A super-resolution platform for correlative live single-molecule imaging and STED microscopy

Inavalli, V. V. G. Krishna; Lenz, Martin O.; Butler, Corey; Angibaud, Julie; Compans, Benjamin; Levet, Florian; Tønnesen, Jan; Rossier, Olivier; Giannone, Grégory; Thoumine, Olivier; Hosy, Eric; Choquet, Daniel; Sibarita, Jean‐Baptiste; Nägerl, U. Valentin

doi:10.1038/s41592-019-0611-8

Cited by 56 publications

(50 citation statements)

References 26 publications

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“…However, this is still a significant challenge for the single molecule localization microscopy (Orré et al, 2019) such as PALM (Betzig et al, 2006), stochastic optical reconstruction microscopy (Rust et al, 2006) or super-resolution optical fluctuation imaging (Dertinger et al, 2009). First promising insights into the dynamics of NAs nevertheless were provided by the single particle tracking PALM (Inavalli et al, 2019), which revealed integrin cycling between free diffusion and immobilization, while transient interaction with talins promoted integrin activation and immobilization (Rossier et al, 2012). Further studies of integrin dynamics will require techniques such that can operate at micro second-time scales with 1 nm precision of molecules located few nanometers apart.…”

Section: Challenges and Perspectives In The Field Of Nascent Adhesionsmentioning

confidence: 99%

Recent Advances and Prospects in the Research of Nascent Adhesions

et al. 2020

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Nascent adhesions are submicron transient structures promoting the early adhesion of cells to the extracellular matrix. Nascent adhesions typically consist of several tens of integrins, and serve as platforms for the recruitment and activation of proteins to build mature focal adhesions. They are also associated with early stage signaling and the mechanoresponse. Despite their crucial role in sampling the local extracellular matrix, very little is known about the mechanism of their formation. Consequently, there is a strong scientific activity focused on elucidating the physical and biochemical foundation of their development and function. Precisely the results of this effort will be summarized in this article.

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Section: Challenges and Perspectives In The Field Of Nascent Adhesionsmentioning

confidence: 99%

Recent Advances and Prospects in the Research of Nascent Adhesions

et al. 2020

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“…It is also compatible with thick samples and standard fluorophores, but cannot be used for live imaging. Also noteworthy is the combined use of PALM and uPAINT with 3D-STED or SUSHI imaging methods onto a single microscope platform (Inavalli et al, 2019). By combining the strengths of deterministic and stochastic SRM methods, this system allows the simultaneous investigation of both the nanoscale morphology of synapses and the localization and dynamics of synaptic proteins in living samples and opens the possibility of applying a similar approach using other SRM techniques.…”

Section: Perspectives For Srm Of the Presynaptic Compartmentmentioning

confidence: 99%

Unraveling the Nanoscopic Organization and Function of Central Mammalian Presynapses With Super-Resolution Microscopy

et al. 2021

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The complex, nanoscopic scale of neuronal function, taking place at dendritic spines, axon terminals, and other minuscule structures, cannot be adequately resolved using standard, diffraction-limited imaging techniques. The last couple of decades saw a rapid evolution of imaging methods that overcome the diffraction limit imposed by Abbe’s principle. These techniques, including structured illumination microscopy (SIM), stimulated emission depletion (STED), photo-activated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM), among others, have revolutionized our understanding of synapse biology. By exploiting the stochastic nature of fluorophore light/dark states or non-linearities in the interaction of fluorophores with light, by using modified illumination strategies that limit the excitation area, these methods can achieve spatial resolutions down to just a few tens of nm or less. Here, we review how these advanced imaging techniques have contributed to unprecedented insight into the nanoscopic organization and function of mammalian neuronal presynapses, revealing new organizational principles or lending support to existing views, while raising many important new questions. We further discuss recent technical refinements and newly developed tools that will continue to expand our ability to delve deeper into how synaptic function is orchestrated at the nanoscopic level.

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“…STORM imaging enabled the discovery of aligned nanocolumns of clustered presynaptic release machinery and postsynaptic receptors apposed across the synaptic cleft, an arrangement that promotes efficient neurotransmission ( Figure 5K; Tang et al, 2016). These subsynaptic domains (SSDs) have been further investigated using STED (Nair et al, 2013;Broadhead et al, 2016;Dzyubenko et al, 2016;Hruska et al, 2018;Masch et al, 2018;Wegner et al, 2018;Wiesner et al, 2020), SMLM (MacGillavry et al, 2013Nair et al, 2013;Specht et al, 2013;Broadhead et al, 2016;Pennacchietti et al, 2017;Sinnen et al, 2017;Kellermayer et al, 2018;Inavalli et al, 2019;Ferreira et al, 2020;Goncalves et al, 2020;Yang et al, 2020), PAINT (Nair et al, 2013), and SIM (Crosby et al, 2019). Subsynaptic clusters of glutamatergic receptors are developmentally-regulated (Kellermayer et al, 2018) and SSDs containing receptors and critical synaptic machinery show activity-dependent changes in response to stimulation protocols for inducing synaptic potentiation, depression, and homeostatic plasticity (MacGillavry et al, 2013;Tang et al, 2016;Pennacchietti et al, 2017;Sinnen et al, 2017;Ferreira et al, 2020;Wiesner et al, 2020;Yang et al, 2020).…”

Section: Super-resolution Imaging Of Neural Development and Plasticitmentioning

confidence: 99%

A Picture Worth a Thousand Molecules—Integrative Technologies for Mapping Subcellular Molecular Organization and Plasticity in Developing Circuits

Minehart

Speer

2021

Front. Synaptic Neurosci.

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A key challenge in developmental neuroscience is identifying the local regulatory mechanisms that control neurite and synaptic refinement over large brain volumes. Innovative molecular techniques and high-resolution imaging tools are beginning to reshape our view of how local protein translation in subcellular compartments drives axonal, dendritic, and synaptic development and plasticity. Here we review recent progress in three areas of neurite and synaptic study in situ—compartment-specific transcriptomics/translatomics, targeted proteomics, and super-resolution imaging analysis of synaptic organization and development. We discuss synergies between sequencing and imaging techniques for the discovery and validation of local molecular signaling mechanisms regulating synaptic development, plasticity, and maintenance in circuits.

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A super-resolution platform for correlative live single-molecule imaging and STED microscopy

Cited by 56 publications

References 26 publications

Recent Advances and Prospects in the Research of Nascent Adhesions

Recent Advances and Prospects in the Research of Nascent Adhesions

Unraveling the Nanoscopic Organization and Function of Central Mammalian Presynapses With Super-Resolution Microscopy

A Picture Worth a Thousand Molecules—Integrative Technologies for Mapping Subcellular Molecular Organization and Plasticity in Developing Circuits

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