To investigate biomolecular dynamics in highly concentrated aqueous environments, we are creating a new platform for near-field, super-resolution optical microscopy by using electron-beam induced cathodoluminescence (CL) of a scintillating thin film as a means to generate a rapidly scannable, nanoscale light source. To non-invasively image molecular complexing, organization, and reorganization at the nanometer scale, our approach is to transfer the robustness and high spatial resolution of scanning electron microscopy (SEM) into a spectrally speci?c and non-invasive optical field in the scintillating film in order to illuminate fluorescently labeled samples in the near-field within a small, adjacent encapsulated volume.To-date, we have generated bright electron beam-induced optical spots that are as small as 18 nm in epitaxially grown thin films of YAlO3:Ce+ and we have demonstrated the ability to image nanoparticles adjacent to the film surface using a variety of forms of contrast with sub-diffraction resolution. We have also demonstrated that these thin films are effective F�rster resonant energy transfer (FRET) donors to dye molecules tethered to their surface. Using low accelerating voltages that limit direct exposure of soft material samples to the electron beam, we expect to resolve the dynamics of membrane protein diffusion and other biomolecular interactions with sub-millisecond pixel dwell times, excellent background rejection, and no moving parts.

Cathodoluminescence-activated non-invasive near-field optical imaging for soft materials at the nanometer scale

248th National Meeting (2014)

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