Special Chemical Engineering Seminar

Monday September 17, 2018 4:00 PM

Ultrafast Single-Cell Level Enzymatic Tumor Profiling

Speaker: Chia-Hung Chen, Department of Biomedical Engineering, National University of Singapore
Location: Spalding Laboratory 106 (Hartley Memorial Seminar Room)

Precision medicine refers to giving the right therapeutics, to the right patient, at the right time. In the context of cancer, successful implementation of precision medicine, requires treatment individualization not only taking into account patient and tumor factors, but also tumor heterogeneity and tumor evolution over time. In this study, a continuous flow microfluidic device was developed as a functional flow cytometer (Drop-Screen) to detect secreted multiplexed protease activities1,2 at single cell resolution (Fig. 1). The individual cells from patient samples are encapsulated within water-in-oil droplets for single cell multiplexed protease assay. We modified FRET (fluorescence resonance energy transfer)-based substrates3 and nano sensors to accommodate different fluorescent pairs with distinct excitation and emission wavelengths to obtain multiple signals from droplets containing single cells. Four substrate-protease reactions in a droplet were simultaneously monitored at three distinct pairs of fluorescent excitation (UV: 400nm, B: 470nm, G: 546nm, R: 635nm) and emission (B: 520nm, G: 580nm, R: 670nm) wavelengths. To infer a quantitative profile of multiple proteolytic activities from single cells, we applied the computational method Proteolytic Activity Matrix Analysis (PrAMA). The capability to determine multiple protease activities at single cell resolution has the potential to characterize tumor progress of individual patients for therapeutics4.

REFERENCES:
[1] E. X. Ng, M. A Miller, T. Jing, D. A Lauffenburger and C. H. Chen, Lab on a Chip, 2015, 15, 1153-1159. 
[2] E. X. Ng, M. A. Miller, T. Jing, and C. H. Chen, Biosensors and Bioelectronics, 2016, 81, 408-414.
[3] M. A. Miller, A. S. Meyer, M. T. Beste, Z. Lasisi, S. Reddy, K. W. Jeng, C. H. Chen, J. Han, K. Isaacson, L. G. Griffith, and D. A. Lauffenburger, PNAS, 2013, E2074–E2083.
[4] H. Tan, S. Guo, D. N. Duy, R. Luo, and C. H. Chen, Nature Communications, 2017, 8, 663.

 

Series Chemical Engineering Seminar

Contact: Sohee Lee at 626-395-4193 sohee@cheme.caltech.edu