A generalizable and targeted molecular biopsy approach for in situ cryogenic electron tomography of vitreous brain tissue.

IF 4.5 Q1 BIOCHEMICAL RESEARCH METHODS

Cell Reports Methods Pub Date : 2025-07-21 Epub Date: 2025-06-16 DOI:10.1016/j.crmeth.2025.101080

Calina Glynn, Jake L R Smith, Matthew Case, Rebecca Csöndör, Ana Katsini, Maria E Sanita, Thomas S Glen, Avery Pennington, Michael Grange

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Abstract

Cellular cryogenic electron tomography (cryo-ET) enables the capture of detailed structural information within a biologically relevant environment. However, information in more complex samples, such as multicellular specimens and tissues, is lacking. Importantly, these observations need to be set in the context of populations. Currently, imaging on the molecular scale is limited to a few observations in situ that struggle to be generalized. This is due to limitations in throughput and versatility employed by current instrumentation. Here, we utilize plasma focused ion beam milling to examine the molecular landscape of mouse hippocampus by cryo-ET. We reveal the complex organization of macromolecules in targeted regions across CA1 stratum pyramidale (sp) to radiatum (sr), representing a molecular atlas of hippocampal architecture in adult mice. The combination of instrumentation and application of technical advancements provides a framework to explore specific structural questions within other tissues in a targeted manner.

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玻璃体脑组织原位低温电子断层扫描的一种可推广的靶向分子活检方法。

细胞低温电子断层扫描（cryo-ET）能够在生物相关环境中捕获详细的结构信息。然而，在更复杂的样本，如多细胞标本和组织的信息，是缺乏的。重要的是，这些观察需要在人口的背景下进行。目前，分子尺度上的成像仅限于一些原位观察，难以推广。这是由于当前仪器在吞吐量和通用性方面的限制。在这里，我们利用等离子体聚焦离子束铣削，通过冷冻- et来研究小鼠海马的分子景观。我们揭示了大分子在CA1金字塔层（sp）到辐射层（sr）的目标区域的复杂组织，代表了成年小鼠海马结构的分子图谱。仪器和技术进步的应用相结合提供了一个框架，以有针对性的方式探索其他组织中的特定结构问题。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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