering and by the small sizes of the crystals that routinely form. SONICC is shown to be well-suited for this application, by nature of its compatibility with imaging in scattering media and its high selectivity protein crystals. Bright second harmonic generatioiri (SHG) (up to 18 million counts/s) was observed from; even relatively small crystals (5 pin) with a minimal background due to the surrounding lipid mesophase (similar to 1 thousand counts/s). The low background nature of the resulting protein crystal images permitted the use of a relatively simple, particle counting analysis for preliminary scoring. Comparisons between a particle counting analysis of SONICC images and protocols based on the human expert analysis of conventional bright-field and birefringence images were performed.
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