Microfluidic active loading of single cells enables analysis of complex clinical specimens.
Nicholas L CalistriRobert J KimmerlingSeth W MalinowskiMehdi TouatMark M StevensSelim OlcumKeith L LigonScott R ManalisPublished in: Nature communications (2018)
A fundamental trade-off between flow rate and measurement precision limits performance of many single-cell detection strategies, especially for applications that require biophysical measurements from living cells within complex and low-input samples. To address this, we introduce 'active loading', an automated, optically-triggered fluidic system that improves measurement throughput and robustness by controlling entry of individual cells into a measurement channel. We apply active loading to samples over a range of concentrations (1-1000 particles μL-1), demonstrate that measurement time can be decreased by up to 20-fold, and show theoretically that performance of some types of existing single-cell microfluidic devices can be improved by implementing active loading. Finally, we demonstrate how active loading improves clinical feasibility for acute, single-cell drug sensitivity measurements by deploying it to a preclinical setting where we assess patient samples from normal brain, primary and metastatic brain cancers containing a complex, difficult-to-measure mixture of confounding biological debris.
Keyphrases
- single cell
- rna seq
- high throughput
- induced apoptosis
- living cells
- squamous cell carcinoma
- small cell lung cancer
- fluorescent probe
- resting state
- emergency department
- stem cells
- endoplasmic reticulum stress
- label free
- single molecule
- functional connectivity
- circulating tumor cells
- intensive care unit
- signaling pathway
- subarachnoid hemorrhage
- brain injury
- loop mediated isothermal amplification