Aptamers as potential tools for super-resolution microscopy

Opazo, Felipe; Levy, Matthew; Byrom, Michelle; Schäfer, Christina; Geisler, Claudia; Groemer, Teja W.; Ellington, Andrew D.; Rizzoli, Silvio O.

doi:10.1038/nmeth.2179

Cited by 163 publications

(164 citation statements)

References 5 publications

(4 reference statements)

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“…Blom et al 2011;Kellner et al 2007;Kittel et al 2006;Lau et al 2012;Muller et al 2012;Opazo et al 2012;Persson et al 2011;Schmidt et al 2008Schmidt et al , 2009Sieber et al 2007;Wagner et al 2012), making a STED microscope a uniquely helpful tool nowadays in cell-biological laboratories (Clausen et al 2013). Furthermore, STED has important applications outside biology, ranging from nanoscale imaging of assemblies of colloidal particles and polymeric structures (Friedemann et al 2011;Harke et al 2008b;Ullal et al 2009Ullal et al , 2011 to solid-state physics .…”

Section: Sted Nanoscopymentioning

confidence: 99%

“…In general, coming along with the increased sensitivity of the nanoscopy approach, greater care has to be taken when labelling cellular samples, especially with respect to unspecific background staining . Artifacts due to, for example, improper fixation in immunolabeling or unspecific binding may not be observed in confocal but may be visible in nanoscopy images, due to the improved spatial resolution in the latter (Opazo et al 2012;Tanaka et al 2010). Also, due to the increased spatial resolution, the term "co-localization" may become invalid, since two objects (especially when labelled via a primary and secondary antibody as in most immunolabeling approaches) cannot occupy the same spot.…”

Section: Conclusion: Coordinate-targeted Versus -Stochasticmentioning

confidence: 99%

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Lens-based fluorescence nanoscopy

Eggeling

Willig

Sahl

et al. 2015

Quart. Rev. Biophys.

137

139

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The majority of studies of the living cell rely on capturing images using fluorescence microscopy. Unfortunately, for centuries, diffraction of light was limiting the spatial resolution in the optical microscope: structural and molecular details much finer than about half the wavelength of visible light (~200 nm) could not be visualized, imposing significant limitations on this otherwise so promising method. The surpassing of this resolution limit in far-field microscopy is currently one of the most momentous developments for studying the living cell, as the move from microscopy to super-resolution microscopy or 'nanoscopy' offers opportunities to study problems in biophysical and biomedical research at a new level of detail. This review describes the principles and modalities of present fluorescence nanoscopes, as well as their potential for biophysical and cellular experiments. All the existing nanoscopy variants separate neighboring features by transiently preparing their fluorescent molecules in states of different emission characteristics in order to make the features discernible. Usually these are fluorescent 'on' and 'off' states causing the adjacent molecules to emit sequentially in time. Each of the variants can in principle reach molecular spatial resolution and has its own advantages and disadvantages. Some require specific transitions and states that can be found only in certain fluorophore subfamilies, such as photoswitchable fluorophores, while other variants can be realized with standard fluorescent labels. Similar to conventional far-field microscopy, nanoscopy can be utilized for dynamical, multi-color and three-dimensional imaging of fixed and live cells, tissues or organisms. Lens-based fluorescence nanoscopy is poised for a high impact on future developments in the life sciences, with the potential to help solve long-standing quests in different areas of scientific research.

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Section: Sted Nanoscopymentioning

confidence: 99%

Section: Conclusion: Coordinate-targeted Versus -Stochasticmentioning

confidence: 99%

Lens-based fluorescence nanoscopy

Eggeling

Willig

Sahl

et al. 2015

Quart. Rev. Biophys.

137

139

View full text Add to dashboard Cite

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“…While polyclonal antibodies can lead to overlabeling, the large size of the antibody can also lead to underlabeling due to steric hindrance. In practice, determining the labeling efficiency of an antibody is quite challenging and smaller probes such as aptamers and nanobodies have been demonstrated to lead to denser labeling of certain targets and higher spatial resolution in super-resolution images [38,39]. In principle, fluorescent proteins avoid this complication, giving rise to a one-to-one labeling ratio; however, the expression strategy used can cause additional problems.…”

Section: Missed Molecules and Under-countingmentioning

confidence: 99%

Quantitative super-resolution microscopy: pitfalls and strategies for image analysis

Durisic

Laparra-Cuervo

Lakadamyali

2014

Current Opinion in Chemical Biology

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Super-resolution microscopy is an enabling technology that allows biologists to visualize cellular structures at nanometer length scales using far-field optics. To break the diffraction barrier, it is necessary to leverage the distinct molecular states of fluorescent probes. At the same time, the existence of these different molecular states and the photophysical properties of the fluorescent probes can complicate data quantification and interpretation. Here, we review the pitfalls in super-resolution data analysis that must be avoided for proper interpretation of images.

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“…57 Single-chain antibody fragments (nanobodies) against tubulin was engineered to achieve super-resolution imaging of microtubules with a decreased apparent diameter. 58 In parallel, aptamers (short single stranded oligonucleotides ($ 15 kDa, $ 4 nm), as small as nanobodies, can also bene¯t the STED imaging when compared with antibody staining, 59 as seen in Fig. 3.…”

Section: Emerging Small Molecular Labeling Methodsmentioning

confidence: 99%

Super-resolution imaging in glycoscience: New developments and challenges

Chen

Tong

Wang

2016

J. Innov. Opt. Health Sci.

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Carbohydrates on cell surfaces play a crucial role in a wide variety of biological processes, including cell adhesion, recognition and signaling, viral and bacterial infection, in°ammation and metastasis. However, owing to the large diversity and complexity of carbohydrate structure and nongenetically synthesis, glycoscience is the least understood¯eld compared with genomics and proteomics. Although the structures and functions of carbohydrates have been investigated by various conventional analysis methods, the distribution and role of carbohydrates in cell membranes remain elusive. This review focuses on the developments and challenges of super-resolution imaging in glycoscience through introduction of imaging principle and the available°uorescent probes for super-resolution imaging, the labeling strategies of carbohydrates, and the recent applications of super-resolution imaging in glycoscience, which will promote the super-resolution imaging technology as a promising tool to provide new insights into the study of glycoscience.

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Aptamers as potential tools for super-resolution microscopy

Cited by 163 publications

References 5 publications

Lens-based fluorescence nanoscopy

Lens-based fluorescence nanoscopy

Quantitative super-resolution microscopy: pitfalls and strategies for image analysis

Super-resolution imaging in glycoscience: New developments and challenges

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