Inertial Multi-Force Deformability Cytometry for High-Throughput, High-Accuracy, and High-Applicability Tumor Cell Mechanotyping.

Previous on-chip technologies for characterizing the cellular mechanical properties often suffer from a low throughput and limited sensitivity. Herein, an inertial multi-force deformability cytometry (IMFDC) is developed for high-throughput, high-accuracy, and high-applicability tumor cell mechanotyping. Three different deformations, including shear deformations and stretch deformations under different forces, are integrated with the IMFDC. The 3D inertial focusing of cells enables the cells to deform by an identical fluid flow, and 10 parameters, such as cell area, perimeter, deformability, roundness, and rectangle deformability, are obtained in three deformations. The IMFDC is able to evaluate the deformability of different cells that are sensitive to different forces on a single chip, demonstrating the high applicability of the IMFDC in analyzing different cell lines. In identifying cell types, the three deformations exhibit different mechanical responses to cells with different sizes and deformability. A discrimination accuracy of ≈93% for both MDA-MB-231 and MCF-10A cells and a throughput of ≈500 cells s-1 can be achieved using the multiple-parameters-based machine learning model. Finally, the mechanical properties of metastatic tumor cells in pleural and peritoneal effusions are characterized, enabling the practical application of the IMFDC in clinical cancer diagnosis.

Cancer cells display increased migration and deformability in pace with metastatic progression.

In this study, we explored the relation between metastatic states vs the capacity of confined migration, amoeboid transition, and cellular stiffness. We compared across an isogenic panel of human breast cancer cells derived from MDA-MB-231 cells. It was observed that cells after lung metastasis have the fastest migration and lowest stiffness, with a significantly higher capacity to transition into an amoeboid mode. Our findings illustrate that metastasis is a selective process favoring motile and softer cells. Moreover, the observation that circulating tumor cells resemble the parental cell line, but not lung-metastatic cells, suggests that cells with higher deformability and motility are likely selected during extravasation and colonization.

High-Throughput Sorting and Single-Cell Mechanotyping by Hydrodynamic Sorting-Mechanotyping Cytometry.

The existence of many background blood cells hinders the accurate identification of circulating tumor cells (CTCs) in the blood of cancer patients. To unlock this limitation, a hydrodynamic sorting-mechanotyping cytometry (HSMC) integrated with a sorting-concentration chip and a detection chip is proposed for simultaneously achieving the high-throughput cell sorting and the multi-parameter mechanotyping of the sorted tumor cells. The HSMC adopts the spiral inertial microfluidics for label-free sorting of cells in a high-throughput manner, allowing the efficient enrichment of tumor cells from the large background blood cells. Then, the sorted cells are concentrated by the concentration unit and finally passed through the detection unit for hydrodynamic deformation. The HSMC has a high throughput for sorting and detection and can successfully reveal the differences in the cellular mechanical properties. After characterizing and optimizing the single chips, the identification of white blood cells (WBCs) and three types of tumor cells (A549, MCF-7, and MDA-MB-231 cells) is successfully achieved. The identification accuracies for WBCs and different tumor cells are all larger than 94%, while the highest identification accuracy is up to 99.2%. This study envisions that the HSMC will offer an avenue for the analysis of single cell intrinsic mechanics in clinical medicine.