High‐throughput microfluidic device for single cell analysis using multiple integrated soft lithographic pumps

Abstract

The ability to accurately control fluid transport in microfluidic devices is key for developing high-throughput methods for single cell analysis. Making small, reproducible changes to flow rates, however, to optimize lysis and injection using pumps external to the microfluidic device are challenging and time-consuming. To improve the throughput and increase the number of cells analyzed, we have integrated previously reported micropumps into a microfluidic device that can increase the cell analysis rate to ∼1000 cells/h and operate for over an hour continuously. In order to increase the flow rates sufficiently to handle cells at a higher throughput, three sets of pumps were multiplexed. These pumps are simple, low-cost, durable, easy to fabricate, and biocompatible. They provide precise control of the flow rate up to 9.2 nL/s. These devices were used to automatically transport, lyse, and electrophoretically separate T-Lymphocyte cells loaded with Oregon green and 6-carboxyfluorescein. Peak overlap statistics predicted the number of fully resolved single-cell electropherograms seen. In addition, there was no change in the average fluorescent dye peak areas indicating that the cells remained intact and the dyes did not leak out of the cells over the 1 h analysis time. The cell lysate peak area distribution followed that expected of an asynchronous steady-state population of immortalized cells.

Monitoring cell secretions on microfluidic chips using solid-phase extraction with mass spectrometry

“…PDMS valves have demonstrated great versatility for on-chip fluid handling, from automating fluid flow over select cell chambers [44] to transporting cells via peristaltic pumping [45]. Pneumatically-actuated valves are integrated on PDMS chips through multilayer assembly [46].…”

Label-free time- and space-resolved exometabolite sampling of growing plant roots through nanoporous interfaces

“…Additional sampling channels and sampling locations can be introduced to enhance the spatiotemporal imaging capability. Moreover, this device can be adapted to simplify fluid pumping and automate process steps by introducing on-chip microfluidic soft lithographic valves 32 . Such an approach could eliminate external tubing and mechanical pumps.…”

“…This PDMS membrane requires a supporting layer, therefore, it was attached to the metabolite sampling-PDMS layer using a multilayer, soft lithographic technique (SI Fig. 1B) 32,35 . The creation of a porous PDMS membrane was based on a previously reported technique 34 .…”

Design and Application of Sensors for Chemical Cytometry

“…Microelectrophoresis chips employing solution or gel-based separations have emerged as a viable alternative to single-cell analysis. ,,,− The microchips offer significant advantages over traditional CE because of their rapid separations (milliseconds to seconds) and are more amenable to parallelization because of their small footprint and very short separation distances . These devices can also be interfaced with detection strategies similar to that used for CE.…”