Nonlinear optical methods for the analysis of protein nanocrystals and biological tissues

Ximeng You Dow, Purdue University

Abstract

Structural biology underpins rational drug design and fundamental understanding of protein function. X-ray diffraction (XRD) has been the golden standard for solving for high-resolution protein structure. Second harmonic generation (SHG) microscopy has been developed by the Simpson lab as a sensitive, crystal-specific detection method for the identification of protein crystal and help optimize the crystallization condition. Protein nanocrystals has been widely used for structure determination of membrane proteins in serial femtosecond nanocrystallography. In this thesis work, novel nonlinear optical methods were developed to address the challenges associated with the detection and characterization of protein nanocrystals. SHG-correlation spectroscopy (SHG-CS) was developed to take advantage of the diffusing motion and retrieve the size distribution and crystal quality of the nanocrystals. Polarization-dependent SHG imaging technique was developed to measure the relative orientation as well as the internal structure of the sample. Two photon- excited fluorescence has been used in the Simpson lab as a complementary measurement besides the inherent SHG signal from the crystals. A novel instrumentation development was also introduced in this thesis work to greatly improve the speed of fluorescence lifetime imaging (FLIM).

Degree

Ph.D.

Advisors

Simpson, Purdue University.

Subject Area

Analytical chemistry|Optics

Off-Campus Purdue Users:
To access this dissertation, please log in to our
proxy server
.

Share

COinS