Human Immunology Unit, Weatherall Institute of Molecular Medicine, University of Oxford, Headley Way, Oxford, OX3 9DS, UK.
Leibniz Institute of Photonic Technology e.V., Albert-Einstein Strasse 9, D-07745 Jena, Germany, Member of the Leibniz Centre for Photonics in Infection Research (LPI), Jena, Germany.
Biol Chem. 2023 Jan 26;404(2-3):87-106. doi: 10.1515/hsz-2022-0314. Print 2023 Feb 23.
Fluorescence microscopy is an important tool for studying cellular structures such as organelles. Unfortunately, many details in the corresponding images are hidden due to the resolution limit of conventional lens-based far-field microscopy. An example is the study of peroxisomes, where important processes such as molecular organization during protein important can simply not be studied with conventional far-field microscopy methods. A remedy is super-resolution fluorescence microscopy, which is nowadays a well-established technique for the investigation of inner-cellular structures but has so far to a lesser extent been applied to the study of peroxisomes. To help advancing the latter, we here give an overview over the different super-resolution microscopy approaches and their potentials and challenges in cell-biological research, including labelling issues and a focus on studies on peroxisomes. Here, we also highlight experiments beyond simple imaging such as observations of diffusion dynamics of peroxisomal proteins.
荧光显微镜是研究细胞结构(如细胞器)的重要工具。然而,由于传统基于透镜的远场显微镜的分辨率限制,许多相应图像中的细节都被隐藏了。一个例子是过氧化物酶体的研究,在过氧化物酶体中,重要的过程,如蛋白质的分子组织,根本无法用传统的远场显微镜方法进行研究。一种补救方法是超分辨率荧光显微镜,它是目前用于研究细胞内结构的一种成熟技术,但迄今为止,它在过氧化物酶体的研究中应用较少。为了帮助推进后者,我们在这里概述了不同的超分辨率显微镜方法及其在细胞生物学研究中的潜力和挑战,包括标记问题,并重点介绍了过氧化物酶体的研究。在这里,我们还强调了超越简单成像的实验,例如过氧化物酶体蛋白扩散动力学的观察。