The Department of Chemistry and Physics continues its seminar series. Details for this week’s talk are given below.
Friday, 11/18/2022
3-4pm Eastern
Kristopher Amrhein
UTC Biochem ‘17
PhD Candidate
Department of Chemistry, University of Memphis
Grote Hall 411
Dual Imaging Single Vesicle Technology for Exosome Characterization and Early Cancer Detection
Abstract
Single exosome molecular profiling has the potential to transform clinical cancer detection and monitoring by precisely probing cancer-associated exosomes in the presence of normal exosomes at the single vesicle level, but it is a grand challenge because of the small exosome size, low abundance of antigens on individual exosomes, and complex biological matrix. Here, we report a clinically practical dual imaging single vesicle technology (DISVT) for surface protein profiling of individual exosomes in biofluids without pre-purification. The DISVT is based on a facile dual imaging platform, which molecularly captures exosomes directly from diluted biofluids, localizes individual exosomes with membrane dye and fluorescence imaging with angled laser excitation, and detect targeted surface protein markers on the individual exosomes with surface plasmon light scattering gold nanoparticles and dark field imaging. A fast semi-automatic method was developed for dual image analysis using Bash scripts, Python, and ImageJ. Using DISVT, we profiled exosomes derived from different breast cancer cell lines and plasma exosomes from healthy donors and HER2-positive breast cancer patients at early and locally advantaged stages. The results show that the DISVT, but not the traditional enzyme-linked immunosorbent assay (ELISA), detected breast cancer at early-stage, raising exciting possibilities of single exosome protein profiling as a new tool for early cancer detection.
Keenan E. Dungey, PhD
Department Head, Chemistry & Physics