Single-cell proteolytic activity measurement using microfluidics for rare cell populations

Yu Chih Chen, Euisik Yoon

Research output: Chapter in Book/Report/Conference proceedingChapter


Proteolytic degradation of the extracellular matrix represents a key step in cancer dissemination and metastasis. To probe cellular proteolytic activity, fluorescent sensing substrate was developed, yet prior studies focused on average activity of thousands of cells. Considerable evidence suggests a specialized subset of cancer cells are driving metastasis, highlighting the value of single-cell approach to reveal cancer cellular heterogeneity. In addition, when only a small number of cells are available, single-cell analysis is required to draw a statistical conclusion. Here, we present a microfluidic platform that provides high-efficiency cell loading and simple valveless isolation, so the proteolytic activity of a small number (10–100) of cells can be individually characterized. Furthermore, the platform allows monitoring single cells at multiple time points for the investigation of dynamics in proteolytic activity. The presented platform represents a simple and reliable tool for single-cell proteolytic analysis, illuminating the heterogeneous and dynamic nature of cancer cells.

Original languageEnglish
Title of host publicationMethods in Enzymology
EditorsNancy L. Allbritton, Michelle L. Kovarik
PublisherAcademic Press Inc.
Number of pages15
ISBN (Print)9780128170908
Publication statusPublished - 2019

Publication series

NameMethods in Enzymology
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Bibliographical note

Funding Information:
This work was supported by the grants R01 CA 203810 and R21 CA 195016 from NIH to E.Y. The support of Y.-C.C. from Forbes Institute for Cancer Discovery is also acknowledged. We thank the Lurie Nanofabrication Facility of the University of Michigan (Ann Arbor, MI) for device fabrication.

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

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