A novel nanoscopic tool by combining AFM with STED microscopy

Harke, Benjamin; Chacko, Jenu V.; Haschke, Heiko; Canale, Claudio; Diaspro, Alberto

doi:10.1186/2192-2853-1-3

Cited by 81 publications

(81 citation statements)

References 29 publications

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“…Physicist Alberto Diaspro, director of the nanophysics department at the Italian Institute of Technology in Genoa, is correlating AFM with super-resolution stimulated emission depletion (STED) microscopy to develop tools for topographical imaging, nanomechanical imaging, and measurements of cell stiffness 3 . In STED, researchers intentionally deactivate some fluorophores in part of the sample, which enhances the resolution in that area.…”

Section: Daryl Howard Australian Synchrotronmentioning

confidence: 99%

Two microscopes are better than one

Smith¹

2012

Nature

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T here are many types of microscopy, each providing a unique set of benefits. Light microscopy of cells that express fluorescently labelled molecules, for example, lets scientists observe the movements of specific molecules or protein complexes in live cells in real time, or in fixed samples. Scanning electron microscopy (EM) reveals tiny details of the cell surface, and transmission EM shows the detailed cytoarchitecture of sections through fixed tissue. Other aspects of the structure of cells and tissues can be explored using techniques such as ion microscopy, total internal reflection microscopy, atomic force microscopy and super-resolution microscopy. Each type provides different information, but using two sorts of microscopy simultaneously provides even more -and molecular tools have been developed to link them together. Researchers testing the waters of this 'correlative microscopy' are beginning to discover its challenges and rewards.Correlated light-electron microscopy, for example, provides both the specificity and real-time observation of light microscopy with fluorescent labelling and the better structural resolution of EM. But such correlative microscopy has historically been tricky to use. Scientists who tried it might have needed to switch to EM halfway through an experiment, which meant moving the sample from one This image of a butterfly wing was constructed using two types of microscopy: a confocal image of the reflective eyespots shows scales (green) and wing (red); scanning electron microscopy reveals the different structure of non-reflective scales (upper left).

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Section: Daryl Howard Australian Synchrotronmentioning

confidence: 99%

Two microscopes are better than one

Smith¹

2012

Nature

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“…Additionally, infra-red (IR) and Raman-based imaging methods in which the spectroscopic technique has been coupled with scanning probe microscopy namely, apertureless near-field scanning infrared microscopy (ANSIM) [21,22], coherent anti-stokes Raman scattering (CARS) [23] and tip-enhanced Raman scattering (TERS) [24][25][26][27] have also been utilized to extract both topographical and molecular structural details of a variety of biological systems. Recently a "nanoscopic tool", comprising a combination of AFM and STED microscopy, for imaging biological samples has been reported [28]. In the current review, we discuss a few recent applications of Unauthenticated Download Date | 5/9/18 9:02 PM nanophotonics in unravelling the structural details of an important class of biological supramolecular assemblies, namely amyloids that are made up of proteins and are the hallmarks of a variety of neurodegenerative disorders.…”

Section: Near-field Scanning Optical Microscopy (Nsom): Principlesmentioning

confidence: 99%

Nanophotonics of protein amyloids

Bhattacharya¹,

Mukhopadhyay²

2014

Nanophotonics

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Technological breakthroughs in the superresolution optical imaging techniques have enriched our current understanding of a range of biological systems and biomolecular processes at the nanoscopic spatial resolution. Protein amyloids are an important class of ordered protein assemblies consisting of misfolded proteins that are implicated in a wide range of devastating human diseases. In order to decipher the structural basis of the supramolecular protein assembly in amyloids and their detrimental interactions with the cell membranes, it is important to employ high-resolution optical imaging techniques. Additionally, amyloids could serve as novel biological nanomaterials for a variety of potential applications. In this review, we summarize a few examples of the utility of near-field scanning optical imaging methodologies to obtain a wealth of structural information into the nanoscale amyloid assembly. Although the near-field technologies were developed several decades ago, it is only recently that these methodologies are being applied and adapted for amyloid research to yield novel information pertaining to the exciting nanoscopic world of protein aggregates. We believe that the account on the nanophotonics of amyloids described in this review will be useful for the future studies on the biophysics of amyloids.

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“…For example, STED microscopy has been coupled to AFM (so named STED-AFM) to image ATTO-647-immunolabeled microtubules in fixed HeLa and kidney cos7 cells. 95,96 STED-AFM offers high-resolution fluorescent images with molecular specificity, topological images of the sample, and information about the mechanical properties, such as elasticity and local stiffness of microtubule filaments, in the same sample space. 95,96 In another approach, STED microscopy has been coupled with optical tweezers to integrate both nanoscale resolution and precise mechanical control of the sample.…”

Section: Concluding Remarks and Future Perspectivesmentioning

confidence: 99%

Review of recent developments in stimulated emission depletion microscopy: applications on cell imaging

2014

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Abstract. Stimulated emission depletion (STED) microscopy is one type of far-field optical technique demonstrated to provide subdiffraction resolution. STED microscopy utilizes a donut-shaped depletion beam to limit the probe volume to be much smaller than a diffraction-limited spot. Resolutions as small as a few tens of nanometers laterally are reported for cell analysis. The different versions of STED microscopes are described and contrasted in terms of their applicability for biological imaging. Finally, we suggest likely avenues for improving the performance and increasing the utility of STED microscopy.

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A novel nanoscopic tool by combining AFM with STED microscopy

Cited by 81 publications

References 29 publications

Two microscopes are better than one

Two microscopes are better than one

Nanophotonics of protein amyloids

Review of recent developments in stimulated emission depletion microscopy: applications on cell imaging

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