1-D High-Throughput Screening Assays for Primary Human T Cells

Recorded On: 02/05/2018

The parallel microfluidic cytometer (PMC) is an imaging flow cytometer that operates on statistical analysis of low-pixel-count, one-dimensional (1-D) line scans. It is efficient in data collection and operates on suspension cells. Our 1-D instrument leverages both the high throughput aspects of traditional flow cytometry and the high spatial content of 2-D imaging cytometers. In this talk, we present a supervised automated pipeline for the PMC that minimizes operator intervention by incorporating automated multivariate logistic regression for data scoring. The approach quantifies biomarker localization of activated T cells into a single descriptive ‘activity score’ readout. Reducing complex phenotypes into a simple readout has many advantages for drug screening and characterization. We test the self-tuning statistical algorithms in human primary T cells in flow with various drug response assays. We readily achieve an average Z’ of 0.55 and SSMD of 13. The PMC is volume efficient, needing only 4 µL of sample volume per well. Anywhere from 3000 to 9000 independent sample tests can be processed from a single 15 mL blood donation. The parallel nature of our laser scanning system enables high well throughput and is extremely scalable. We conclude that the new technology will support primary cell protein localization assays and “on-the-fly” data scoring at a sample throughput of more than 100,000 wells per day. This is, in principle, consistent with large-scale primary pharmaceutical screens. We demonstrate that 1-D imaging provides many advantages for rapid development of primary T cell assays in flow.

Steve Wang

Boston University

Previous research assistant in Daniel Ehrlich's group at Boston University. Developed 1-D cytometry methods and high throughput automated statistical approaches to screening in primary T cells.

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