Complex-field and fluorescence microscopy using aperture scanning technique (CFAST) for studying rhizosphere organisms

Oumeng Zhang; Reinaldo E. Alcalde; Haowen Zhou; Siyuan Yin; Changhuei Yang

doi:10.1117/12.3007534

13 March 2024 Complex-field and fluorescence microscopy using aperture scanning technique (CFAST) for studying rhizosphere organisms

Oumeng Zhang, Reinaldo E. Alcalde, Haowen Zhou, Siyuan Yin, Changhuei Yang

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Proceedings Volume PC12848, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXXI; PC1284802 (2024) https://doi.org/10.1117/12.3007534
Event: SPIE BiOS, 2024, San Francisco, California, United States

Abstract

We design and implement a novel imaging technique that integrates bimodal phase and 3D fluorescence capabilities through aperture segmentation. This approach involves capturing four distinct fluorescence images, mirroring the principles of the Fourier light field microscope and the multi-view reflector microscope, enabling accurate 3D sample reconstruction. Additionally, four brightfield images are acquired for quantitative phase and amplitude reconstruction based on the Kramers-Kronig relations. By combining the strengths of phase imaging, such as digital refocusing, extended depth of field, and non-invasiveness, with the specificity of fluorescence imaging, this method offers a unique imaging solution. Imaging maize roots highlights its exceptional depth of field extension, while imaging a mixture of bacterial cells with and without fluorescent protein tags demonstrates its unique bimodal capabilities.

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Citation Download Citation

Oumeng Zhang, Reinaldo E. Alcalde, Haowen Zhou, Siyuan Yin, and Changhuei Yang "Complex-field and fluorescence microscopy using aperture scanning technique (CFAST) for studying rhizosphere organisms", Proc. SPIE PC12848, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXXI, PC1284802 (13 March 2024); https://doi.org/10.1117/12.3007534

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KEYWORDS

Fluorescence

Fluorescence imaging

Biological imaging

Organisms

3D image processing

Light sources and illumination

Microscopes

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