Soft X-ray tomography to map and quantify organelle interactions at the mesoscale.

Soft X-ray tomography to map and quantify organelle interactions at the mesoscale.
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软X射线层析成像以绘制和量化中尺度上的细胞器相互作用。

DOI:
10.1016/j.str.2022.01.006
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发表时间:
2022-04-07
期刊:
影响因子:
5.7
通讯作者:
Larabell, Carolyn A.
Larabell, Carolyn A.
中科院分区:
生物学2区
文献类型:
--
作者:
Loconte, Valentina;Singla, Jitin;Li, Angdi;Chen, Jian-Hua;Ekman, Axel;McDermott, Gerry;Sali, Andrej;Gros, Mark Le;White, Kate L.;Larabell, Carolyn A.

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细胞器间相互作用是正常细胞功能的重要组成部分;然而,由于细胞生物学中遇到的尺度范围和传统成像方法的通量限制,这些已被证明难以量化。在这里,我们证明了软X射线断层扫描(SXT)可以用来快速映射完整细胞的超微结构重组和细胞器间的相互作用。SXT利用了每个细胞器中富含碳的化合物的天然存在的差异X射线吸收。具体而言,我们使用SXT来绘制胰岛素囊泡的时空演变及其在胰岛素分泌期间和响应于不同刺激时与胰腺β细胞中线粒体的共定位和相互作用。我们量化的形态,生化成分,线粒体和胰岛素囊泡的相对位置的变化。这些发现突出了在中尺度上进行全面和公正的映射以表征细胞重组的重要性,这将难以用其他现有方法检测。软X射线断层扫描结合了快速收集和高分辨率可视化,以了解亚细胞重组。Loconte等人使用该技术来量化和描述胰腺β细胞内细胞器间相互作用的变化,为全面的3D亚细胞定位提供了独特的方法。
Inter-organelle interactions are a vital part of normal cellular function; however, these have proven difficult to quantify due to the range of scales encountered in cell biology and the throughput limitations of traditional imaging approaches. Here, we demonstrate that Soft X-ray Tomography (SXT) can be used to rapidly map ultrastructural reorganization and inter-organelle interactions in intact cells. SXT takes advantage of the naturally occurring, differential X-ray absorption of the carbon-rich compounds in each organelle. Specifically, we use SXT to map the spatio-temporal evolution of insulin vesicles and their co-localization and interaction with mitochondria in pancreatic β-cells during insulin secretion and in response to different stimuli. We quantify changes in the morphology, biochemical composition, and relative position of mitochondria and insulin vesicles. These findings highlight the importance of a comprehensive and unbiased mapping at the mesoscale to characterize cell reorganization that would be difficult to detect with other existing methodologies. Soft X-ray tomography combines rapid collection and high-resolution visualization to understand subcellular reorganization. Loconte et al. use the technique to quantify and describe changes in inter-organelle interactions within pancreatic β-cells, providing a unique approach for comprehensive 3D subcellular mapping.
具有冷冻和X射线的中尺度成像:比分子机大,比单元小。
DOI: 10.1111/boc.201600044
发表时间: 2017-01
影响因子: 2.7
作者:
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发表时间: 2021
影响因子: 5.5
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