Negative depletion mediated brightfield circulating tumour cell identification strategy on microparticle-based microfluidic chip

Abstract

Background: The most convenient circulating tumor cells (CTCs) identification method is direct analysis of cells under bright field microscopy by which CTCs can be comprehensive studied based on morphology, phenotype or even cellular function. However, universal cell markers and a standard tumour cell map do not exist, thus limiting the clinical application of CTCs. Results: This paper focuses on an automatic and convenient negative depletion strategy for circulating tumour cell identification under bright field microscopy. In this strategy, immune microparticles (IMPs) are applied to negatively label white blood cells rather than the tumour cells, such that tumour cells can be directly distinguished under brightfield of the microscopy. In this way, all of the heterogeneous tumour cells and their phenotype properties can be retained for further cancer-related studies. In addition, a wedge-shaped microfluidic chip is constructed for heterogeneous CTC pre-purification and enrichment by size, thus significantly decreasing the interference of haematological cells. Additionally, all cell treatments are processed automatically, and the tumour cells can be rapidly counted and distinguished via customized cell analytical software, showing high detection efficiency and automation. This IMPs based negative cell labelling strategy can also be combined with other classic cell identification methods, thus demonstrating its excellent compatibility. Conclusion: This identification strategy features simple and harmless for tumour cells, as well as excellent accuracy and efficiency. And the low equipment demand and high automation level make it promise for extensive application in basic medical institutions.

Integrated “lab-on-a-chip” microfluidic systems for isolation, enrichment, and analysis of cancer biomarkers

“…Additionally, CTCs isolated through inertial and antibody-capture based methods have been enumerated using optical, electrical and pH-based methods. [88][89][90][91][92][93] One such example is shown in Fig. 2a, where an optofluidic flow cytometer (OFCM) integrating a spiral microfluidic isolation module and fluorescence detection system is used for single cell phenotypic analysis and cell counting.…”

Application of Microfluidics in Detection of Circulating Tumor Cells

“…The desktop on-chip cell sorter is equipped with disposable microfluidic equipment to detect and isolate rare tumor cells for subsequent molecular analysis. Wang et al (2020) used immune microspheres modified with CD45 antibody to label leukocytes. After a wedge chip completes the preliminary sorting of blood cells, CTCs can be directly distinguished under bright-field microscopic imaging, and automatic counting can be realized through image processing software.…”

Combined SERS Microfluidic Chip with Gold Nanocone Array for Effective Early Lung Cancer Prognosis in Mice Model

“…A microfluidic chip is a minor or tens of microns order of magnitude microchannel chip that integrates sample collection, mixing, reaction, separation, and detection operation units, which has become the central implementation platform for immediate detection (POCT). 32 , 33 As a portable instant detection method, the microfluidic chip detection method has the advantages of no pollution, less sample required, and simple operation. It is widely used in disease screening, prevention, postoperative treatment, and other fields.…”