Abstract
Rapid increase in interest and applications of through-focus (TF) or volumetric type of optical imaging in biology and other areas has resulted in the development of several TF image collection methods. Achieving quantitative results from images requires standardization and optimization of image acquisition protocols. Several standardization protocols are available for conventional optical microscopy where a best-focus image is used, but to date, rigorous testing protocols do not exist for TF optical imaging. In this paper, we present a method to determine the fidelity of the TF optical data using the TF scanning optical microscopy images.
Highlights
Through-focus (TF) optical imaging is steadily gaining momentum in several areas such as biological imaging, optical metrology, microscopy, adaptive optics, material processing, optical data storage, and optical inspection [1,2,3,4,5,6,7,8,9,10,11,12,13,14]
In a first attempt to standardize TF image collection protocols, here we presented a method to determine the fidelity of the TF optical images using TF scanning optical microscopy (TSOM)
We studied the effect of random lateral and axial vibrations on the resulting noise as evaluated by differential TSOM (D-TSOM) images
Summary
Through-focus (TF) optical imaging is steadily gaining momentum in several areas such as biological imaging, optical metrology, microscopy, adaptive optics, material processing, optical data storage, and optical inspection [1,2,3,4,5,6,7,8,9,10,11,12,13,14]. TF imaging includes extended-depth-of-field, blurred, defocused, three-dimensional (3D), extended-focused, outof-focus, axial scanning, and volumetric imaging. TF imaging is used to acquire images of brain tissue and bone calcium in three dimensions [4, 7, 8]. High-speed TF imaging has been reported to track single-molecules in three dimensions [28], to image the entire embryos [29], and to track the 3D dynamics in live cells [5, 6, 30, 31]. Cellular network dynamics was demonstrated using TF imaging in three dimensions [7]. Cellular network dynamics was demonstrated using TF imaging in three dimensions [7]. 3D automated nanoparticle tracking was demonstrated using TF images [32]
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