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ultraLM和miniLM：用于在相关光电子显微镜中智能追踪荧光细胞的定位工具。

ultraLM and miniLM: Locator tools for smart tracking of fluorescent cells in correlative light and electron microscopy.

作者信息

Brama Elisabeth, Peddie Christopher J, Wilkes Gary, Gu Yan, Collinson Lucy M, Jones Martin L

机构信息

Electron Microscopy Science Technology Platform, The Francis Crick Institute, London, UK.

出版信息

Wellcome Open Res. 2016 Dec 13;1:26. doi: 10.12688/wellcomeopenres.10299.1.

DOI:10.12688/wellcomeopenres.10299.1
PMID:28090593
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5234702/
Abstract

In-resin fluorescence (IRF) protocols preserve fluorescent proteins in resin-embedded cells and tissues for correlative light and electron microscopy, aiding interpretation of macromolecular function within the complex cellular landscape. Dual-contrast IRF samples can be imaged in separate fluorescence and electron microscopes, or in dual-modality integrated microscopes for high resolution correlation of fluorophore to organelle. IRF samples also offer a unique opportunity to automate correlative imaging workflows. Here we present two new locator tools for finding and following fluorescent cells in IRF blocks, enabling future automation of correlative imaging. The ultraLM is a fluorescence microscope that integrates with an ultramicrotome, which enables 'smart collection' of ultrathin sections containing fluorescent cells or tissues for subsequent transmission electron microscopy or array tomography. The miniLM is a fluorescence microscope that integrates with serial block face scanning electron microscopes, which enables 'smart tracking' of fluorescent structures during automated serial electron image acquisition from large cell and tissue volumes.

摘要

树脂内荧光（IRF）方案可在树脂包埋的细胞和组织中保存荧光蛋白，用于相关光镜和电镜检查，有助于在复杂的细胞环境中解释大分子功能。双对比IRF样本可在单独的荧光显微镜和电子显微镜中成像，也可在双模态集成显微镜中成像，以实现荧光团与细胞器的高分辨率关联。IRF样本还为自动化相关成像工作流程提供了独特的机会。在这里，我们展示了两种新的定位工具，用于在IRF块中查找和跟踪荧光细胞，从而实现未来相关成像的自动化。ultraLM是一种与超薄切片机集成的荧光显微镜，可对含有荧光细胞或组织的超薄切片进行“智能采集”，以便后续进行透射电子显微镜检查或阵列断层扫描。miniLM是一种与连续块面扫描电子显微镜集成的荧光显微镜，可在从大细胞和组织体积自动采集连续电子图像期间对荧光结构进行“智能跟踪”。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/6ca30ca7e901/wellcomeopenres-1-11092-g0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/17b7b60e9255/wellcomeopenres-1-11092-g0000.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/f554afcf912a/wellcomeopenres-1-11092-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/1cc4c92f9d53/wellcomeopenres-1-11092-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/706b89a31e21/wellcomeopenres-1-11092-g0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/db3d00e415be/wellcomeopenres-1-11092-g0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/4e3918fb1834/wellcomeopenres-1-11092-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/84b29af3cfb8/wellcomeopenres-1-11092-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/6ca30ca7e901/wellcomeopenres-1-11092-g0007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/17b7b60e9255/wellcomeopenres-1-11092-g0000.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/f554afcf912a/wellcomeopenres-1-11092-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/1cc4c92f9d53/wellcomeopenres-1-11092-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/706b89a31e21/wellcomeopenres-1-11092-g0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/db3d00e415be/wellcomeopenres-1-11092-g0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/4e3918fb1834/wellcomeopenres-1-11092-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/84b29af3cfb8/wellcomeopenres-1-11092-g0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cddd/5446011/6ca30ca7e901/wellcomeopenres-1-11092-g0007.jpg

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