Fluorescently Guided Optical Photothermal Infrared Microspectroscopy for Protein-Specific Bioimaging at Subcellular Level.
Fluorescently Guided Optical Photothermal Infrared Microspectroscopy for Protein-Specific Bioimaging at Subcellular Level.
复制标题
DOI：
10.1021/acs.jmedchem.2c01359
复制
发表时间：
2023-02-23
影响因子：
7.3
通讯作者：
Klementieva, Oxana
中科院分区：
文献类型：
--
作者：
Prater, Craig;Bai, Yeran;Konings, Sabine C.;Martinsson, Isak;Swaminathan, Vinay S.;Nordenfelt, Pontus;Gouras, Gunnar;Borondics, Ferenc;Klementieva, Oxana
红外光谱成像广泛用于生物分子结构的可视化，像光热红外（OPTIR）显微光谱技术等能够实现小于500纳米的空间分辨率。然而，这些方法对特定细胞类型和细胞成分缺乏特异性，因此无法作为独立技术来评估它们的特性。在此，我们开发了一种新型工具——荧光引导光热红外显微光谱技术，该技术同时利用落射荧光成像和OPTIR来进行荧光引导的红外光谱分析。这种新方法突破了红外显微镜的衍射极限，能够直接在组织和单细胞中对特定蛋白质进行结构分析。本文所述实验利用落射荧光快速定位组织或神经元培养物中的淀粉样蛋白，从而引导OPTIR测量，以在亚细胞水平评估淀粉样蛋白结构。我们认为，这种新方法将为红外光谱学增添有价值的手段，在复杂系统中进行测量时提供细胞特异性，助力对结构改变的蛋白质聚集体的研究。
Infrared spectroscopic imaging is widely used for the visualization of biomolecule structures, and techniques such as optical photothermal infrared (OPTIR) microspectroscopy can achieve <500 nm spatial resolution. However, these approaches lack specificity for particular cell types and cell components and thus cannot be used as a stand-alone technique to assess their properties. Here, we have developed a novel tool, fluorescently guided optical photothermal infrared microspectroscopy, that simultaneously exploits epifluorescence imaging and OPTIR to perform fluorescently guided IR spectroscopic analysis. This novel approach exceeds the diffraction limit of infrared microscopy and allows structural analysis of specific proteins directly in tissue and single cells. Experiments described herein used epifluorescence to rapidly locate amyloid proteins in tissues or neuronal cultures, thus guiding OPTIR measurements to assess amyloid structures at the subcellular level. We believe that this new approach will be a valuable addition to infrared spectroscopy providing cellular specificity of measurements in complex systems for studies of structurally altered protein aggregates.
登录
查看更多内容
影响因子：
2.9
作者：
Nilsson, KPR;Herland, A;Inganäs, O
通讯作者：
Inganäs, O
影响因子：
6
作者：
Freitas RO;Cernescu A;Engdahl A;Paulus A;Levandoski JE;Martinsson I;Hebisch E;Sandt C;Gouras GK;Prinz CN;Deierborg T;Borondics F;Klementieva O
通讯作者：
Klementieva O
影响因子：
5.6
作者：
Bouderlique E;Tang E;Zaworski J;Coudert A;Bazin D;Borondics F;Haymann JP;Leftheriotis G;Martin L;Daudon M;Letavernier E
通讯作者：
Letavernier E
影响因子：
16.6
作者：
Klementieva O;Willén K;Martinsson I;Israelsson B;Engdahl A;Cladera J;Uvdal P;Gouras GK
通讯作者：
Gouras GK
影响因子：
5.7
作者：
Lim, Jong Min;Park, Chanjong;Cho, Minhaeng
通讯作者：
Cho, Minhaeng