Hongjun Wu, Yalan Zhao, Xiao Zhou, Tianxiao Wu, Jiaming Qian, Shijia Wu, Yongtao Liu, Chao Zuo, "Super-resolution microscopy reveals new insights into organelle interactions," Adv. Imaging 1, 032001 (2024)
- Advanced Imaging
- Vol. 1, Issue 3, 032001 (2024)
![The basic principle and application of SMLM. (a) The basic principle of STORM[9]. (b) Optical path and spectral modulation process of traditional STORM. (c) 4Pi-STORM imaging of the neuronal cytoskeleton[11]. (d) The process of image reconstruction based on a deep learning network. (e) Comparison of basal membrane profiles of three different cell types[25]. (f) STORM (left) and wide-field (right) images of microtubules in HepG2 cells labeled with 565 quantum dots (QDs)[10]. (g) 4Pi-STORM imaging of mitochondrial crista in the U-2 OS cell and COS-7 cell, respectively[11].](/richHtml/ai/2024/1/3/032001/img_001.png)
Fig. 1. The basic principle and application of SMLM. (a) The basic principle of STORM[9]. (b) Optical path and spectral modulation process of traditional STORM. (c) 4Pi-STORM imaging of the neuronal cytoskeleton[11]. (d) The process of image reconstruction based on a deep learning network. (e) Comparison of basal membrane profiles of three different cell types[25]. (f) STORM (left) and wide-field (right) images of microtubules in HepG2 cells labeled with 565 quantum dots (QDs)[10]. (g) 4Pi-STORM imaging of mitochondrial crista in the U-2 OS cell and COS-7 cell, respectively[11].
![The basic principle and application of STED. (a) The basic principle of STED[12]. (b) Optical path and spectral modulation process of two-color STED[27]. (c) Diffusion of Atto647N-DPPE in the membrane of an XTC cell in STED[13]. (d) Immunolabeled Munc13-1 and RIM1 molecules were visualized by STED microscopy at the hMFBs[43]. (e) Diagnostics and understanding disease mechanisms in STED and confocal microscopy[29]. (f) The imaging of microtubules in fixed BSC-1 cells in confocal microscopy, STED, and FM-STED[39].](/richHtml/ai/2024/1/3/032001/img_002.png)
Fig. 2. The basic principle and application of STED. (a) The basic principle of STED[12]. (b) Optical path and spectral modulation process of two-color STED[27]. (c) Diffusion of Atto647N-DPPE in the membrane of an XTC cell in STED[13]. (d) Immunolabeled Munc13-1 and RIM1 molecules were visualized by STED microscopy at the hMFBs[43]. (e) Diagnostics and understanding disease mechanisms in STED and confocal microscopy[29]. (f) The imaging of microtubules in fixed BSC-1 cells in confocal microscopy, STED, and FM-STED[39].
![The basic principle and application of SIM. (a) Optical path and spectral modulation process of traditional SIM (linear). (b) Spectral modulation process of SSIM. (c) F-actin protein of COS-7 cells imaged by SSIM and other methods (top left: wide field, top right: deconvolution, bottom left: SIM, bottom right: SSIM)[14]. (d) The structure of mesoplasmic membrane microcapsules in COS-7 cells from different methods (top left: wide field, top right: deconvolution, bottom left: SIM, bottom right: SSIM)[14]. (e) The SSIM image of mesoplasmic membrane microcapsules in live COS-7 cells[14]. (f) The basic principle of 3D-SIM[21]. (g) Imaging results of traditional widefield and 3D-SIM in live HeLa cells[49].](/Images/icon/loading.gif){kind=icon}
Fig. 3. The basic principle and application of SIM. (a) Optical path and spectral modulation process of traditional SIM (linear). (b) Spectral modulation process of SSIM. (c) F-actin protein of COS-7 cells imaged by SSIM and other methods (top left: wide field, top right: deconvolution, bottom left: SIM, bottom right: SSIM)[14]. (d) The structure of mesoplasmic membrane microcapsules in COS-7 cells from different methods (top left: wide field, top right: deconvolution, bottom left: SIM, bottom right: SSIM)[14]. (e) The SSIM image of mesoplasmic membrane microcapsules in live COS-7 cells[14]. (f) The basic principle of 3D-SIM[21]. (g) Imaging results of traditional widefield and 3D-SIM in live HeLa cells[49].
Fig. 4. The basic principle and application of MINFLUX. (a) The basic principle of MINFLUX[16]. (b) Optical path and spectral modulation process of 3D MINFLUX[17]. (c) Arrangement of up to nine on-off switchable fluorophores on the origami (top) and the smaller DNA origami structure (bottom)[16]. (d) Single-molecule MINFLUX imaging and precise tracking in live E. coli bacteria[16]. (e) 3D MINFLUX raw data and corresponding information at different dimensions (top) and clathrin visualized by SNAP labeling in HeLa cells (bottom)[17]. (f) Distribution of Mic60 in mitochondria of human U-2OS cells[18]. (g) Two-color 3D MINFLUX acquisition of a mitochondrion in human dermal fibroblasts[18].
Fig. 5. The properties and interaction of mitochondria and lysosomes. (a) Comparative summed-molecule TIRF, PALM, TEM, and PALM+TEM overlay images of mitochondria in a cryo-prepared thin section from a COS-7 cell expressing dEosFP-tagged cytochrome-C oxidase import sequence[87]. (b) SIM image of lysosome in HeLa cell magnified images from the dashed box at different time points[94]. (c) The movement status of mitochondria at different time points (left) and the whole tubulation retraction process (right) were imaged by STORM[90]. (d) The fusion of two mitochondria[51]. (e) The fission of one mitochondrion (bottom) was imaged by SIM[51].(f) TRPML1 activation preferentially increases mitochondrial calcium at mitochondria (red)-lysosome green contacts[101]. (g) The behavior of autophagy after the treatment of the antimicrobial drug (autophagy marker protein LC3 and p63)[97]. (h) Dual-color SIM images of dynamic physical interactions between lysosomes and mitochondria in live U2OS cells (Atto 647 N-magenta, Lysosome-565-green[99]. (i) Determination of co-localization of mitochondria with lysosomes after serum starvation[99].
Fig. 6. The properties and interaction of mitochondria and endoplasmic reticulum. (a) The diameter of ER tubules labeled with SNAP-Sec61β was imaged. The spatial distribution of endoplasmic reticulum changes with the fluidity of live cells by STED[104]. (b) ER distribution changes during autophagy in U2OS cells[107]. (c) The morphological change of mitochondria during the process of autophagy[109]. (d) The morphological change of ER in live Hela cells was monitored by dSTORM under stress conditions[105]. (e) MDT at ER (magenta)–mitochondria (green) contacts were imaged by SIM[89]. (f) Two-color STED time-lapse imaging of ER-mitochondria interaction within a neurite[108].
Fig. 7. The properties and interaction of the mitochondria and nucleus. (a) Representative confocal images of mito-STAR protein in HeLa and INS-1 cells[118]. (b) Two-color 4Pi microscopy images of PML and SUMO proteins[116]. (c) 3D tomography of interaction sites for Imp
Fig. 8. The properties and interaction of the endoplasmic reticulum and Golgi apparatus. (a) Confocal and STED images of the Golgi[121]. (b) Complex stacked architecture of the Golgi apparatus by the 4Pi single-molecule switched super-resolution microscope[91]. (c) The distribution of MICAL-L1/TGN46, MICAL-L1/ GM130, and MICAL-L1/TfR colocalization was evaluated by STORM[125]. (d) Cargo (green) is transported from cis-Golgi (red) to TGN (blue) while being maintained in a maturing cisterna[122]. (e) 4D images of the Golgi apparatus, TGN, and secretory trafficking zone component clathrin in the epidermal cell of the root elongation zone under SCLIM[125].
Fig. 9. The properties and interaction of membraneless organelles. (a) Super-resolution 3D-SIM imaging of nucleolar organization[134]. (b) The super-resolution imaging of centriole distal by dual-channel dSTORM[128]. (c) The biological communication of the cytoplasm by the NPC[132]. (d) SIM-TIRF time-lapse of the self-assembling process of the centrosome in vitro [127]. (e) 3D and 2D dSTORM images of U-ExM expanded and re-embedded chlamydomonas centrioles[127].
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Table 1. System Parameters of Different Techniques.
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