Infrared and Laser Engineering, Volume. 51, Issue 11, 20220622(2022)
Application of super-resolution microscopy in the study of organelle interactions (invited)
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![Principle of stimulated emission depletion (STED) microscopy[20]](/Images/icon/loading.gif){kind=icon}
Fig. 1. Principle of stimulated emission depletion (STED) microscopy[20]
Fig. 3. Principle of single molecule localization microscopy (SMLM)[31]
Fig. 4. SIM images of mitochondria (green) and lysosomes (red) contact in wild type living cells[38]
Fig. 5. Mitochondria distribution guided by cytoskeleton during mitosis imaged by GI-SIM[48]（Red：Cytoskeleton； Blue：Mitochondria）
Fig. 6. STED showed the mitochondrion-lipid droplet interface. (a) Expression of PLIN5 (red) at interface of mitochondria and lipid droplet; (b) Voxelized rendering of extracted LD, mitochondria and overlapping zones between them (LD (green), mitochondria (red) and computed contact zones (blue))[55-56]
Fig. 7. Time-lapse images of a typical mitochondrial fission and fusion event at an ER-Mito contact site captured by GI-SIM[64]. (a)-(b) Mitochondrial fission mediated by ER; (c)-(d) Mitochondrial fusion mediated by ER
Fig. 8. SIM images showed that SEPN1-devoid cells display less ER-mitochondria contact[68] （WT: Wild type cells; SEPN1 KO: SEPN1 knock out cells）
Table 1. Principles and characteristics of three types of super-resolution microscopy
View tableView in ArticleTable 1. Principles and characteristics of three types of super-resolution microscopy
Principle xy resolution/nm z resolution/nm Fluorescent molecules Live cell imaging Image reconstruction STED Stimulated emission depletion based on PSF ~40 ~70 Free Yes Not required SIM Structured illumination ~50 ~250 Free Yes Required PALM Photo activated localization based on photoswitchable fluorescent molecules ~20 ~50 Photoswitchable Yes Required STORM Stochastic optical reconstruction based on photoswitchable fluorescent molecules ~20 ~50 Photoswitchable Yes Required
Table 2. Application of super-resolution microscopy in the study of organelle interactions
View tableView in ArticleTable 2. Application of super-resolution microscopy in the study of organelle interactions
Organelle interactions Research contents Super-resolution microscopy Cells Mitochondria-lysosome Mitochondria and lysosome contact ( MLC) marks sites of mitochondrial fission and the formation and release of MLC is bidirectionally regulated by mitochondrial and lysosomal dynamic[ 36 ]; Endoplasmic reticulum (ER) recruits lysosomes to act in concert at the fission site for the efficient division of mitochondria[37 ]; The analysis methods of mitochondria-lysosome interactions based on structured illumination microscopy(SIM)[ 38 ]; Lysosome-targeted biosensor for the super-resolution imaging of lysosome-mitochondrion interaction based on SIM[39 ].SIM HT22 cells; SHSY-5Y cells; HeLa cells; HSF cells Mitochondria- nucleus Observation of mitochondria-nucleus contact and the change of expression sites of Sirt4 from mitochondria to nucleus under mitochondrial stress conditions[ 43 -44 ].SIM HeLa cells; Pancreaticβcells Mitochondria-cytoskeleton Mitochondrial morphology and distribution are regulated by cytoskeleton. During mitosis, dense meshwork of subcortical actin cables organizes three-dimensional mitochondrial positioning to ensure both equal and random inheritance of mitochondria in symmetrically dividing cells[ 48 ]; Actin maintains microtubule organization, dynamics and stability by affecting tubulin acetylation levels and further regulate mitochondrial distribution[49 ]; Cytoskeleton regulates fission and fusion of mitochondria[50 ].SIM; STED Hela cells; COS-7 cells; HEK293 cells; U2 OS cells Mitochondria-lipid droplet Observation of mitochondria-lipid droplets contacts; Overexpression of perilipin5 leads to increased number of mitochondria surrounding lipid droplets[ 55 ].STED COS-7 cells Mitochondria-Endoplasmic reticulum Tubular endoplasmic reticulum regulates mitochondrial fission and fusion[ 64 ]; Calcium transients on the sites of mitochondria-endoplasmic reticulum contacts[67 ]; In SEPN1-related myopathy, SEPN1 deficiency results in less mitochondria-endoplasmic reticulum contacts, calcium contents and damaged oxidative phosphorylation process[68 ].SIM COS-7 cells; U2 OS cells; HeLa cells Endoplasmic reticulum-lysosome Lysosomes moved synchronously with local endoplasmic reticulum. The anchorage of lysosomes to endoplasmic reticulum growth tips is critical for endoplasmic reticulum tubule elongation and connection[ 15 ]; Endoplasmic reticulum contacts with the edge of lysosome, which promotes the long-distance transportation of lysosome[64 ].SIM COS-7 cells; U2 OS cells Endoplasmic reticulum-cytoskeleton Endoplasmic reticulum anchors to microtubules, which guides the formation of new endoplasmic reticulum tubule branches[ 64 ]; Endoplasmic reticulum dynamics play important roles in microtubules distribution[15 ].SIM; STED COS-7 cells; U2 OS cells
Table 3. Application parameters of super-resolution microscopy
View tableView in ArticleTable 3. Application parameters of super-resolution microscopy
STED SIM SMLM xy resolution/nm ~40 ~50 ~20 z resolution/nm ~70 ~250 ~50 Temporal resolution 5 ms-2 s 10-500 ms 1 min-1 h Light intensity Medium-high Low-medium Low-medium Live cell dynamic imaging Medium Good Poor
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Taiqiang Dai, Ye Gao, Ying Ma, Bolei Cai, Fuwei Liu, Boling He, Jie Yu, Yan Hou, Peng Gao, Liang Kong. Application of super-resolution microscopy in the study of organelle interactions (invited)[J]. Infrared and Laser Engineering, 2022, 51(11): 20220622
Paper Information
Category: Special issue-Fluorescence microscopy: techniques and applications
Received: Aug. 31, 2022
Accepted: --
Published Online: Feb. 9, 2023
The Author Email: Kong Liang (liangkong2014@163.com)
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