Abstract:
Biological tissues are not uniquely composed of cells. A substantial part of their volume is extracellular space, which is primarily filled by an intricate network of macromolecules constituting the extracellular matrix (ECM). The ECM serves as the scaffolding for tissues and organs throughout the body, playing an essential role in their structural and functional integrity. Understanding the intimate interaction between the cells and their structural microenvironment is central to our understanding of the factors driving the formation of normal versus remodelled tissue, including the processes involved in chronic fibrotic diseases. The visualization of the ECM is a key factor to track such changes successfully. This review is focused on presenting several optical imaging microscopy modalities used to characterize different ECM components. In this review, we describe and provide examples of applications of a vast gamut of microscopy techniques, such as widefield fluorescence, total internal reflection fluorescence, laser scanning confocal microscopy, multipoint/slit confocal microscopy, two-photon excited fluorescence (TPEF), second and third harmonic generation (SHG, THG), coherent anti-Stokes Raman scattering (CARS), fluorescence lifetime imaging microscopy (FLIM), structured illumination microscopy (SIM), stimulated emission depletion microscopy (STED), ground-state depletion microscopy (GSD), and photoactivated localization microscopy (PALM/fPALM), as well as their main advantages, limitations.
摘要:
生物组织不是唯一由细胞组成的。它们体积的很大一部分是细胞外空间,其主要由构成细胞外基质(ECM)的大分子的复杂网络填充。ECM作为整个身体组织和器官的支架,在其结构和功能完整性方面发挥着重要作用。了解细胞与其结构微环境之间的密切相互作用是我们理解驱动正常组织与重塑组织形成的因素的核心,包括慢性纤维化疾病的过程。ECM的可视化是成功跟踪此类更改的关键因素。这篇评论的重点是介绍几种用于表征不同ECM成分的光学成像显微镜模式。在这次审查中,我们描述并提供了大量显微镜技术的应用示例,例如宽场荧光,全内反射荧光,激光扫描共聚焦显微镜,多点/狭缝共聚焦显微镜,双光子激发荧光(TPEF),二次和三次谐波产生(SHG,THG),相干反斯托克斯拉曼散射(CARS),荧光寿命成像显微镜(FLIM),结构化照明显微镜(SIM),受激发射耗尽显微镜(STED),基态耗尽显微镜(GSD),和光激活定位显微镜(PALM/fPALM),以及它们的主要优点,局限性。