Abstract
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.
Highlights
IntroductionPlanar Illumination Strategies for Optical tomography has contributed tremendously to facilitating the non-invasive, quantitative modeling of tissue homeostasis and biochemical/mechanical mechanisms [1,2,3,4,5,6,7]
This review aims to cover recently developed planar illumination strategies that have enabled the rapid acquisition of subcellular volumes, hidden behind the optical barrier
Combined with tissue-clearing or expansion methods, Light sheet microscopy (LSM) allows for the opportunity to investigate molecular structure and function across different scales that were previously unable to be reconstructed without aberrations induced by tissue
Summary
Planar Illumination Strategies for Optical tomography has contributed tremendously to facilitating the non-invasive, quantitative modeling of tissue homeostasis and biochemical/mechanical mechanisms [1,2,3,4,5,6,7]. A tradeoff is observed between spatio-temporal resolution, fluorescence labeling density and laser power required to avoid photo bleaching for a millimeter depth of penetration [56,59,73,76,77,78,79,80] In this regard, planar illumination has successfully filled the niche for dynamic, subcellular fluorescence tomography. LSM has been used to visualize complex neuronal processes or dendritic spine pathophysiology in situ Such subcellular processes are routinely subject to noise and/or poor framerates for confocal laser scanning and widefield microscopy [11,12,14]. This review aims to cover recently developed planar illumination strategies that have enabled the rapid acquisition of subcellular volumes, hidden behind the optical barrier
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
