Journal
LAB ON A CHIP
Volume 15, Issue 7, Pages 1656-1666Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c5lc00036j
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Funding
- Harper Cancer Research Institute
- Walther Cancer Foundation
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There has been increasing evidence that micro and messenger RNA derived from exosomes play important roles in pancreatic and other cancers. In this work, a microfluidics-based approach to the analysis of exosomal RNA is presented based on surface acoustic wave (SAW) exosome lysis and ion-exchange nanomembrane RNA sensing performed in conjunction on two separate chips. Using microRNA hsa-miR-550 as a model target and raw cell media from pancreatic cancer cell lines as a biological sample, SAW-based exosome lysis is shown to have a lysis rate of 38%, and an ion-exchange nanomembrane sensor is shown to have a limit of detection of 2 pM, with two decades of linear dynamic range. A universal calibration curve was derived for the membrane sensor and used to detect the target at a concentration of 13 pM in a SAWlysed sample, which translates to 14 target miRNA per exosome from the raw cell media. At a total analysis time of similar to 1.5 h, this approach is a significant improvement over existing methods that require two overnight steps and 13 h of processing time. The platform also requires much smaller sample volumes than existing technology (similar to 100 mu L as opposed to similar to mL) and operates with minimal sample loss, a distinct advantage for studies involving mouse models or other situations where the working fluid is scarce.
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