Journal ArticleDOI
Fluorescence nanoscopy in cell biology
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TLDR
In this Review,luorescence nanoscopy uniquely combines minimally invasive optical access to the internal nanoscale structure and dynamics of cells and tissues with molecular detection specificity and the labelling of individual molecules to enable their visualization has emerged as a central challenge.Abstract:
Fluorescence nanoscopy uniquely combines minimally invasive optical access to the internal nanoscale structure and dynamics of cells and tissues with molecular detection specificity. While the basic physical principles of 'super-resolution' imaging were discovered in the 1990s, with initial experimental demonstrations following in 2000, the broad application of super-resolution imaging to address cell-biological questions has only more recently emerged. Nanoscopy approaches have begun to facilitate discoveries in cell biology and to add new knowledge. One current direction for method improvement is the ambition to quantitatively account for each molecule under investigation and assess true molecular colocalization patterns via multi-colour analyses. In pursuing this goal, the labelling of individual molecules to enable their visualization has emerged as a central challenge. Extending nanoscale imaging into (sliced) tissue and whole-animal contexts is a further goal. In this Review we describe the successes to date and discuss current obstacles and possibilities for further development.read more
Citations
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Journal ArticleDOI
Super-resolution microscopy demystified
Lothar Schermelleh,Alexia Ferrand,Thomas R Huser,Christian Eggeling,Markus Sauer,Oliver Biehlmaier,Drummen Gpc. +6 more
TL;DR: An overview of current super-resolution microscopy techniques is given and guidance on how best to use them to foster biological discovery is provided.
Journal ArticleDOI
Visualizing and discovering cellular structures with super-resolution microscopy
TL;DR: An overview of super-resolution methods, their state-of-the-art capabilities, and their constantly expanding applications to biology are provided, with a focus on the latter.
Journal ArticleDOI
Reactive Oxygen Species and Mitochondrial Dynamics: The Yin and Yang of Mitochondrial Dysfunction and Cancer Progression
TL;DR: The latest findings on the intricate relationship between mitochondrial dynamics and ROS production are reviewed, focusing mainly on its role in malignant disease.
Journal ArticleDOI
Small-Molecule Fluorescent Probes for Live-Cell Super-Resolution Microscopy.
Lu Wang,Michelle S. Frei,Michelle S. Frei,Aleksandar Salim,Aleksandar Salim,Kai Johnsson,Kai Johnsson +6 more
TL;DR: The role of small-molecule fluorescent probes for live-cell super-resolution microscopy and the challenges that need to be overcome for their generation are discussed and recent trends in the development of labeling strategies are reviewed.
Journal ArticleDOI
FRET as a biomolecular research tool - understanding its potential while avoiding pitfalls.
TL;DR: Forster resonance energy transfer (FRET) applications are not consistently applied, nor are results uniformly presented, which makes implementing and reproducing FRET experiments challenging as discussed by the authors. But FRET can be a powerful research tool.
References
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Journal ArticleDOI
Imaging intracellular fluorescent proteins at nanometer resolution.
Eric Betzig,George H. Patterson,Rachid Sougrat,O. Wolf Lindwasser,Scott G. Olenych,Juan S. Bonifacino,Michael W. Davidson,Jennifer Lippincott-Schwartz,Harald F. Hess +8 more
TL;DR: This work introduced a method for optically imaging intracellular proteins at nanometer spatial resolution and used this method to image specific target proteins in thin sections of lysosomes and mitochondria and in fixed whole cells to image retroviral protein Gag at the plasma membrane.
Journal ArticleDOI
Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM).
TL;DR: A high-resolution fluorescence microscopy method based on high-accuracy localization of photoswitchable fluorophores that can, in principle, reach molecular-scale resolution is developed.
Journal ArticleDOI
Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy
Stefan W. Hell,Jan Wichmann +1 more
TL;DR: A new type of scanning fluorescence microscope capable of resolving 35 nm in the far field is proposed, overcome the diffraction resolution limit by employing stimulated emission to inhibit the fluorescence process in the outer regions of the excitation point-spread function.
Journal Article
Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy
Stefan W. Hell,Jan Wichmann +1 more
TL;DR: In this paper, the authors proposed a new type of scanning fluorescence microscope capable of resolving 35 nm in the far field by employing stimulated emission to inhibit the fluorescence process in the outer regions of the excitation point spread function.
Journal ArticleDOI
Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy
TL;DR: A new method for fluorescence imaging has been developed that can obtain spatial distributions of large numbers of fluorescent molecules on length scales shorter than the classical diffraction limit, and suggests a means to address a significant number of biological questions that had previously been limited by microscope resolution.
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