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Speaker: John P. Nolan Ph. D., La Jolla Bioengineering Institute.

Single cell analysis is recognized as a key tool in understanding how the complex cell systems that form tissues, organs, and tumors operate. Improving this understanding is critically dependent on new methods to analyze single cells with high molecular specificity, multiplexibility sensitivity, and speed. Our lab is developing instrumentation, reagents, and assays to better study and exploit nanoscale phenomenon related to cell function. We use optical methods extensively, especially flow cytometry, and recently developed the first high resolution Spectral Flow Cytometers. These instruments use high efficiency gratings and high speed detectors to measure the complete emission spectra of individual cells at rates of hundreds of cells per second. Spectral flow cytometry produce data comparable or superior to conventional flow cytometers for many common applications, but also opens a door to new applications. As an example, we have developed a set of nanoparticle surface enhanced Raman scattering (SERS) tags that can be used as labels for antibodies or other targeting molecules. As many as 20 distinct SERS tags can be discriminated based on spectral features contained within ~100 nm of the spectrum. Combined with fluorescence probes, the use of SERS tags can significantly increase the level of multiplexing possible compared with conventional instruments. We are also interested in the quantitative analysis of the extracellular vesicles (exosomes, ectosomes, and other microvesicles) released by cells, and have developed a high sensitivity Nanoparticle Flow Cytometer and associated methods that allow us to size and count individual membrane vesicles as small as 50 nm in diameter. These new methods are being applied to the development of biomarkers for cardiovascular toxicity.

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