Fluorescence microscopy for visualizing single-molecule protein dynamics

• Published in: Biochimica et biophysica acta. General subjects Vol. 1864; no. 2; p. 129362
• Main Author: Yokota, Hiroaki
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.02.2020
• Subjects: fluorescence; fluorescence microscopy; fluorescent dyes; Fluorescent nanodiamond (FND); image analysis; nanodiamonds; Optically detected magnetic resonance (ODMR); photostability; Protein dynamics; Single-molecule fluorescence imaging (smFI); Super-resolution microscopy; MOPS; MPC; ZMWs; Fluorescent nanodiamond (FND); AFM; smFRET; EMCCD; NBA; TIRF; 2D; DDS; PALM; NHS; ODMR; Super-resolution microscopy; SSIM; Optically detected magnetic resonance (ODMR); FIONA; DOPI; STED; smFI; COT; Protein dynamics; Single-molecule fluorescence imaging (smFI); PCA; PCD; Qrod; STORM; 3D; PEG; FND; Qdot
• ISSN: 0304-4165; 1872-8006; 1872-8006
• DOI: 10.1016/j.bbagen.2019.05.005

Single-molecule fluorescence imaging (smFI) has evolved into a valuable method used in biophysical and biochemical studies as it can observe the real-time behavior of individual protein molecules, enabling understanding of their detailed dynamic features. smFI is also closely related to other state-of-the-art microscopic methods, optics, and nanomaterials in that smFI and these technologies have developed synergistically. This paper provides an overview of the recently developed single-molecule fluorescence microscopy methods, focusing on critical techniques employed in higher-precision measurements in vitro and fluorescent nanodiamond, an emerging promising fluorophore that will improve single-molecule fluorescence microscopy. smFI will continue to improve regarding the photostability of fluorophores and will develop via combination with other techniques based on nanofabrication, single-molecule manipulation, and so on. Quantitative, high-resolution single-molecule studies will help establish an understanding of protein dynamics and complex biomolecular systems. •Key features of single-molecule fluorescence imaging have been reviewed.•Fluorescence labeling, surface-coating methods, and modern microscopy are described.•Overview is given for frequently used fluorophores, dyes, and quantum dots.•Fluorescent nanodiamond, a promising fluorophore, is introduced.•Improved photostability and other concurrent methods can further develop the method.