Visible spectrum extended-focus optical coherence microscopy for label-free sub-cellular tomography

Biomed Opt Express. 2017 Jun 20;8(7):3343-3359. doi: 10.1364/BOE.8.003343. eCollection 2017 Jul 1.

Abstract

We present a novel extended-focus optical coherence microscope (OCM) attaining 0.7 μm axial and 0.4 μm lateral resolution maintained over a depth of 40 μm, while preserving the advantages of Fourier domain OCM. Our system uses an ultra-broad spectrum from a supercontinuum laser source. As the spectrum spans from near-infrared to visible wavelengths (240 nm in bandwidth), we call the system visOCM. The combination of such a broad spectrum with a high-NA objective creates an almost isotropic 3D submicron resolution. We analyze the imaging performance of visOCM on microbead samples and demonstrate its image quality on cell cultures and ex-vivo brain tissue of both healthy and alzheimeric mice. In addition to neuronal cell bodies, fibers and plaques, visOCM imaging of brain tissue reveals fine vascular structures and sub-cellular features through its high spatial resolution. Sub-cellular structures were also observed in live cells and were further revealed through a protocol traditionally used for OCT angiography.

Keywords: (000.1430) Biology and medicine; (110.4500) Optical coherence tomography; (170.0180) Microscopy; (170.6900) Three-dimensional microscopy.