Issue 22, 2015

Microfluidic device for DNA amplification of single cancer cells isolated from whole blood by self-seeding microwells

Abstract

Self-seeding microwell chips can sort single cells into 6400 wells based on cell size and their identity verified by immunofluorescence staining. Here, we developed a microfluidic device in which these single cells can be placed, lysed and their DNA amplified for further interrogation. Whole blood spiked with MCF7 tumor cells was passed through the microwell chips after leukocyte depletion and 37% of the MCF7 cells were identified by epithelial cell adhesion molecule (EpCAM) staining in the microwells. Identified single cells were punched into the reaction chamber of the microfluidic device and reagents for cell lysis and DNA amplification introduced sequentially by peristaltic pumping of micro-valves. On-chip lysis and amplification was performed in 8 parallel chambers yielding a 10 000 fold amplification of DNA. Accessibility of the sample through the reaction chamber allowed for easy retrieval and interrogation of target-specific genes to characterize the tumor cells.

Graphical abstract: Microfluidic device for DNA amplification of single cancer cells isolated from whole blood by self-seeding microwells

Supplementary files

Article information

Article type
Paper
Submitted
12 Jul 2015
Accepted
04 Sep 2015
First published
10 Sep 2015

Lab Chip, 2015,15, 4331-4337

Microfluidic device for DNA amplification of single cancer cells isolated from whole blood by self-seeding microwells

Y. Yang, H. S. Rho, M. Stevens, A. G. J. Tibbe, H. Gardeniers and L. W. M. M. Terstappen, Lab Chip, 2015, 15, 4331 DOI: 10.1039/C5LC00816F

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