Long-Term Stable and Multifeature Microfluidic Impedance Flow Cytometry Based on a Constricted Channel for Single-Cell Mechanical Phenotyping.

The microfluidic impedance flow cytometer (m-IFC) using constricted microchannels is an appealing choice for the high-throughput measurement of single-cell mechanical properties. However, channels smaller than the cells are susceptible to irreversible blockage, extremely affecting the stability of the system and the throughput. Meanwhile, the common practice of extracting a single quantitative index, i.e., total cell passage time, through the constricted part is inadequate to decipher the complex mechanical properties of individual cells. Herein, this study presents a long-term stable and multifeature m-IFC based on a constricted channel for single-cell mechanical phenotyping. The blockage problem is effectively overcome by adding tiny xanthan gum (XG) polymers. The cells can pass through the constricted channel at a flow rate of 500 μL/h without clogging, exhibiting high throughput (∼240 samples per second) and long-term stability (∼2 h). Moreover, six detection regions were implemented to capture the multiple features related to the whole process of a single cell passing through the long-constricted channel, e.g., creep, friction, and relaxation stages. To verify the performance of the multifeature m-IFC, cells treated with perturbations of microtubules and microfilaments within the cytoskeleton were detected, respectively. It suggests that the extracted features provide more comprehensive clues for single-cell analysis in structural and mechanical transformation. Overall, our proposed multifeature m-IFC exhibits the advantages of nonclogging and high throughput, which can be extended to other cell types for nondestructive and real-time mechanical phenotyping in cost-effective applications.