Super-resolution imaging of fluorescent dipoles via polarized structured illumination microscopy

Zhanghao, Karl; Chen, Xingye; Liu, Wenhui; Li, Meiqi; Liu, Y.; Wang, Yiming; Luo, Sha; Xiao, Wang; Shan, Chunyan; Xie, Hao; Gao, Juntao; Chen, Xiaowei; Jin, Dayong; Li, Xiangdong; Zhang, Yan; Dai, Qionghai; Xi, Peng

doi:10.1038/s41467-019-12681-w

Cited by 109 publications

(96 citation statements)

References 46 publications

(58 reference statements)

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“…2f, g). Nevertheless, SIM improves only the spatial imaging and hyperspectral detection, but not the polarization imaging due to the missing cross harmonics, based on our previous work of polarized SIM 13 .…”

Section: Resultsmentioning

confidence: 99%

High-dimensional super-resolution imaging reveals heterogeneity and dynamics of subcellular lipid membranes

Zhanghao

Liu

et al. 2020

Nat Commun

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Lipid membranes are found in most intracellular organelles, and their heterogeneities play an essential role in regulating the organelles’ biochemical functionalities. Here we report a Spectrum and Polarization Optical Tomography (SPOT) technique to study the subcellular lipidomics in live cells. Simply using one dye that universally stains the lipid membranes, SPOT can simultaneously resolve the membrane morphology, polarity, and phase from the three optical-dimensions of intensity, spectrum, and polarization, respectively. These high-throughput optical properties reveal lipid heterogeneities of ten subcellular compartments, at different developmental stages, and even within the same organelle. Furthermore, we obtain real-time monitoring of the multi-organelle interactive activities of cell division and successfully reveal their sophisticated lipid dynamics during the plasma membrane separation, tunneling nanotubules formation, and mitochondrial cristae dissociation. This work suggests research frontiers in correlating single-cell super-resolution lipidomics with multiplexed imaging of organelle interactome.

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Section: Resultsmentioning

confidence: 99%

High-dimensional super-resolution imaging reveals heterogeneity and dynamics of subcellular lipid membranes

Zhanghao

Liu

et al. 2020

Nat Commun

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“…To overcome this problem, super-resolution fluorescence microscopy (SRM) offers the possibility to increase the high temporal and spatial resolution to better appreciate the dynamics and fine structure of ER-mitochondria contact sites [92,93]. Structured illumination microscopy (SIM), a type of SRM, is suitable for fast live-cell imaging and has been used to reveal subcellular structures and dynamics of ER-mitochondria contacts [94,95]. However, the use of this kind of approach is still limited in the characterization of ER-mitochondria contact sites as it requires highly dedicated microscopes and technical expertise, thus resulting in an expensive and difficult technique.…”

Section: Fluorescent Probe-based Methodsmentioning

confidence: 99%

ER–Mitochondria Contact Sites Reporters: Strengths and Weaknesses of the Available Approaches

Giamogante

Barazzuol

Brini

et al. 2020

IJMS

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Organelle intercommunication represents a wide area of interest. Over the last few decades, increasing evidence has highlighted the importance of organelle contact sites in many biological processes including Ca2+ signaling, lipid biosynthesis, apoptosis, and autophagy but also their involvement in pathological conditions. ER–mitochondria tethering is one of the most investigated inter-organelle communications and it is differently modulated in response to several cellular conditions including, but not limited to, starvation, Endoplasmic Reticulum (ER) stress, and mitochondrial shape modifications. Despite many studies aiming to understand their functions and how they are perturbed under different conditions, approaches to assess organelle proximity are still limited. Indeed, better visualization and characterization of contact sites remain a fascinating challenge. The aim of this review is to summarize strengths and weaknesses of the available methods to detect and quantify contact sites, with a main focus on ER–mitochondria tethering.

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“…Several SRMs, such as stimulated emission depletion microscopy (STEDM), photoactivation localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM), have been developed for imaging of nanoscale structural details and dynamics of MCSs, such as ER-PM MCS (Hsieh et al, 2017;Nascimbeni et al, 2017), ER-mitochondria MCS (Shim et al, 2012;Modi et al, 2019), ER-lysosome MCS (Shim et al, 2012), mitochondria-peroxisome MCS (Galiani et al, 2016). Among these SRMs, SIM is suitable for fast live-cell imaging and has been used to reveal numerous subcellular structures and dynamics (Li et al, 2015;Nixon-Abell et al, 2016;Zhanghao et al, 2019). Recently, ER-PM MCS in U2OS, Jurkat T and HEK293 cells was observed by SIM with total internal reflection fluorescence microscopy (TIRF-SIM) (Guo M. et al, 2018;Kang et al, 2019).…”

Section: Visualization Of Mcss By Em and Srmmentioning

confidence: 99%

Current and Emerging Approaches for Studying Inter-Organelle Membrane Contact Sites

Huang

Jiang

et al. 2020

Front. Cell Dev. Biol.

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Inter-organelle membrane contact sites (MCSs) are classically defined as areas of close proximity between heterologous membranes and established by specific proteins (termed tethers). The interest on MCSs has rapidly increased in the last years, since MCSs play a crucial role in the transfer of cellular components between different organelles and have been involved in important cellular functions such as apoptosis, organelle division and biogenesis, and cell growth. Recently, an unprecedented depth and breadth in insights into the details of MCSs have been uncovered. On one hand, extensive MCSs (organelles interactome) are revealed by comprehensive analysis of organelle network with high temporal-spatial resolution at the system level. On the other hand, more and more tethers involving in MCSs are identified and further works are focusing on addressing the role of these tethers in regulating the function of MCSs at the molecular level. These enormous progresses largely depend on the powerful approaches, including several different types of microscopies and various biochemical techniques. These approaches have greatly accelerated recent advances in MCSs at the system and molecular level. In this review, we summarize the current and emerging approaches for studying MCSs, such as various microscopies, proximity-driven fluorescent signal generation and proximity-dependent biotinylation. In addition, we highlight the advantages and disadvantages of the techniques to provide a general guidance for the study of MCSs.

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Super-resolution imaging of fluorescent dipoles via polarized structured illumination microscopy

Cited by 109 publications

References 46 publications

High-dimensional super-resolution imaging reveals heterogeneity and dynamics of subcellular lipid membranes

High-dimensional super-resolution imaging reveals heterogeneity and dynamics of subcellular lipid membranes

ER–Mitochondria Contact Sites Reporters: Strengths and Weaknesses of the Available Approaches

Current and Emerging Approaches for Studying Inter-Organelle Membrane Contact Sites

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