New techniques for detecting and handling nanocrystals for cutting edge structural biology methods

Project Summary Determining the detailed structural characteristics of biomolecules relevant to human health and disease is one of the most crucial tools in our arsenal for understanding disease etiology and mechanism, and for being able to develop new therapeutics that target these molecular entities. There are new techniques in structural biology, including serial femtosecond crystallography, serial synchrotron crystallography, and microcrystal electron diffraction, that have the potential to greatly advance structure determination of biomolecules and to empower access to structural details that have defied characterization via other structural methods. These new structural methods all rely on being able to generate, detect and appropriately handle extremely small crystalline samples of biomolecules. This requirement for sub- micron sized crystals is one of the key features of these technologies, and presents a major obstacle to the advancement of these methods for structure determination. This proposal presents innovative technologies for both image analysis and sample handling expressly designed to address the specific challenges of working with submicron crystals. We plan to use nonlinear optical microscopy methods coupled with purpose-built application of point process modeling and wavelet image analysis approaches to provide computational tools needed to enable detection and characterization of submicron samples that are invisible to the brightfield microscopy tools that are typically used in sample generation and experimental set up for crystal based structural biology. In addition, we will examine different fixed target platforms to reduce sample handling, minimizing potential crystal damage, as well as test use of acoustic droplet ejection techniques for nanoliter volume sample transfer. These innovations will be a powerful addition to structural biology toolbox for leveraging the cutting edge diffraction based methods currently available for structure determination. These technology developments will break through key barriers to the widespread use of these cutting edge structural methods.