A practical guide to light-sheet microscopy for nanoscale imaging: Looking beyond the cell

被引:2
作者
Kramer, Stephanie N. [1 ]
Antarasen, Jeanpun [1 ]
Reinholt, Cole R. [1 ]
Kisley, Lydia [1 ,2 ]
机构
[1] Case Western Reserve Univ, Dept Phys, Rockefeller Bldg,2076 Adelbert Rd, Cleveland, OH 44106 USA
[2] Case Western Reserve Univ, Dept Chem, Clapp Hall,2080 Adelbert Rd, Cleveland, OH 44106 USA
关键词
PLANE ILLUMINATION MICROSCOPY; FLUORESCENCE MICROSCOPY; CORRELATION SPECTROSCOPY; IN-VIVO; OPTICAL MICROSCOPY; PROTEIN DIFFUSION; RESOLUTION; ENHANCEMENT; DIFFRACTION; EMBRYOS;
D O I
10.1063/5.0218262
中图分类号
O59 [应用物理学];
学科分类号
摘要
We present a comprehensive guide to light-sheet microscopy (LSM) to assist scientists in navigating the practical implementation of this microscopy technique. Emphasizing the applicability of LSM to image both static microscale and nanoscale features, as well as diffusion dynamics, we present the fundamental concepts of microscopy, progressing through beam profile considerations, to image reconstruction. We outline key practical decisions in constructing a home-built system and provide insight into the alignment and calibration processes. We briefly discuss the conditions necessary for constructing a continuous 3D image and introduce our home-built code for data analysis. By providing this guide, we aim to alleviate the challenges associated with designing and constructing LSM systems and offer scientists new to LSM a valuable resource in navigating this complex field.
引用
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页数:24
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