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Optical microscopy has vastly expanded the frontiers of structural and functional biology, due to the non-invasive probing of dynamic volumes in vivo. However, traditional widefield microscopy illuminating the entire field of view (FOV) is adversely affected by out-of-focus light scatter. Consequently, standard upright or inverted microscopes are inept in sampling diffraction-limited volumes smaller than the optical system’s point spread function (PSF). Over the last few decades, several planar and structured (sinusoidal) illumination modalities have offered unprecedented access to sub-cellular organelles and 4D (3D + time) image acquisition. Furthermore, these optical sectioning systems remain unaffected by the size of biological samples, providing high signal-to-noise (SNR) ratios for objective lenses (OLs) with long working distances (WDs). This review aims to guide biologists regarding planar illumination strategies, capable of harnessing sub-micron spatial resolution with a millimeter depth of penetration.
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Novel Image Processing to Restore Scattered Light-sheet Microscopic Imaging Technique and its Application for Quantifying Biomechanics
Research of cellular and molecular processes by way of histological methods allows for some insight but comes with a fundamental set of constraints that are challenging to overcome. Traditional histological methods are laborious, as well as severely limiting for in-depth study of developmental processes or disease processes in vivo. In traditional histology, fixing and sectioning tissue necessarily eliminates its dynamic function, while tissue section thickness limits the scope of investigation with conventional imaging tools. Noninvasive in vivo study of tissues and biomarkers is therefore paramount in gaining a fuller understanding of the pathophysiology surrounding conditions like congenital heart disorders. Light-sheet fluorescence microscopy (LSFM) is a powerful and noninvasive optical microscopy tool that can image in vivo tissue function in 4D (3D + time). LSFM boasts benefits such as short pixel dwell time (and therefore minimal photobleaching) while maintaining the ability to image a high dynamic range, as well as deep-tissue optical sectioning. Researchers have been seeking to overcome this problem by developing tissue-clearing techniques to attempt to homogenize the refractive index across the tissue via the removal of light-scattering pigments and lipids.
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- Award ID(s):
- 1936519
- PAR ID:
- 10498262
- Publisher / Repository:
- River Publishers
- Date Published:
- ISBN:
- 9788770223799
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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