Clinical Validation of an Ultra High-Throughput Spiral Microfluidics for the Detection and Enrichment of Viable Circulating Tumor Cells

Background: Circulating tumor cells (CTCs) are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation. Methodology/Principal Findings: Here, we enriched and characterized putative CTCs from blood samples of patients with both advanced stage metastatic breast and lung cancers using a novel multiplexed spiral microfluidic chip. This system detected putative CTCs under high sensitivity (100%, n = 56) (Breast cancer samples: 12–1275 CTCs/ml; Lung cancer samples: 10–1535 CTCs/ml) rapidly from clinically relevant blood volumes (7.5 ml under 5 min). Blood samples were completely separated into plasma, CTCs and PBMCs components and each fraction were characterized with immunophenotyping (Pan-cytokeratin/CD45, CD44/CD24, EpCAM), fluorescence in-situ hybridization (FISH) (EML4-ALK) or targeted somatic mutation analysis. We used an ultra-sensitive mass spectrometry based system to highlight the presence of an EGFR-activating mutation in both isolated CTCs and plasma cell-free DNA (cf-DNA), and demonstrate concordance with the original tumor-biopsy samples. Conclusions/Significance: We have clinically validated our multiplexed microfluidic chip for the ultra high-throughput, low-cost and label-free enrichment of CTCs. Retrieved cells were unlabeled and viable, enabling potential propagation and real-time downstream analysis using next generation sequencing (NGS) or proteomic analysis.Singapore-MIT Alliance for Research and Technolog

Continuous particle separation in spiral microchannels using dean flows and differential migration

Microparticle separation and concentration based on size has become indispensable in many biomedical and environmental applications. In this paper we describe a passive microfluidic device with spiral microchannel geometry for complete separation of particles. The design takes advantage of the inertial lift and viscous drag forces acting on particles of various sizes to achieve differential migration, and hence separation, of microparticles. The dominant inertial forces and the Dean rotation force due to the spiral microchannel geometry cause the larger particles to occupy a single equilibrium position near the inner microchannel wall. The smaller particles migrate to the outer half of the channel under the influence of Dean forces resulting in the formation of two distinct particle streams which are collected in two separate outputs. This is the first demonstration that takes advantage of the dual role of Dean forces for focusing larger particles in a single equilibrium position and transposing the smaller particles from the inner half to the outer half of the microchannel cross-section. The 5-loop spiral microchannel 100 mm wide and 50 mm high was used to successfully demonstrate a complete separation of 7.32 mm and 1.9 mm particles at Dean number De = 0.47. Analytical analysis supporting the experiments and models is also presented. The simple planar structure of the separator offers simple fabrication and makes it ideal for integration with on-chip microfluidic systems, such as micro total analysis systems (mTAS) or lab-on-a-chip (LOC) for continuous filtration and separation applications

Single cell kinase signaling assay using pinched flow coupled droplet microfluidics

10.1063/1.4878635Biomicrofluidics833410

Addressing cellular heterogeneity in tumor and circulation for refined prognostication

10.1073/pnas.1907904116PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA1163617957-1796

Wearable Soft Microtube Sensors for Quantitative Home-Based Erectile Dysfunction Monitoring

Quantifiable erectile dysfunction (ED) diagnosis involves the monitoring of rigidity and tumescence of the penile shaft during nocturnal penile tumescence (NPT). In this work, we introduce Erectile Dysfunction SENsor (EDSEN), a home-based wearable device for quantitative penile health monitoring based on stretchable microtubular sensing technology. Two types of sensors, the T- and R-sensors, are developed to effectively measure penile tumescence and rigidity, respectively. Conical models mimicking penile shaft were fabricated with polydimethylsiloxane (PDMS) material, using different base to curing agent ratios to replicate the different hardness properties of a penile shaft. A theoretical buckling force chart for the different penile models is generated to determine sufficiency criteria for sexual intercourse. An average erect penile length and circumference requires at least a Young’s modulus of 179 kPa for optimal buckling force required for satisfactory sexual intercourse. The conical penile models were evaluated using EDSEN. Our results verified that the circumference of a penile shaft can be accurately measured by T-sensor and rigidity using the R-sensor. EDSEN provides a private and quantitative method to detect ED within the comfortable confines of the user’s home

A microfluidics approach towards high-throughput pathogen removal from blood using margination

Sepsis is an adverse systemic inflammatory response caused by microbial infection in blood. This paper reports a simple microfluidic approach for intrinsic, non-specific removal of both microbes and inflammatory cellular components (platelets and leukocytes) from whole blood, inspired by the invivo phenomenon of leukocyte margination. As blood flows through a narrow microchannel (20 × 20 µm), deformable red blood cells (RBCs) migrate axially to the channel centre, resulting in margination of other cell types (bacteria, platelets, and leukocytes) towards the channel sides. By using a simple cascaded channel design, the blood samples undergo a 2-stage bacteria removal in a single pass through the device, thereby allowing higher bacterial removal efficiency. As an application for sepsis treatment, we demonstrated separation of Escherichia coli and Saccharomyces cerevisiae spiked into whole blood, achieving high removal efficiencies of ∼80% and ∼90%, respectively. Inflammatory cellular components were also depleted by >80% in the filtered blood samples which could help to modulate the host inflammatory response and potentially serve as a blood cleansing method for sepsis treatment. The developed technique offers significant advantages including high throughput (∼1 ml/h per channel) and label-free separation which allows non-specific removal of any blood-borne pathogens (bacteria and fungi). The continuous processing and collection mode could potentially enable the return of filtered blood back to the patient directly, similar to a simple and complete dialysis circuit setup. Lastly, we designed and tested a larger filtration device consisting of 6 channels in parallel (∼6 ml/h) and obtained similar filtration performances. Further multiplexing is possible by increasing channel parallelization or device stacking to achieve higher throughput comparable to convectional blood dialysis systems used in clinical settings

Erratum: Integrative analysis and machine learning based characterization of single circulating tumor cells (J. Clin. Med., (2020) 9, 1206, 10.3390/jcm9041206)

10.3390/jcm10020370Journal of Clinical Medicine1021-Fe

Erratum: Iyer, A., et al. Integrative Analysis and Machine Learning Based Characterization of Single Circulating Tumor Cells. <em>J. Clin. Med.</em> 2020, <em>9</em>, 1206

In the published manuscript [...