Challenges and limitations of molecular resolution fluorescence imaging.

IF 2.4 3区 化学 Q3 CHEMISTRY, ANALYTICAL
Dominic A Helmerich, Markus Sauer
{"title":"Challenges and limitations of molecular resolution fluorescence imaging.","authors":"Dominic A Helmerich, Markus Sauer","doi":"10.1088/2050-6120/ae042b","DOIUrl":null,"url":null,"abstract":"<p><p>Super-resolution microscopy (SRM) has revolutionized fluorescence imaging enabling insights into the molecular organization of cells that were previously unconceivable. Latest developments now allow the visualization of individual molecules with nanometer precision and imaging with molecular resolution. However, translating these achievements to imaging under physiological conditions in cells remains challenging. The higher the spatial resolution is pushed by the development of improved SRM methods the more challenging the problems we are confronted when aiming to use them for sub-10 nm fluorescence imaging in cells. It turns out that most developed SRM methods that demonstrate nanometer resolution cannot be directly implemented for molecular resolution imaging in cells. Particularly, fluorescence labeling, i.e. high-density covalent labeling of the molecules of interest with fluorophores with minimal linkage error represents currently a nearly insurmountable obstacle. In addition, even if high labeling densities can be realized it has to be considered that fluorophores can interact via different energy pathways and thus impede super-resolution imaging in the sub-10 nm range. Here, we describe the boundaries, discuss the challenges we must accept and show strategies to circumvent them and achieve true molecular resolution fluorescence imaging under physiological conditions in cells.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":2.4000,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods and Applications in Fluorescence","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1088/2050-6120/ae042b","RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0

Abstract

Super-resolution microscopy (SRM) has revolutionized fluorescence imaging enabling insights into the molecular organization of cells that were previously unconceivable. Latest developments now allow the visualization of individual molecules with nanometer precision and imaging with molecular resolution. However, translating these achievements to imaging under physiological conditions in cells remains challenging. The higher the spatial resolution is pushed by the development of improved SRM methods the more challenging the problems we are confronted when aiming to use them for sub-10 nm fluorescence imaging in cells. It turns out that most developed SRM methods that demonstrate nanometer resolution cannot be directly implemented for molecular resolution imaging in cells. Particularly, fluorescence labeling, i.e. high-density covalent labeling of the molecules of interest with fluorophores with minimal linkage error represents currently a nearly insurmountable obstacle. In addition, even if high labeling densities can be realized it has to be considered that fluorophores can interact via different energy pathways and thus impede super-resolution imaging in the sub-10 nm range. Here, we describe the boundaries, discuss the challenges we must accept and show strategies to circumvent them and achieve true molecular resolution fluorescence imaging under physiological conditions in cells.

分子分辨率荧光成像的挑战和局限性。
超分辨率显微镜(SRM)彻底改变了荧光成像,使人们能够深入了解以前无法想象的细胞分子组织。最新的发展现在允许单个分子的可视化与纳米精度和成像与分子分辨率。然而,将这些成果转化为细胞生理条件下的成像仍然具有挑战性。改进的SRM方法的发展推动了更高的空间分辨率,当我们试图将它们用于细胞内的亚10nm荧光成像时,我们面临的问题就越大。事实证明,大多数已开发的显示纳米分辨率的SRM方法不能直接实现细胞中的分子分辨率成像。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
Methods and Applications in Fluorescence
Methods and Applications in Fluorescence CHEMISTRY, ANALYTICALCHEMISTRY, PHYSICAL&n-CHEMISTRY, PHYSICAL
CiteScore
6.20
自引率
3.10%
发文量
60
期刊介绍: Methods and Applications in Fluorescence focuses on new developments in fluorescence spectroscopy, imaging, microscopy, fluorescent probes, labels and (nano)materials. It will feature both methods and advanced (bio)applications and accepts original research articles, reviews and technical notes.
×
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