机械拉伸细胞的高压冷冻。

IF 1.5 4区 工程技术 Q3 MICROSCOPY
Edward Felder, Jan L Rüth, Bassam Abu-Omar, Martin Wohlwend, Paul Walther, Clarissa Read
{"title":"机械拉伸细胞的高压冷冻。","authors":"Edward Felder, Jan L Rüth, Bassam Abu-Omar, Martin Wohlwend, Paul Walther, Clarissa Read","doi":"10.1111/jmi.13411","DOIUrl":null,"url":null,"abstract":"<p><p>High-pressure freezing (HPF) is an electron microscopy (EM) preparation technique with superb ultrastructural preservation. Combined with EM tomography it provides virtual EM serial sections with extraordinary spatial resolution. For HPF, cells are usually cultured on a rigid sapphire disc that provides a tight fit in the holding bracket of the HPF apparatus. Since we are using extensible elastic silicone membranes as a growth support to perform cell stretch experiments, we developed a method to clamp the stretched silicone membrane and place it instead of the sapphire disc into the HPF holding bracket. Compared to chemical fixation the HPF immobilised cells showed improved structural preservation, partly even on a molecular level. However, the outstanding quality of HPF immobilised cells on sapphire discs was not achieved. Moreover, regions with obvious freezing artefacts seemed to be more abundant in the HPF silicone membranes, probably caused by lower heat transfer rates of the silicone membrane during the HPF process. Taken together, we have shown that HPF immobilisation can be performed on growth supports different than sapphire discs. Since even stretched membranes can be used with the new method, also other unconventional growth supports should not pose a problem.</p>","PeriodicalId":16484,"journal":{"name":"Journal of microscopy","volume":" ","pages":""},"PeriodicalIF":1.5000,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"High-pressure freezing of mechanically stretched cells.\",\"authors\":\"Edward Felder, Jan L Rüth, Bassam Abu-Omar, Martin Wohlwend, Paul Walther, Clarissa Read\",\"doi\":\"10.1111/jmi.13411\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>High-pressure freezing (HPF) is an electron microscopy (EM) preparation technique with superb ultrastructural preservation. Combined with EM tomography it provides virtual EM serial sections with extraordinary spatial resolution. For HPF, cells are usually cultured on a rigid sapphire disc that provides a tight fit in the holding bracket of the HPF apparatus. Since we are using extensible elastic silicone membranes as a growth support to perform cell stretch experiments, we developed a method to clamp the stretched silicone membrane and place it instead of the sapphire disc into the HPF holding bracket. Compared to chemical fixation the HPF immobilised cells showed improved structural preservation, partly even on a molecular level. However, the outstanding quality of HPF immobilised cells on sapphire discs was not achieved. Moreover, regions with obvious freezing artefacts seemed to be more abundant in the HPF silicone membranes, probably caused by lower heat transfer rates of the silicone membrane during the HPF process. Taken together, we have shown that HPF immobilisation can be performed on growth supports different than sapphire discs. Since even stretched membranes can be used with the new method, also other unconventional growth supports should not pose a problem.</p>\",\"PeriodicalId\":16484,\"journal\":{\"name\":\"Journal of microscopy\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":1.5000,\"publicationDate\":\"2025-04-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of microscopy\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1111/jmi.13411\",\"RegionNum\":4,\"RegionCategory\":\"工程技术\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"MICROSCOPY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of microscopy","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1111/jmi.13411","RegionNum":4,"RegionCategory":"工程技术","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"MICROSCOPY","Score":null,"Total":0}
引用次数: 0

摘要

高压冷冻(HPF)是一种具有优良超微结构保存性能的电子显微镜制备技术。结合电磁断层成像,它提供了具有非凡空间分辨率的虚拟电磁序列切片。对于HPF,细胞通常在坚硬的蓝宝石圆盘上培养,该圆盘与HPF设备的支架紧密配合。由于我们使用可扩展的弹性硅膜作为生长支撑来进行细胞拉伸实验,我们开发了一种方法来夹紧拉伸的硅膜,并将其代替蓝宝石盘放入HPF支架中。与化学固定相比,HPF固定的细胞显示出更好的结构保存,部分甚至在分子水平上。然而,在蓝宝石圆盘上的HPF固定细胞的卓越质量并没有实现。此外,具有明显冷冻伪影的区域似乎在HPF硅膜中更为丰富,这可能是由于在HPF过程中硅膜的传热速率较低所致。综上所述,我们已经证明HPF固定可以在不同于蓝宝石盘的生长支架上进行。因为即使拉伸膜也可以与新方法一起使用,其他非常规的生长支撑也不会造成问题。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
High-pressure freezing of mechanically stretched cells.

High-pressure freezing (HPF) is an electron microscopy (EM) preparation technique with superb ultrastructural preservation. Combined with EM tomography it provides virtual EM serial sections with extraordinary spatial resolution. For HPF, cells are usually cultured on a rigid sapphire disc that provides a tight fit in the holding bracket of the HPF apparatus. Since we are using extensible elastic silicone membranes as a growth support to perform cell stretch experiments, we developed a method to clamp the stretched silicone membrane and place it instead of the sapphire disc into the HPF holding bracket. Compared to chemical fixation the HPF immobilised cells showed improved structural preservation, partly even on a molecular level. However, the outstanding quality of HPF immobilised cells on sapphire discs was not achieved. Moreover, regions with obvious freezing artefacts seemed to be more abundant in the HPF silicone membranes, probably caused by lower heat transfer rates of the silicone membrane during the HPF process. Taken together, we have shown that HPF immobilisation can be performed on growth supports different than sapphire discs. Since even stretched membranes can be used with the new method, also other unconventional growth supports should not pose a problem.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Journal of microscopy
Journal of microscopy 工程技术-显微镜技术
CiteScore
4.30
自引率
5.00%
发文量
83
审稿时长
1 months
期刊介绍: The Journal of Microscopy is the oldest journal dedicated to the science of microscopy and the only peer-reviewed publication of the Royal Microscopical Society. It publishes papers that report on the very latest developments in microscopy such as advances in microscopy techniques or novel areas of application. The Journal does not seek to publish routine applications of microscopy or specimen preparation even though the submission may otherwise have a high scientific merit. The scope covers research in the physical and biological sciences and covers imaging methods using light, electrons, X-rays and other radiations as well as atomic force and near field techniques. Interdisciplinary research is welcome. Papers pertaining to microscopy are also welcomed on optical theory, spectroscopy, novel specimen preparation and manipulation methods and image recording, processing and analysis including dynamic analysis of living specimens. Publication types include full papers, hot topic fast tracked communications and review articles. Authors considering submitting a review article should contact the editorial office first.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信