Super-Resolution Microscopy: From Single Molecules to Supramolecular Assemblies

Sydor, Andrew M.; Czymmek, Kirk J.; Puchner, Elias M.; Mennella, Vito

doi:10.1016/j.tcb.2015.10.004

Cited by 236 publications

(202 citation statements)

References 171 publications

(253 reference statements)

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“…Newly developed, sophisticated, superresolution microscopy techniques such as structured illumination microscopy, stimulated emission depletion microscopy, photoactivated localization microscopy, and stochastic optical reconstruction microscopy make it feasible to detect PPIs at the single-molecule level (Gutierrez et al, 2010;Komis et al, 2015;Sydor et al, 2015). In addition, spectromicroscopy techniques ) such as fluorescence cross-correlation spectroscopy (Macháň and Wohland, 2014) represent technological improvements that greatly improve the detection of the spatiotemporal resolution of interactions, particularly in (but not limited to) membrane environments.…”

Section: Discussionmentioning

confidence: 99%

Techniques for the analysis of protein-protein interactions in vivo

et al. 2016

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ORCID ID: 0000-0002-5820-4466 (C.G.).Identifying key players and their interactions is fundamental for understanding biochemical mechanisms at the molecular level. The ever-increasing number of alternative ways to detect protein-protein interactions (PPIs) speaks volumes about the creativity of scientists in hunting for the optimal technique. PPIs derived from single experiments or high-throughput screens enable the decoding of binary interactions, the building of large-scale interaction maps of single organisms, and the establishment of crossspecies networks. This review provides a historical view of the development of PPI technology over the past three decades, particularly focusing on in vivo PPI techniques that are inexpensive to perform and/or easy to implement in a state-of-the-art molecular biology laboratory. Special emphasis is given to their feasibility and application for plant biology as well as recent improvements or additions to these established techniques. The biology behind each method and its advantages and disadvantages are discussed in detail, as are the design, execution, and evaluation of PPI analysis. We also aim to raise awareness about the technological considerations and the inherent flaws of these methods, which may have an impact on the biological interpretation of PPIs. Ultimately, we hope this review serves as a useful reference when choosing the most suitable PPI technique.

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Section: Discussionmentioning

confidence: 99%

Techniques for the analysis of protein-protein interactions in vivo

et al. 2016

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“…Seminal work by Robert Hooke 2 and others in the 1600’s showed, for the first time, the microscopic world and laid the pathway to our modern deep understanding of the cell and the fundamental building blocks of life and disease. The modern microscope has evolved to include, amongst others, phase contrast, darkfield 3 , fluorescence 4 and super-resolution methodologies 5 . It is a complex instrument combining advances in optics, detection, light generation, computation and engineering.…”

Section: Introductionmentioning

confidence: 99%

EnLightenment: High resolution smartphone microscopy as an educational and public engagement platform

Wicks¹,

Cairns²,

Melnyk³

et al. 2017

Wellcome Open Res

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We developed a simple, cost-effective smartphone microscopy platform for use in educational and public engagement programs. We demonstrated its effectiveness, and potential for citizen science through a national imaging initiative, EnLightenment. The cost effectiveness of the instrument allowed for the program to deliver over 500 microscopes to more than 100 secondary schools throughout Scotland, targeting 1000’s of 12-14 year olds. Through careful, quantified, selection of a high power, low-cost objective lens, our smartphone microscope has an imaging resolution of microns, with a working distance of 3 mm. It is therefore capable of imaging single cells and sub-cellular features, and retains usability for young children. The microscopes were designed in kit form and provided an interdisciplinary educational tool. By providing full lesson plans and support material, we developed a framework to explore optical design, microscope performance, engineering challenges on construction and real-world applications in life sciences, biological imaging, marine biology, art, and technology. A national online imaging competition framed EnLightenment ; with over 500 high quality images submitted of diverse content, spanning multiple disciplines. With examples of cellular and sub-cellular features clearly identifiable in some submissions, we show how young public can use these instruments for research-level imaging applications, and the potential of the instrument for citizen science programs.

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“…Novel developments in optical microscopy technology and photophysics [1] made it possible to radically overcome the classical diffraction limit (ca. 200 nm laterally, 600 nm along the optical axis; also called the Abbe-limit [2], Equation 1) of conventional far-field microscopy.…”

Section: Introductionmentioning

confidence: 99%