Subcellular three-dimensional imaging deep through multicellular thick samples by structured illumination microscopy and adaptive optics.

Subcellular three-dimensional imaging deep through multicellular thick samples by structured illumination microscopy and adaptive optics.
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通过结构化照明显微镜和自适应光学器件,通过多细胞厚样品深入细胞的三维成像。

DOI:
10.1038/s41467-021-23449-6
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发表时间:
2021-05-25
影响因子:
16.6
通讯作者:
Kner P
Kner P
中科院分区:
综合性期刊1区
文献类型:
--
作者:
Lin R;Kipreos ET;Zhu J;Khang CH;Kner P

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结构照明显微镜使实时成像与亚衍射分辨率。不幸的是,光学像差会导致结构照明显微镜分辨率和伪影的损失,使得该技术在比单个细胞厚的样品中无法使用。在这里,我们报道了自适应光学和结构照明显微镜的结合,在80µm深度下,通过秀丽隐杆线虫以150 nm的横向分辨率和570 nm的轴向分辨率成像。我们证明了自适应光学提高了三维分辨率,特别是沿轴向,并减少了伪影,成功地在各种生物样品中实现了三维结构照明显微镜。结构照明显微镜(SIM)中的光学像差会导致分辨率损失和伪影,使其不适合厚样品。本文将自适应光学与SIM (AO-3DSIM)相结合,提高了三维分辨率,减少了伪影,对线虫进行了3D-SIM。
Structured Illumination Microscopy enables live imaging with sub-diffraction resolution. Unfortunately, optical aberrations can lead to loss of resolution and artifacts in Structured Illumination Microscopy rendering the technique unusable in samples thicker than a single cell. Here we report on the combination of Adaptive Optics and Structured Illumination Microscopy enabling imaging with 150 nm lateral and 570 nm axial resolution at a depth of 80 µm through Caenorhabditis elegans. We demonstrate that Adaptive Optics improves the three-dimensional resolution, especially along the axial direction, and reduces artifacts, successfully realizing 3D-Structured Illumination Microscopy in a variety of biological samples. Optical aberrations in Structured Illumination Microscopy (SIM) can lead to loss of resolution and artifacts making it unsuitable for thick samples. Here the authors combine Adaptive Optics and SIM (AO-3DSIM) to improve the 3D resolution and reduce artifacts, performing 3D-SIM in C.elegans.
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