Nanoscopic X-ray tomography for correlative microscopy of a small meiofaunal sea-cucumber.,Scientific Reports

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Nanoscopic X-ray tomography for correlative microscopy of a small meiofaunal sea-cucumber.
Scientific Reports ( IF 3.8 ) Pub Date : 2020-03-03 , DOI: 10.1038/s41598-020-60977-5
Simone Ferstl ₁ , Thomas Schwaha ₂ , Bernhard Ruthensteiner ₃ , Lorenz Hehn ₁ , Sebastian Allner ₁ , Mark Müller ₁ , Martin Dierolf ₁ , Klaus Achterhold ₁ , Franz Pfeiffer _{1,

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Affiliation

In the field of correlative microscopy, light and electron microscopy form a powerful combination for morphological analyses in zoology. Due to sample thickness limitations, these imaging techniques often require sectioning to investigate small animals and thereby suffer from various artefacts. A recently introduced nanoscopic X-ray computed tomography (NanoCT) setup has been used to image several biological objects, none that were, however, embedded into resin, which is prerequisite for a multitude of correlative applications. In this study, we assess the value of this NanoCT for correlative microscopy. For this purpose, we imaged a resin-embedded, meiofaunal sea cucumber with an approximate length of 1 mm, where microCT would yield only little information about the internal anatomy. The resulting NanoCT data exhibits isotropic 3D resolution, offers deeper insights into the 3D microstructure, and thereby allows for a complete morphological characterization. For comparative purposes, the specimen was sectioned subsequently to evaluate the NanoCT data versus serial sectioning light microscopy (ss-LM). To correct for mechanical instabilities and drift artefacts, we applied an alternative alignment procedure for CT reconstruction. We thereby achieve a level of detail on the subcellular scale comparable to ss-LM images in the sectioning plane.

中文翻译：

小型小型动物海参相关显微镜的纳米X射线断层扫描。

在相关显微镜领域，光学显微镜和电子显微镜形成了动物学形态分析的强大组合。由于样本厚度的限制，这些成像技术通常需要切片来研究小动物，从而遭受各种伪影的影响。最近推出的纳米级 X 射线计算机断层扫描 (NanoCT) 装置已用于对多种生物物体进行成像，但这些生物物体均未嵌入树脂中，而树脂是许多相关应用的先决条件。在这项研究中，我们评估了 NanoCT 对于相关显微镜的价值。为此，我们对长度约为 1 毫米的树脂包埋小型动物海参进行了成像，其中 microCT 只能提供很少的内部解剖信息。生成的 NanoCT 数据表现出各向同性 3D 分辨率，提供对 3D 微观结构的更深入了解，从而实现完整的形态表征。出于比较目的，随后对样本进行切片，以评估 NanoCT 数据与连续切片光学显微镜 (ss-LM) 的比较。为了纠正机械不稳定性和漂移伪影，我们应用了 CT 重建的替代对准程序。因此，我们在亚细胞尺度上实现了与切片平面中的 ss-LM 图像相当的细节水平。

更新日期：2020-03-03

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