New Generation of Ensemble-Decision Aliquot Ranking Based on Simplified Microfluidic Components for Large-Capacity Trapping of Circulating Tumor Cells

Abstract

Ensemble-decision aliquot ranking (eDAR) is a sensitive and high-throughput method to analyze circulating tumor cells (CTCs) from peripheral blood. Here, we report the next generation of eDAR, where we designed and optimized a new hydrodynamic switching scheme for the active sorting step in eDAR, which provided fast cell sorting with an improved reproducibility and stability. The microfluidic chip was also simplified by incorporating a functional area for subsequent purification using microslits fabricated by standard lithography method. Using the reported second generation of eDAR, we were able to analyze 1-mL of whole-blood samples in 12.5 min, with a 95% recovery and a zero false positive rate (n=15).

Optical painting and fluorescence activated sorting of single adherent cells labelled with photoswitchable Pdots

“…eDAR was designed and demonstrated to have high recovery efficiency (95%) for the isolation of rare cells from whole blood. It offers high sensitivity and high throughput: the method can process 1 ml of whole blood containing ∼5 billion red and white blood cells within 20 min (refs 39 , 40 ). Compared with a commercial FACS instrument, which often requires large amounts of sample cells entering it (10 6 –10 7 ) (ref.…”

Microfluidic applications on circulating tumor cell isolation and biomimicking of cancer metastasis

“…A number of microfluidics‐based technologies have been developed for CTC isolation and biomimicking of tumor microenvironment to evaluate cancer metastasis. Although the capture efficiency can reach 98% with purity of 89% [87] and throughput can reach 3 mL/min [75] for CTC isolation using microfluidic technologies, the performance of these methods is not good enough and still needs further improvement. To better meet the clinical requirements of CTC detection, the technology should tend to the following demands: (a) higher isolation efficiency which may need new technologies or combination of several technologies; (b) specificity verification strategies to prove that the isolated targeted cells are real CTCs which have clinical significance; (c) accuracy and repeatability that apply automated apparatus as much as possible without much intervention from human operators; (d) operability and generalizability; (e) fully heterogeneity consideration to distinguish different CTC subpopulations; and (f) clinical demonstration to prove this technology is useful in clinical trials.…”

Circulating tumor microemboli: Progress in molecular understanding and enrichment technologies

“…CTM have also been detected using another automated high throughput enrichment free CTC identification/enumeration system, namely eDAR (ensemble-decision aliquot ranking) (Schiro et al, 2012;Zhao et al, 2013). Unprocessed blood samples are labeled with fluorescent antibodies against CTC specific cell surface markers (e.g.…”

“…Fluorescent anti-EpCAM (and/or-HER2) labeling, laser illumination and APD detection Isolation of CTCs/CTM for further analysis through (i) track etched polycarbonate membrane filtration, or (ii) Microfabricated slits Many CTM with low EpCAM expression Throughput: 1 ml blood/12.5 min (Schiro et al, 2012;Zhao et al, 2013) Abbreviations: PCA: Prostate Cancer (Ozkumur et al, 2013) CTM differentially express a subset of genes compared to CTCs Plakoglobin upregulated in CTM and its knock-out disintegrates CTM Abundance of CTM denote adverse outcomes in breast cancer pateints (Aceto et al, 2014)…”