Visual Quantitative Detection of Circulating Tumor Cells with Single-Cell Sensitivity Using a Portable Microfluidic Device.

Immunocytological technologies, molecular technologies, and functional assays are widely used for detecting circulating tumor cells (CTCs) after enrichment from patients' blood sample. Unfortunately, accessibility to these technologies is limited due to the need for sophisticated instrumentation and skilled operators. Portable microfluidic devices have become attractive tools for expanding the access and efficiency of detection beyond hospitals to sites near the patient. Herein, a volumetric bar chart chip (V-Chip) is developed as a portable platform for CTC detection. The target CTCs are labeled with aptamer-conjugated nanoparticles (ACNPs) and analyzed by V-Chip through quantifying the byproduct (oxygen) of the catalytic reaction between ACNPs and hydrogen peroxide, which results in the movement of an ink bar to a concentration-dependent distance for visual quantitative readout. Thus, the CTC number is decoded into visually quantifiable information and a linear correlation can be found between the distance moved by the ink and number of cells in the sample. This method is sensitive enough that a single cell can be detected. Furthermore, the clinical capabilities of this system are demonstrated for quantitative CTC detection in the presence of a high leukocyte background. This portable detection method shows great potential for quantification of rare cells with single-cell sensitivity for various applications.

Isolation, Detection, and Antigen-Based Profiling of Circulating Tumor Cells Using a Size-Dictated Immunocapture Chip.

Even though the diagnostic and prognostic value of circulating tumor cells (CTCs) has been demonstrated, their clinical utility and widespread adoption have been limited. Herein, we describe a new device, size-dictated immunocapture chip (SDI-Chip), for efficient, sensitive, and spatially resolved capture and detection of CTCs. SDI-Chip enables selective, frequent, and extended interaction of CTCs with hydrodynamically optimized immunocoated micropillar surfaces. CTCs with different antigen expression levels can be efficiently captured and spatially resolved around the micropillars. Capture efficiency greater than 92 % with a purity of 82 % was achieved with blood samples. CTCs were detected in non-metastasis colorectal (CRC) patients, while none was detected from healthy volunteers. We believe that SDI-Chip will facilitate the transition of tumor diagnosis from anatomical pathology to molecular pathology in localized CRC patients.

Distance-based paper/PMMA integrated ELISA-chip for quantitative detection of immunoglobulin G.

The enzyme-linked immunosorbent assay (ELISA) is one of the most commonly implemented clinical diagnostic tools for the detection and quantification of protein biomarkers. However, conventional ELISA tests require sophisticated infrastructure, expensive reagents, long assay time, and expertise for operation, which are not often easily accessible in resource-limited settings. Microfluidic ELISA-chip based point-of-care (POC) testing allows miniaturization and integration of complex functions that facilitate their usage in limited-resource settings. The current work demonstrates a simple, portable, low cost and equipment-free paper/poly(methyl methacrylate) (PMMA) integrated microfluidic ELISA-chip as a POC device with a visual distance-based readout for quantitative detection of clinical biomarkers. The integrated paper/PMMA ELISA-chip utilizes the movement of immunoassay complexes with magnetic beads by a permanent magnet in a PMMA part of the compartment. The target concentration is translated into a visual distance signal readout for quantitative detection of biomarkers in a µPAD. Because it does not require sophisticated instruments and has the added advantages of low cost, easy operation, and disposability with quantitative visual readout, the paper/PMMA ELISA-chip holds great promise for portable detection of target bioanalytes as a POC diagnostic tool in resource-limited setups.