Dual-Wavelength Surface Plasmon Resonance Microscopy Combined with Laser-Induced Bubble-Cell Perforation: A Novel Single-Cell Manipulation and Real-Time Monitoring Platform

Abstract

Precise single-cell membrane perforation holds great promise in membrane biology, yet reported approaches face two major challenges: (1) a lack of both spatiotemporal precision and efficiency at a single-cell resolution; (2) current methods cannot noninvasively monitor the real-time perforation dynamics. Herein, we propose a novel methodology for controlling cellular membrane perforation and real-time monitoring of mechanobiological imaging with single-cell resolution. The proposed methodology was developed by an advanced dual-wavelength surface plasmon resonance microscopy platform combined with femtosecond laser-induced microbubble generation, enabling us to achieve a precise spatial profile in membrane perturbation through laser focal positioning. Moreover, a typical dual-wavelength fitting algorithm was employed to determine the resonance wavelength (RW), providing a subsecond temporal resolution of 0.1 s/frame. More importantly, the label-free imaging method can provide three key advantages: (1) Noninvasive monitoring of membrane dynamics via an adhesion-dependent RW mapping; (2) accurate and controllable perforation at a single-cell level; and (3) low-cost configuration. The integrated platform can establish an important framework for noninvasive investigation of the dynamic process of the cell membrane under controllable external stimulation in real-time. It can be expected that this advancement in the live-cell imaging field can offer a versatile analytical platform for performing fundamental membrane biophysics studies.