• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

用于哺乳动物细胞冷冻深剖样品的冷冻-FM 和冷冻-FIB-SEM 相关的样品制备和图像配准。

Sample preparation and image registration for correlative cryo-FM and cryo-FIB-SEM of plunge-frozen mammalian cells.

机构信息

Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.

Department of Chemical Research Support, Weizmann Institute of Science, Rehovot, Israel.

出版信息

STAR Protoc. 2022 Feb 8;3(1):101142. doi: 10.1016/j.xpro.2022.101142. eCollection 2022 Mar 18.

DOI:10.1016/j.xpro.2022.101142
PMID:35199027
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8844706/
Abstract

We recently demonstrated how lipid droplets can serve as fiducials for correlating cryo-fluorescence microscopy (cryo-FM) and cryo-focused ion beam scanning electron microscopy (cryo-FIB-SEM) datasets of mammalian cells grown on grids. Here we describe a step-by-step protocol for correlative cryo-FM and cryo-FIB-SEM, starting from sample preparation of C2C12 cell line, followed by imaging with cryo-FM and cryo-FIB-SEM. Finally, we detail how to perform the 3D-correlation with sub-micron accuracy. For complete details on the use and execution of this profile, please refer to Scher et al. (2021).

摘要

我们最近展示了如何将脂滴用作在网格上生长的哺乳动物细胞的冷冻荧光显微镜 (cryo-FM) 和冷冻聚焦离子束扫描电子显微镜 (cryo-FIB-SEM) 数据集相关联的基准。在这里，我们描述了一个从 C2C12 细胞系的样品制备开始，然后进行 cryo-FM 和 cryo-FIB-SEM 成像的分步协议。最后，我们详细介绍了如何以亚微米精度执行 3D 相关。有关此配置文件的使用和执行的完整详细信息，请参阅 Scher 等人。（2021 年）。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/41aee14ca31f/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/e7feff47271a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/8eec6a1e2cd5/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/66b00b40808d/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/c81384fa8bee/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/31f26e98e2e9/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/3c4206f89795/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/07e3726ad1a3/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/41aee14ca31f/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/e7feff47271a/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/8eec6a1e2cd5/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/66b00b40808d/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/c81384fa8bee/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/31f26e98e2e9/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/3c4206f89795/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/07e3726ad1a3/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e49c/8844706/41aee14ca31f/gr7.jpg

相似文献

1
Sample preparation and image registration for correlative cryo-FM and cryo-FIB-SEM of plunge-frozen mammalian cells.用于哺乳动物细胞冷冻深剖样品的冷冻-FM 和冷冻-FIB-SEM 相关的样品制备和图像配准。
STAR Protoc. 2022 Feb 8;3(1):101142. doi: 10.1016/j.xpro.2022.101142. eCollection 2022 Mar 18.
2
Advanced cryo-tomography workflow developments - correlative microscopy, milling automation and cryo-lift-out.先进的冷冻电子断层扫描工作流程发展——相关显微镜技术、铣削自动化和冷冻提取技术。
J Microsc. 2021 Feb;281(2):112-124. doi: 10.1111/jmi.12939. Epub 2020 Jul 2.
3
Sample Preparation by 3D-Correlative Focused Ion Beam Milling for High-Resolution Cryo-Electron Tomography.通过 3D 相关聚焦离子束铣削进行高分辨率冷冻电子断层成像的样品制备。
J Vis Exp. 2021 Oct 25(176). doi: 10.3791/62886.
4
A cryogenic, coincident fluorescence, electron, and ion beam microscope.低温共焦荧光、电子和离子束显微镜。
Elife. 2022 Oct 28;11:e82891. doi: 10.7554/eLife.82891.
5
A focused ion beam milling and lift-out approach for site-specific preparation of frozen-hydrated lamellas from multicellular organisms.一种用于从多细胞生物体中进行特定部位冷冻水合薄片制备的聚焦离子束铣削和剥离方法。
J Struct Biol. 2015 Nov;192(2):262-9. doi: 10.1016/j.jsb.2015.07.012. Epub 2015 Jul 26.
6
fiducial markers for 3D correlative cryo-fluorescence and FIB-SEM imaging.用于三维相关冷冻荧光和聚焦离子束扫描电子显微镜成像的基准标记
iScience. 2021 Jun 10;24(7):102714. doi: 10.1016/j.isci.2021.102714. eCollection 2021 Jul 23.
7
A modular platform for automated cryo-FIB workflows.一个用于自动化 cryo-FIB 工作流程的模块化平台。
Elife. 2021 Dec 24;10:e70506. doi: 10.7554/eLife.70506.
8
An introduction to cryo-FIB-SEM cross-sectioning of frozen, hydrated Life Science samples.介绍用于冷冻、水合生命科学样品的冷冻-FIB-SEM 切片技术。
J Microsc. 2021 Feb;281(2):138-156. doi: 10.1111/jmi.12951. Epub 2020 Aug 24.
9
PIE-scope, integrated cryo-correlative light and FIB/SEM microscopy.PIE-scope，集成冷冻相关的光和 FIB/SEM 显微镜。
Elife. 2019 Jul 1;8:e45919. doi: 10.7554/eLife.45919.
10
Site-Specific Cryo-focused Ion Beam Sample Preparation Guided by 3D Correlative Microscopy.由三维相关显微镜引导的位点特异性冷冻聚焦离子束样品制备
Biophys J. 2016 Feb 23;110(4):860-9. doi: 10.1016/j.bpj.2015.10.053. Epub 2016 Jan 5.

本文引用的文献

1
fiducial markers for 3D correlative cryo-fluorescence and FIB-SEM imaging.用于三维相关冷冻荧光和聚焦离子束扫描电子显微镜成像的基准标记
iScience. 2021 Jun 10;24(7):102714. doi: 10.1016/j.isci.2021.102714. eCollection 2021 Jul 23.
2
Protocol for image registration of correlative soft X-ray tomography and super-resolution structured illumination microscopy images.相关软 X 射线断层扫描和超分辨率结构光照明显微镜图像的配准协议。
STAR Protoc. 2021 May 6;2(2):100529. doi: 10.1016/j.xpro.2021.100529. eCollection 2021 Jun 18.
3
Cryo-FIB-SEM as a promising tool for localizing proteins in 3D.
Cryo-FIB-SEM 作为一种在 3D 中定位蛋白质的有前途的工具。
J Struct Biol. 2020 Jul 1;211(1):107528. doi: 10.1016/j.jsb.2020.107528. Epub 2020 May 6.
4
eC-CLEM: flexible multidimensional registration software for correlative microscopies.eC-CLEM：用于相关显微镜技术的灵活多维配准软件。
Nat Methods. 2017 Jan 31;14(2):102-103. doi: 10.1038/nmeth.4170.
5
Cryo-FIB-SEM serial milling and block face imaging: Large volume structural analysis of biological tissues preserved close to their native state.冷冻聚焦离子束扫描电子显微镜连续铣削和块面成像：对接近自然状态保存的生物组织进行大体积结构分析。
J Struct Biol. 2016 Dec;196(3):487-495. doi: 10.1016/j.jsb.2016.09.016. Epub 2016 Sep 28.
6
New hardware and workflows for semi-automated correlative cryo-fluorescence and cryo-electron microscopy/tomography.用于半自动相关低温荧光和低温电子显微镜/断层扫描的新硬件和工作流程。
J Struct Biol. 2017 Feb;197(2):83-93. doi: 10.1016/j.jsb.2016.06.020. Epub 2016 Jun 28.
7
Cryo FIB-SEM: volume imaging of cellular ultrastructure in native frozen specimens.冷冻 FIB-SEM：对天然冷冻标本中细胞超微结构的体积成像。
J Struct Biol. 2013 Nov;184(2):355-60. doi: 10.1016/j.jsb.2013.09.024. Epub 2013 Oct 9.
8
Icy: an open bioimage informatics platform for extended reproducible research.Icy：一个开放的生物影像信息学平台，用于扩展可重复研究。
Nat Methods. 2012 Jun 28;9(7):690-6. doi: 10.1038/nmeth.2075.
9
Fiji: an open-source platform for biological-image analysis.斐济：一个用于生物影像分析的开源平台。
Nat Methods. 2012 Jun 28;9(7):676-82. doi: 10.1038/nmeth.2019.
10
Nile red: a selective fluorescent stain for intracellular lipid droplets.尼罗红：一种用于细胞内脂滴的选择性荧光染料。
J Cell Biol. 1985 Mar;100(3):965-73. doi: 10.1083/jcb.100.3.965.