Abstract
Three-dimensional high-resolution imaging methods are important for cellular-level research. Optical coherence microscopy (OCM) is a low-coherence-based interferometry technology for cellular imaging with both high axial and lateral resolution. Using a high-numerical-aperture objective, OCM normally has a shallow depth of field and requires scanning the focus through the entire region of interest to perform volumetric imaging. With a higher-numerical-aperture objective, the image quality of OCM is affected by and more sensitive to aberrations. Interferometric synthetic aperture microscopy (ISAM) and computational adaptive optics (CAO) are computed imaging techniques that overcome the depth-of-field limitation and the effect of optical aberrations in optical coherence tomography (OCT), respectively. In this work we combine OCM with ISAM and CAO to achieve high-speed volumetric cellular imaging. Experimental imaging results of ex vivo human breast tissue, ex vivo mouse brain tissue, in vitro fibroblast cells in 3D scaffolds, and in vivo human skin demonstrate the significant potential of this technique for high-speed volumetric cellular imaging.
Highlights
High-speed and high-resolution volumetric imaging is important for many areas of biology and medicine
Interferometric synthetic aperture microscopy (ISAM) tends to smooth the rapid change in the point spread function (PSF) FWHM and provides a much flatter trend around the focus
Dramatic improvement of image quality within the entire imaging volume has been achieved by utilizing the computational optical imaging approaches of ISAM and computational adaptive optics (CAO)
Summary
High-speed and high-resolution volumetric imaging is important for many areas of biology and medicine. High-speed confocal and multiphoton microscopy generate depth-sectioned images by scanning a focus across tissue through a high-numerical-aperture objective lens to achieve superior axial and transverse resolution for cellular imaging. To acquire a three-dimensional (3D) image volume, these techniques require a depth scan after which the different en face images are concatenated to forma so-called z-stack. This method is time consuming, and is susceptible to motion artifacts. In addition to cellular level resolution in both the transverse and axial directions [5,6,7], OCM can achieve better image contrast and greater imaging depth by enhancing the rejection of multiply scattered light over noninterferometric and wide-field methods [8]. There is a well-recognized tradeoff between the lateral resolution and the depth-of-focus
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