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High-contrast multifocus microscopy with a single camera and z-splitter prism.
- Source :
-
Optica [Optica] 2020 Nov 20; Vol. 7 (11), pp. 1477-1486. Date of Electronic Publication: 2020 Oct 22. - Publication Year :
- 2020
-
Abstract
- Optical microscopy has been an indispensable tool for studying complex biological systems, but is often hampered by problems of speed and complexity when performing 3D volumetric imaging. Here, we present a multifocus imaging strategy based on the use of a simple z-splitter prism that can be assembled from off-the-shelf components. Our technique enables a widefield image stack to be distributed onto a single camera and recorded simultaneously. We exploit the volumetric nature of our image acquisition by further introducing a novel extended-volume 3D deconvolution strategy to suppress far-out-of-focus fluorescence background to significantly improve the contrast of our recorded images, conferring to our system a capacity for quasi-optical sectioning. By swapping in different z-splitter configurations, we can prioritize high speed or large 3D field-of-view imaging depending on the application of interest. Moreover, our system can be readily applied to a variety of imaging modalities in addition to fluorescence, such as phase-contrast and darkfield imaging. Because of its simplicity, versatility, and performance, we believe our system will be a useful tool for general biological or biomedical imaging applications.<br />Competing Interests: Disclosures. The authors declare no conflicts of interest.
Details
- Language :
- English
- ISSN :
- 2334-2536
- Volume :
- 7
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- Optica
- Publication Type :
- Academic Journal
- Accession number :
- 34532564
- Full Text :
- https://doi.org/10.1364/optica.404678