An alternative mechanism of clathrin-coated pit closure revealed by ion conductance microscopy

Shevchuk, Andrew; Novák, Pavel; Taylor, Marcus J.; Diakonov, Ivan; Ziyadeh-Isleem, Azza; Bitoun, Marc; Guicheney, Pascale; Lab, Max J.; Gorelik, Julia; Merrifield, Christien J.; Klenerman, David; Korchev, Yuri E.

doi:10.1083/jcb.201109130

Cited by 79 publications

(93 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The average cavity width and depth were 400 and 120 nm, respectively. The average cavity width was slightly larger than the size of clathrin-dependent endocytosis pits; 11 however, SICM effectively visualized nanoscale cell surface cavity clearly.…”

Section: Live Cell Imaging By Sicmmentioning

confidence: 99%

“…[5][6][7][8] Scanning ion conductance microscopy (SICM), which uses a nanopipette as a scanning probe to detect ionic current as feedback signal, was reported as an effective tool for noncontact topographical imaging of live cells, because measurements are performed under physiological conditions. [5][6][7][8][9][10][11][12][13][14] The general assembly of a SICM is shown in Fig. 1.…”

Section: Introductionmentioning

confidence: 99%

“…Previously, resolution of 3-6 nm was achieved using a 15-nmdiameter quartz nanopipette to image a membrane protein. 11 The scanning mode is a key factor when measuring convoluted live cell surface topography, because the feedback distance control system cannot predict height information in advance. Hopping mode is useful for uneven substrates over a large range of scanning areas, giving valuable information on live cell surface topography.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Nanoscale Cell Surface Topography Imaging using Scanning Ion Conductance Microscopy

et al. 2014

View full text Add to dashboard Cite

Nanoscale cell surface topography was visualized using a scanning ion conductance microscope (SICM), where a nanopipette was used as the scanning probe to detect ionic current as feedback signal. SICM was an effective tool for noncontact topographical imaging of live cells, because measurements were performed under physiological conditions. This breakthrough technique opens up a wealth of possible new experiments in membrane and cell biology.

show abstract

Section: Live Cell Imaging By Sicmmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Nanoscale Cell Surface Topography Imaging using Scanning Ion Conductance Microscopy

et al. 2014

View full text Add to dashboard Cite

show abstract

“…The use of confocal microscopy in tandem with SICM has allowed the simultaneous collection of sample topography and imaging of the spatial distribution of fluorescent molecules or particles of interest [149]. This approach has been used to study endocytic pathways [150], such as probing the mechanism of clathrin-coated pit closure [151]. Additionally fluorescence measurements have been very valuable in following the dynamics of delivery from nanopipettes and the behaviour of the fluorescent moieties while in a nanopipette [75,152].…”

Section: (D) Scanning Electrochemical Microscopy-scanning Ion Conductmentioning

confidence: 99%

Multifunctional scanning ion conductance microscopy

2017

View full text Add to dashboard Cite

Scanning ion conductance microscopy (SICM) is a nanopipette-based technique that has traditionally been used to image topography or to deliver species to an interface, particularly in a biological setting. This article highlights the recent blossoming of SICM into a technique with a much greater diversity of applications and capability that can be used either standalone, with advanced control (potential-time) functions, or in tandem with other methods. SICM can be used to elucidate functional information about interfaces, such as surface charge density or electrochemical activity (ion fluxes). Using a multi-barrel probe format, SICM-related techniques can be employed to deposit nanoscale three-dimensional structures and further functionality is realized when SICM is combined with scanning electrochemical microscopy (SECM), with simultaneous measurements from a single probe opening up considerable prospects for multifunctional imaging. SICM studies are greatly enhanced by finite-element method modelling for quantitative treatment of issues such as resolution, surface charge and (tip) geometry effects. SICM is particularly applicable to the study of living systems, notably single cells, although applications extend to materials characterization and to new methods of printing and nanofabrication. A more thorough understanding of the electrochemical principles and properties of SICM provides a foundation for significant applications of SICM in electrochemistry and interfacial science.

show abstract

“…surface of COS-7 cells revealed by scanning ion conductance microscopy. 44 Figure 8(d) shows histograms of the observed rates of pit formation for three types of cells in each of which mEGFP, mEGFP-Rab5(Q79L), or mEGFPRab5(Q79L)-CAAX is overexpressed. The nanoscale pits were infrequently observed in HeLa cells transfected only by mEGFP (1.9% of a total of 2689 frames of 27 cells), whereas the frequency of nanoscale pit appearance was significantly increased by overexpression of mEGFP-Rab5(Q79L) (19% of a total of 3994 frames of 30 cells) and further increased by overexpression of mEGFP-Rab5(Q79L)-CAAX (36% of a total of 3090 frames of 24 cells), consistent with a persistent stimulation effect of Rab5 mutants on endocytosis.…”

Section: Imaging Of Hela Cellsmentioning

confidence: 99%

Wide-area scanner for high-speed atomic force microscopy

Watanabe

Uchihashi

Kobashi

et al. 2013

Review of Scientific Instruments

View full text Add to dashboard Cite

High-speed atomic force microscopy (HS-AFM) has recently been established. The dynamic processes and structural dynamics of protein molecules in action have been successfully visualized using HS-AFM. However, its maximum scan ranges in the X-and Y-directions have been limited to ∼1 μm and ∼4 μm, respectively, making it infeasible to observe the dynamics of much larger samples, including live cells. Here, we develop a wide-area scanner with a maximum XY scan range of ∼46 × 46 μm 2 by magnifying the displacements of stack piezoelectric actuators using a leverage mechanism. Mechanical vibrations produced by fast displacement of the X-scanner are suppressed by a combination of feed-forward inverse compensation and the use of triangular scan signals with rounded vertices. As a result, the scan speed in the X-direction reaches 6.3 mm/s even for a scan size as large as ∼40 μm. The nonlinearity of the X-and Y-piezoelectric actuators' displacements that arises from their hysteresis is eliminated by polynomial-approximation-based open-loop control. The interference between the X-and Y-scanners is also eliminated by the same technique. The usefulness of this wide-area scanner is demonstrated by video imaging of dynamic processes in live bacterial and eukaryotic cells.

show abstract

An alternative mechanism of clathrin-coated pit closure revealed by ion conductance microscopy

Abstract: Simultaneous ion conductance and confocal microscopy in live cells reveal a new form of asymmetric clathrin-coated pit closure.

Cited by 79 publications

References 40 publications

Nanoscale Cell Surface Topography Imaging using Scanning Ion Conductance Microscopy

Nanoscale Cell Surface Topography Imaging using Scanning Ion Conductance Microscopy

Multifunctional scanning ion conductance microscopy

Wide-area scanner for high-speed atomic force microscopy

Contact Info

Product

Resources

About