Visible light optical coherence microscopy imaging of the mouse cortex with femtoliter volume resolution

Conrad W. Merkle; Shau Poh Chong; Aaron M. Kho; Jun Zhu; Oybek Kholiqov; Alfredo Dubra; Vivek J. Srinivasan

doi:10.1117/12.2292125

3 April 2018 Visible light optical coherence microscopy imaging of the mouse cortex with femtoliter volume resolution

Conrad W. Merkle, Shau Poh Chong, Aaron M. Kho, Jun Zhu, Oybek Kholiqov, Alfredo Dubra, Vivek J. Srinivasan

Proceedings Volume 10483, Optical Coherence Tomography and Coherence Domain Optical Methods in Biomedicine XXII; 104831X (2018) https://doi.org/10.1117/12.2292125
Event: SPIE BiOS, 2018, San Francisco, California, United States

Abstract

Most flying-spot Optical Coherence Tomography (OCT) and Optical Coherence Microscopy (OCM) systems use a symmetric confocal geometry, where the detection path retraces the illumination path starting from and ending with the spatial mode of a single mode optical fiber. Here, we describe a visible light OCM instrument that breaks this symmetry to improve transverse resolution without sacrificing collection efficiency in scattering tissue. This was achieved by overfilling a 0.3 numerical aperture (NA) water immersion objective on the illumination path, while maintaining a conventional Gaussian mode detection path (1/e² intensity diameter ~0.82 Airy disks), enabling ~1.1 μm full-width at half-maximum (FWHM) transverse resolution. At the same time, a ~0.9 μm FWHM axial resolution in tissue, achieved by a broadband visible light source, enabled femtoliter volume resolution. We characterized this instrument according to paraxial coherent microscopy theory, and then used it to image the meningeal layers, intravascular red blood cell-free layer, and myelinated axons in the mouse neocortex in vivo through the thinned skull. Finally, by introducing a 0.8 NA water immersion objective, we improved the lateral resolution to 0.44 μm FWHM, which provided a volumetric resolution of ~0.2 fL, revealing cell bodies in cortical layer I of the mouse brain with OCM for the first time.

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Conrad W. Merkle, Shau Poh Chong, Aaron M. Kho, Jun Zhu, Oybek Kholiqov, Alfredo Dubra, and Vivek J. Srinivasan "Visible light optical coherence microscopy imaging of the mouse cortex with femtoliter volume resolution", Proc. SPIE 10483, Optical Coherence Tomography and Coherence Domain Optical Methods in Biomedicine XXII, 104831X (3 April 2018); https://doi.org/10.1117/12.2292125

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KEYWORDS

Visible radiation

Image resolution

Objectives

Optical coherence tomography

Imaging systems

Confocal microscopy

Tissue optics

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