Scattered‐light‐sheet microscopy with sub‐cellular resolving power

Scattered‐light‐sheet microscopy with sub‐cellular resolving power
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具有亚细胞分辨率的散射光片显微镜

DOI:
10.1002/jbio.202300068
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发表时间:
2023
影响因子:
2.8
通讯作者:
Vasdekis, Andreas E.
Vasdekis, Andreas E.
中科院分区:
物理与天体物理2区
文献类型:
--
作者:
Subedi, Nava R.;Stolyar, Sergey;Tuson, Sabrina J.;Marx, Christopher J.;Vasdekis, Andreas E.

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自100多年前首次演示以来,基于散射的光片显微镜最近重新成为无标签组织成像和细胞形态测量的关键方式;然而,基于散射的亚细胞分辨率光片成像仍然是一个未实现的目标。这是因为相关方法不可避免地将斑点或颗粒强度调制叠加到原生亚细胞特征上。在这里,我们通过部署时间平均的伪热化光片照明来解决这一挑战。虽然这种方法增加了照明片的横向尺寸,但我们在图像反卷积后实现了亚细胞分辨率。我们通过对酵母和细菌的细胞质碳库进行成像验证了这种方法,该方法具有更高的特异性,无染色和超低辐照水平。总的来说,我们期望这种基于散射的光片显微镜方法能够通过低辐照度和无标签操作来消除光毒性,从而推进单个活细胞成像。
Since its first demonstration over 100 years ago, scattering‐based light‐sheet microscopy has recently re‐emerged as a key modality in label‐free tissue imaging and cellular morphometry; however, scattering‐based light‐sheet imaging with subcellular resolution remains an unmet target. This is because related approaches inevitably superimpose speckle or granular intensity modulation on to the native subcellular features. Here, we addressed this challenge by deploying a time‐averaged pseudo‐thermalized light‐sheet illumination. While this approach increased the lateral dimensions of the illumination sheet, we achieved subcellular resolving power after image deconvolution. We validated this approach by imaging cytosolic carbon depots in yeast and bacteria with increased specificity, no staining, and ultralow irradiance levels. Overall, we expect this scattering‐based light‐sheet microscopy approach will advance single, live cell imaging by conferring low‐irradiance and label‐free operation towards eradicating phototoxicity.
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