An Enzyme‐Loaded Metal–Organic Framework‐Assisted Microfluidic Platform Enables Single‐Cell Metabolite Analysis

Abstract

Colonization of cancer cells at secondary sites, a decisive step in tumor metastasis, is strongly dependent on the formation of metastatic microenvironments regulated by intrinsic single‐cell metabolism traits. Herein, we report a single‐cell microfluidic platform for high‐throughput dynamic monitoring of tumor cell metabolites to evaluate tumor malignancy. This microfluidic device empowers efficient isolation of single cells (>99 %) in a squashed state similar to tumor extravasation, and employs enzyme‐packaged metal–organic frameworks to catalyze tumor cell metabolites for visualization. The microfluidic evaluation was confirmed by in vivo assays, suggesting that the platform allowed predicting the tumorigenicity of captured tumor cells and screening metabolic inhibitors as anti‐metastatic drugs. Furthermore, the platform efficiently detected various aggressive cancer cells in unprocessed whole blood samples with high sensitivity, showing potential for clinical application.

Electrochemiluminescence Analysis of Multiple Glycans on Single Living Cell with a Closed Bipolar Electrode Array Chip

“…Microfluidic chips allow for accurate manipulation of single cells, such as trapping, releasing, and moving of cells by electrical, mechanical, or optical means . In recent years, there have been significant advances in innovative strategies for both cell isolation and related biomarker analysis at the single-cell level. − For example, Li et al introduced a revolutionary label-free optofluidic nanoplasmonic biosensor, capable of real-time single-cell analysis .…”

“…In addition, the heterogeneity of the single cells before and after drug treatment were also investigated. Gao et al developed a single-cell microfluidic platform that efficiently isolated individual tumor cells (>99% isolation rate) by replicating the squashed state during tumor extravasation . The platform employed enzyme-packaged metal–organic frameworks to catalyze tumor cell metabolites, enabling their visualization.…”

“…3 The combination of effective single-cell manipulation and in situ analysis of biomarkers offers a powerful tool for advancing our understanding of biomarker-related biological processes and translating this knowledge into clinical applications for the diagnosis and therapy of tumor-related diseases. 4,5 Microfluidic chips allow for accurate manipulation of single cells, such as trapping, 6 releasing, 7 and moving of cells by electrical, 8 mechanical, 9 or optical means. 10 In recent years, there have been significant advances in innovative strategies for both cell isolation and related biomarker analysis at the singlecell level.…”

“…Gao isolation rate) by replicating the squashed state during tumor extravasation. 6 The platform employed enzyme-packaged metal−organic frameworks to catalyze tumor cell metabolites, enabling their visualization. Although these strategies have successfully accomplished single-cell isolation and biomarker detection, tumor identification was not accurate enough solely relying on a single biomarker due to the insufficient information and high false positive signal.…”

Centrifugo-Pneumatic Reciprocating Flowing Coupled with a Spatial Confinement Strategy for an Ultrafast Multiplexed Immunoassay

“…In the past few decades, numerous efforts have been made to tackle the aforementioned challenges by harnessing emerging microfluidic technology. In contrast to conventional manual operation and static incubation reactions, the microfluidic approach involves integrating traditional immune-analysis procedures onto a microchip and simplifying the process through continuous flow-based reactions. ,− Notably, the ultralow dimensions of the microfluidic platform contribute to a high surface-to-volume ratio, thereby reducing diffusion distance. − However, it is important to consider that the single-pass flow-through scheme employed in most microfluidics can lead to the waste of unbound antibodies. Additionally, the antibodies present in the reaction solution usually have only one chance to interact with the antigens coated on the substrate. , While this approach reduces time consumption, it may potentially compromise the efficiency of antigen–antibody reactions.…”