Integrating a Far-Red Fluorescent Probe with a Microfluidic Platform for Super-Resolution Imaging of Live Erythrocyte Membrane Dynamics

Living erythrocyte (red blood cell, RBC) membranes present rich ultrastructural and dynamic details, which require synchronous super-resolution imaging and single-molecule tracking to be revealed. Yet, it poses a serious challenge to achieve these dual functions in a single probe, due to the rigid and conflicting photophysical demands of the different techniques. Herein, we rationally developed a far-red boron dipyrromethene membrane probe with blinking capability and persistent single-molecule emission, and constructed a microfluidic platform for noninvasive trapping and long-term imaging of RBCs. By combining them, multi-dimensional super-resolution reconstructions and single-molecule tracking were achieved at the molecular scale on living human RBC membranes in a high-throughput manner. Our integrated system defines a quantitative method for analyzing RBC membranes under physiological and pathological conditions, improving precision and revealing new perspectives for future disease diagnostics.

Automated summary

In this study, a new membrane probe and microfluidic platform were developed to enable synchronous use of super-resolution imaging and single-molecule tracking. This method allows high-throughput observation and analysis of the ultrastructural and dynamic details of living red blood cell membranes, providing new perspectives for future disease diagnostics.