Heterogeneity pervades biology. Among cells and subcellular structures like organelles there is a great deal of variability in terms of their size, chemical composition, and physiological properties. To understand these heterogeneous properties, researchers often study large groups of single cells, single organelles, or even single molecules. But, studying large samples of single entities comes with a cost. These measurements are usually low-throughput, meaning that many slow and labor-intensive individual measurements are required to gain new insights. To overcome this challenge and increase throughput, many groups have devised methods to rapidly analyze single cells, organelles, or other biological particles one-after-another. Alternatively, many individual objects can be analyzed simultaneously by arranging them into closely-packed arrays on a surface. The Wittenberg Lab is taking the latter approach for the high-throughput analysis of small vesicles produced by bacteria and isolated nerve terminals, called synaptosomes, acquired from brain tissue. To support this research, Prof. Nate Wittenberg recently received a grant from the National Institutes of Health for a project entitled, “Microfluidic Nanoarrays for High-throughput Analysis of Biological Nanostructures.” The grant will fund two graduate research assistantships through 2021. The overall goal of this project is to develop technology that enables the simultaneous analysis of hundreds to thousands of individual liposomes, bacterial vesicles, or synaptosomes. These particles are pattened by first creating a nanoscale array of capture spots on glass surfaces using a process that works like a small-scale rubber stamp. The capture spots are small enough (200 nanometer diameter) to capture just a single particle, packed closely together (approx. 1 micron spacing), and they are composed of molecules that specifically capture only the particles of interest. After patterning the capture spots, a microfluidic channel is placed over surface for delivery of particles and reagents to the array. In collaboration with Prof. Angela Brown (Lehigh Dept. of Chemical & Biomolecular Engineering), graduate students in the Wittenberg group will analyze large numbers of individual bacterial vesicles with fluorescence microscopy to determine how toxins are sorted to vesicles as a function of their size. The synaptosome arrays, also analyzed with fluorescence microscopy, will be used to measure heterogeneity in ion transport, membrane recycling, and neurotransmitter uptake and release. Nate Wittenberg was born and raised in Eau Claire, WI. He attended the University of Minnesota (B.S.) and Penn State University (Ph.D.), where he studied chemistry. He completed additional research training at the University of Edinburgh (chemistry), the University of Minnesota (chemistry and electrical engineering), and the Mayo Clinic (neurology). His current research group includes four graduate students and multiple undergraduates who work on a variety of bioanalytical projects. In the new world of “fast track” chemical research, the Wittenberg lab is funded by the NIH for work on high-throughput analysis of microofluidic nanoarrays. Department of Chemistry · Page 3 WITTENBERG LAB USES NIH GRANT TO DEVELOP NEW BIOANALYTICAL TOOLS
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