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Light sheet illumination in single-molecule localization microscopy for imaging of cellular architectures and molecular dynamics

Siyang Cheng, Yuya Nakatani, Gabriella Gagliano, Nahima Saliba, Anna‐Karin Gustavsson

2024npj Imaging11 citationsDOIOpen Access PDF

Abstract

Single-molecule localization microscopy has revealed cellular architectures and molecular dynamics beyond the diffraction limit of light. However, imaging thick samples presents challenges from increased fluorescence background. Light sheet illumination, which utilizes a plane of light for optical sectioning, is effective in reducing fluorescence background, photobleaching, and photodamage. Here, we present the principles of single-molecule localization microscopy and light sheet illumination, followed by an introduction to light sheet microscopy geometries and their imaging applications. Finally, we discuss light sheet illumination approaches for high- and super-resolution imaging of biological structures and dynamics.

Topics & Concepts

Light sheet fluorescence microscopyPhotobleachingMicroscopyPhotoactivated localization microscopyOptical sectioningMaterials scienceFluorescence microscopeSuper-resolution microscopyFluorescenceFluorescence-lifetime imaging microscopyOpticsOptical microscopeNanotechnologyPhysicsScanning electron microscopeAdvanced Fluorescence Microscopy TechniquesAdvanced Electron Microscopy Techniques and ApplicationsAdvanced Biosensing Techniques and Applications
Light sheet illumination in single-molecule localization microscopy for imaging of cellular architectures and molecular dynamics | Litcius