Issue 2, 2016

Microfluidic compartments with sensing microbeads for dynamic monitoring of cytokine and exosome release from single cells

Abstract

Monitoring activity of single cells has high significance for basic science and diagnostic applications. Here we describe a reconfigurable microfluidic device for confining single cells along with antibody-modified sensing beads inside 20 picoliter (pL) microcompartments for monitoring cellular secretory activity. An array of ∼7000 microchambers fabricated in the roof of the reconfigurable microfluidic device could be raised or lowered by applying negative pressure. The floor of the device was micropatterned to contain cell attachment sites in registration with the microcompartments. Using this set-up, we demonstrated the detection of inflammatory cytokine IFN-γ and exosomes from single immune cells and cancer cells respectively. The detection scheme was similar in both cases: cells were first captured on the surface inside the microfluidic device, then sensing microbeads were introduced into the device so that, once the microcompartments were lowered, single cells and microbeads became confined together. The liquid bathing the beads and the cells inside the compartments also contained fluorescently-labeled secondary antibodies (Abs). The capture of cell-secreted molecules onto microbeads was followed by binding of secondary antibodies – this caused microbeads to become fluorescent. The fluorescence intensity of the microbeads changed over time, providing dynamics of single cell secretory activity. The microdevice described here may be particularly useful in the cases where panning upstream of sensing is required or to analyze secretory activity of anchorage-dependent cells.

Graphical abstract: Microfluidic compartments with sensing microbeads for dynamic monitoring of cytokine and exosome release from single cells

Supplementary files

Article information

Article type
Paper
Submitted
13 ኦገስ 2015
Accepted
04 ኦክቶ 2015
First published
05 ኦክቶ 2015

Analyst, 2016,141, 679-688

Microfluidic compartments with sensing microbeads for dynamic monitoring of cytokine and exosome release from single cells

K. J. Son, A. Rahimian, D. Shin, C. Siltanen, T. Patel and A. Revzin, Analyst, 2016, 141, 679 DOI: 10.1039/C5AN01648G

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements