Surface Plasmon Resonance Imaging of Excitable Cells

Howe, Carmel L.; Webb, Kevin F.; Abayzeed, Sidahmed A.; Anderson, David J.; Denning, Chris; Russell, Noah A.

Abstract

Surface plasmons are highly sensitive to refractive index variations adjacent to the surface. This sensitivity has been exploited successfully for chemical and biological assays. In these systems, a surface plasmon resonance (SPR)-based sensor detects temporal variations in the refractive index at a point. SPR has also been used in imaging systems where the spatial variations of refractive index in the sample provide the contrast mechanism. A high numerical aperture objective lens has been used to design SPR microscopy systems with the ability to image adherent live cells. Addressing research questions in cell physiology and pharmacology often requires the development of a multimodal microscope where complementary information can be obtained.

In this paper, we present the development of a multimodal microscope that combines surface plasmon resonance imaging with a number of additional imaging modalities including bright-field, epi-fluorescence, total internal reflection microscopy (TIRM) and SPR fluorescence microscopy. We used a high numerical aperture objective lens to achieve SPR and TIR microscopy with the ability to image adherent live cells non-invasively. The platform has been used to image live cell cultures demonstrating both fluorescent and label-free techniques. The SPR and TIR imaging systems feature a wide field of view (300 µm) that allows measurements from multiple cells while the resolution is sufficient to image fine cellular processes. The ability of the platform to perform label-free functional imaging of living cell was demonstrated by imaging the spatial variations in contraction of stem cell-derived cardiomyocytes. This technique has a promise for non-invasive imaging of the development of cultured cells over very long periods of time.

• Working Paper Type: Working Paper
• Deposit Date: Apr 20, 2024
• Public URL: https://nottingham-repository.worktribe.com/output/4901458
• Publisher URL: https://www.biorxiv.org/content/10.1101/390948v1