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
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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.

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单分子定位显微镜中的光片照明,用于细胞结构和分子动力学成像
单分子定位显微镜揭示了超越光衍射极限的细胞结构和分子动力学。然而,厚样品成像会因荧光背景增加而面临挑战。光片照明利用光平面进行光学切片,可有效减少荧光背景、光漂白和光损伤。在此,我们将介绍单分子定位显微镜和光片照明的原理,然后介绍光片显微镜的几何结构及其成像应用。最后,我们将讨论用于生物结构和动力学高分辨率和超分辨率成像的光片照明方法。
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