Full Field Supercritical Angle Fluorescence Microscopy for live cell imaging

TL;DR

This paper presents a novel full field supercritical angle fluorescence microscopy technique that enables real-time, high-resolution imaging of live cell membranes and adhesion phenomena by emission filtering at the sample interface.

Contribution

The authors introduce a new fluorescence imaging method utilizing emission filtering for axial confinement, compatible with standard microscopes, enhancing live cell surface imaging capabilities.

Findings

Achieved ~100 nm axial confinement at the sample interface.

Demonstrated simultaneous imaging of surface and in-depth cell phenomena.

Applicable to live cell membrane and adhesion studies.

Abstract

We introduce a full field fluorescence imaging technique with axial confinement of about 100 nm at the sample/substrate interface. Contrary to standard surface imaging techniques, this confinement is obtained through emission filtering. This technique is based on supercritical emission selectivity. It can be implemented on any epifluorescence microscope with a commercial high numerical aperture objective and offers a real time surface imaging capability. This technique is of particular interest for live cell membrane and adhesion studies. Using HEK cells, we show that one can observe simultaneously the surface and in-depth cell phenomena.