Real-time detection of lipid bilayer assembly and detergent-initiated solubilization using optical cavities.

Abstract

The cellular membrane governs numerous fundamental biological processes. Therefore, developing a comprehensive understanding of its structure and function is critical. However, its inherent biological complexity gives rise to numerous inter-dependent physical phenomena. In an attempt to develop a model, two different experimental approaches are being pursued in parallel: performing single cell experiments (top down) and using biomimetic structures (bottom up), such as lipid bilayers. One challenge in many of these experiments is the reliance on fluorescent probes for detection which can create confounds in this already complex system. In the present work, a label-free detection method based on an optical resonant cavity is used to detect one of the fundamental physical phenomena in the system: assembly and solubilization of the lipid bilayer. The evanescent field of the cavity strongly interacts with the lipid bilayer, enabling the detection of the bilayer behavior in real-time. Two independent detection mechanisms confirm the formation and detergent-assisted solubilization of the lipid bilayers: (1) a refractive index change and (2) a material loss change. Both mechanisms can be monitored in parallel, on the same device, thus allowing for cross-confirmation of the results. To verify the proposed method, we have detected the formation of self-assembled phosphatidylcholine lipid bilayers from small unilamellar vesicles on the device surface in real-time. Subsequently, we exposed the bilayers to two different detergents (non-ionic Triton X-100 and anionic sodium dodecyl sulfate) to initiate solubilization, and this process was also detected in real-time. After the bilayer solubilization, the device returned to its initial state, exhibiting minimal hysteresis. The experimental wash-off was also collected and analyzed using dynamic light scattering.