Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection
Scanning light-sheet microscopy in the whole mouse brain with HiLo background rejection
复制标题
DOI：
10.1117/1.3324890
复制
发表时间：
2010-01-01
影响因子：
3.5
通讯作者：
Kim, Jinhyun
中科院分区：
文献类型：
--
作者：
Mertz, Jerome;Kim, Jinhyun
众所周知，光片照明可以实现宏观样品的光学切片宽视场成像。然而，光片宏观的光学切片能力受到样品引起的散射或像差的破坏，这些散射或像差会扩大片照度的厚度。提出了一种利用失焦背景抑制增强扫描光片显微镜光学切片能力的方法。这项技术被称为HiLo显微镜，它利用均匀和有结构的薄片照明连续获得的两幅图像。然后通过融合来自两个图像的高、低空间频率信息合成光学切片图像。使用绿色荧光蛋白(GFP)-荧光和暗场散射光对比，在光学清除的全小鼠脑样品中证明了结合光片宏观观察和HiLo背景抑制的好处。(C) 2010光学仪器工程师学会。(DOI: 10.1117/1.3324890)
It is well known that light-sheet illumination can enable optically sectioned wide-field imaging of macroscopic samples. However, the optical sectioning capacity of a light-sheet macroscope is undermined by sample-induced scattering or aberrations that broaden the thickness of the sheet illumination. We present a technique to enhance the optical sectioning capacity of a scanning light-sheet microscope by out-of-focus background rejection. The technique, called HiLo microscopy, makes use of two images sequentially acquired with uniform and structured sheet illumination. An optically sectioned image is then synthesized by fusing high and low spatial frequency information from both images. The benefits of combining light-sheet macroscopy and HiLo background rejection are demonstrated in optically cleared whole mouse brain samples, using both green fluorescent protein (GFP)-fluorescence and dark-field scattered light contrast. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3324890]