SRRF: Universal live-cell super-resolution microscopy

Culley, Siân; Tosheva, Kalina L.; Pereira, Pedro M.; Henriques, Ricardo

doi:10.1016/j.biocel.2018.05.014

Cited by 148 publications

(151 citation statements)

References 30 publications

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“…New improvements in superresolution (Culley, Tosheva, Matos Pereira, & Henriques, 2018;Wang, Taki, et al, 2019) and correlative microscopies (Vassilopoulos, Gibaud, Jimenez, Caillol, & Leterrier, 2019) should soon enable us to capture the dynamic itineraries of ankyrins and spectrins at nanoscale resolution, which will in turn help us more precisely pinpoint their roles in, and individual commitments to, known and novel pathways. This will represent a significant step in deciphering the code behind their modes of functional partitioning and long-range and local organization.…”

Section: Perspectivesmentioning

confidence: 99%

Cargo hold and delivery: Ankyrins, spectrins, and their functional patterning of neurons

Lorenzo

2020

Cytoskeleton

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The highly polarized, typically very long, and nonmitotic nature of neurons present them with unique challenges in the maintenance of their homeostasis. This architectural complexity serves a rich and tightly controlled set of functions that enables their fast communication with neighboring cells and endows them with exquisite plasticity. The submembrane neuronal cytoskeleton occupies a pivotal position in orchestrating the structural patterning that determines local and long‐range subcellular specialization, membrane dynamics, and a wide range of signaling events. At its center is the partnership between ankyrins and spectrins, which self‐assemble with both remarkable long‐range regularity and micro‐ and nanoscale specificity to precisely position and stabilize cell adhesion molecules, membrane transporters, ion channels, and other cytoskeletal proteins. To accomplish these generally conserved, but often functionally divergent and spatially diverse, roles these partners use a combinatorial program of a couple of dozens interacting family members, whose code is not fully unraveled. In a departure from their scaffolding roles, ankyrins and spectrins also enable the delivery of material to the plasma membrane by facilitating intracellular transport. Thus, it is unsurprising that deficits in ankyrins and spectrins underlie several neurodevelopmental, neurodegenerative, and psychiatric disorders. Here, I summarize key aspects of the biology of spectrins and ankyrins in the mammalian neuron and provide a snapshot of the latest advances in decoding their roles in the nervous system.

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Section: Perspectivesmentioning

confidence: 99%

Cargo hold and delivery: Ankyrins, spectrins, and their functional patterning of neurons

Lorenzo

2020

Cytoskeleton

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“…There have been initial reports of TPE fluoresence in nanodiamonds (29)(30)(31), showing that they are compatible D R A F T with this mode of imaging. In this paper we further demonstrate TPE imaging of NV centres in ND and show how ND is particularly suited to computational super-resolution techniques, such as that enabled by the super-resolution radial fluctuations (SRRF) (32) approach. While multi-photon microscopy is inherently suited to imaging in optically aberrating samples, it nevertheless benefits from adaptive optical image correction (33)(34)(35).…”

Section: Introductionmentioning

confidence: 86%

Nanodiamonds enable adaptive-optics enhanced, super-resolution, two-photon excitation microscopy

Johnstone

Cairns

Patton

2019

Preprint

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Particles of diamond in the 5 to 100 nm size range, known as nanodiamond, have shown promise as robust fluorophores for optical imaging. We demonstrate here that, due to their photostability, they are not only suitable for two-photon imaging, but they allow significant resolution enhancement when combined with computational super-resolution techniques. We observe a resolution of 42.5 nm when processing two-photon images with the Super-Resolution Radial Fluctuations algorithm. We show manipulation of the point-spread function of the microscope using adaptive optics. This demonstrates how the photostability of nanodiamond can also be of use when characterising adaptive optics technologies or testing the resilience of super-resolution or aberration correction algorithms. Nanodiamond | Super-resolution | Multi-photon Excitation | Adaptive Optics Correspondence: brian.patton@strath.ac.uk Patton et al. | bioRχiv | March 22, 2019 | 1-7

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“…The shape of the whole cell was manually outlined by a person blind to experimental conditions and localizations outside this shape were not used for analysis. The correction of drift between individual channels was performed using NanoJ‐Core Fiji plugin . Diffusion coefficients for individual tracks were calculated by analyzing the results of the Trackmate plugin with the MATLAB‐based @msdanalyzer software .…”

Section: Methodsmentioning

confidence: 99%

Components of the G_s signaling cascade exhibit distinct changes in mobility and membrane domain localization upon β₂‐adrenergic receptor activation

Bondar

Jang

Sviridova

et al. 2020

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The G protein signaling cascade is a key player in cell signaling. Cascade activation leads to a redistribution of its members in various cellular compartments. These changes are likely related to the “second wave” of signaling from endosomes. Here, we set out to determine whether Gs signaling cascade members expressed at very low levels exhibit altered mobility and localize in clathrin‐coated structures (CCSs) or caveolae upon activation by β2‐adrenergic receptors (β2AR). Activated β2AR showed decreased mobility and sustained accumulation in CCSs but not in caveolae. Arrestin 3 translocated to the plasma membrane after β2AR activation and showed very low mobility and pronounced accumulation in CCSs. In contrast, Gαs and Gγ2 exhibited a modest reduction in mobility but no detectable accumulation in or exclusion from CCSs or caveolae. The effector adenylyl cyclase 5 (AC5) showed a slight mobility increase upon β2AR stimulation, no redistribution to CCSs, and weak activation‐independent accumulation in caveolae. Our findings show an overall decrease in the mobility of most activated Gs signaling cascade members and confirm that β2AR and arrestin 3 accumulate in CCSs, while Gαs, Gγ2 and AC5 can transiently enter CCSs and caveolae but do not accumulate in and are not excluded from these domains.

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SRRF: Universal live-cell super-resolution microscopy

Cited by 148 publications

References 30 publications

Cargo hold and delivery: Ankyrins, spectrins, and their functional patterning of neurons

Cargo hold and delivery: Ankyrins, spectrins, and their functional patterning of neurons

Nanodiamonds enable adaptive-optics enhanced, super-resolution, two-photon excitation microscopy

Components of the G_s signaling cascade exhibit distinct changes in mobility and membrane domain localization upon β₂‐adrenergic receptor activation

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SRRF: Universal live-cell super-resolution microscopy

Cited by 148 publications

References 30 publications

Cargo hold and delivery: Ankyrins, spectrins, and their functional patterning of neurons

Cargo hold and delivery: Ankyrins, spectrins, and their functional patterning of neurons

Nanodiamonds enable adaptive-optics enhanced, super-resolution, two-photon excitation microscopy

Components of the Gs signaling cascade exhibit distinct changes in mobility and membrane domain localization upon β2‐adrenergic receptor activation

Contact Info

Product

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Components of the G_s signaling cascade exhibit distinct changes in mobility and membrane domain localization upon β₂‐adrenergic receptor activation