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[Analytical Chemistry] Towards a droplet radiometric assay for single-cell analysis


Radiotracers are widely used to track molecular processes, both in vitro and in vivo, with high sensitivity and specificity. However, most radionuclide detection methods have spatial resolution inadequate for single-cell analysis. A few existing methods can extract single-cell information from radioactive decays, but the stochastic nature of the process precludes high-throughput measurement (and sorting) of single cells. In this work, we introduce a new concept for translating radioactive decays occurring stochastically within radiolabeled single-cells into an integrated, long-lasting fluorescence signal. Single cells are encapsulated in radiofluorogenic droplets containing molecular probes sensitive to byproducts of ionizing radiation (primarily reactive oxygen species, or ROS). Different probes were examined in bulk solutions, and dihydrorhodamine 123 (DHRh 123) was selected as the lead candidate due to its sensitivity and reproducibility. Fluorescence intensity of DHRh 123 in bulk increased at a rate of 54% per Gy of X-ray radiation and 15% per MBq/ml of 2-deoxy-2-[18F]-fluoro-D-glucose ([18F]FDG). Fluorescence imaging of microfluidic droplets showed the same linear response, but droplets were less sensitive overall than the bulk ROS sensor (detection limit of 3 Gy per droplet). Finally, droplets encapsulating radiolabeled cancer cells allowed, for the first time, the detection of [18F]FDG radiotracer uptake in single cells through fluorescence activation. With further improvements, we expect this technology to enable quantitative measurement and selective sorting of single cells based on the uptake of radiolabeled small molecules.

Maria Elena Gallina, Tae Jin Kim, Mark Shelor, Jaime Vasquez, Amy Mongersun, Minkyu Kim, Sindy K. Y. Tang, Paul Abbyad, and Guillem Pratx Anal. Chem., Just Accepted Manuscript DOI: 10.1021/acs.analchem.7b00414 Publication Date (Web): May 31, 2017 Copyright © 2017 American Chemical Society

Link: http://pubs.acs.org/doi/abs/10.1021/acs.analchem.7b00414

#06032017 #singlecell #dropbasedmicrofluidics

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