Superresolution microscopy for microbiology
Carla Coltharp,Jie Xiao +1 more
TL;DR: The goal of this review is to provide the necessary technical background for interested microbiologists to choose the appropriate superresolution method for their biological systems, and to introduce the practical considerations required for designing and analysing superresolution imaging experiments.
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Abstract: This review provides a practical introduction to superresolution microscopy from the perspective of microbiological research. Because of the small sizes of bacterial cells, superresolution methods are particularly powerful and suitable for revealing details of cellular structures that are not resolvable under conventional fluorescence light microscopy. Here we describe the methodological concepts behind three major categories of superresolution light microscopy: photoactivated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM), structured illumination microscopy (SIM) and stimulated emission-depletion (STED) microscopy. We then present recent applications of each of these techniques to microbial systems, which have revealed novel conformations of cellular structures and described new properties of in vivo protein function and interactions. Finally, we discuss the unique issues related to implementing each of these superresolution techniques with bacterial specimens and suggest avenues for future development. The goal of this review is to provide the necessary technical background for interested microbiologists to choose the appropriate superresolution method for their biological systems, and to introduce the practical considerations required for designing and analysing superresolution imaging experiments.
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Citations
Three-Dimensional Localization of Single Molecules for Super-Resolution Imaging and Single-Particle Tracking
TL;DR: In this article, a variety of methods for obtaining both 3D super-resolution images and 3D tracking infers structures or motions extending in the axial direction can easily be missed or confused.
Fluorescent proteins for live-cell imaging with super-resolution
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Analysing Microbial Community Composition through Amplicon Sequencing: From Sampling to Hypothesis Testing.
TL;DR: This review gives a historical perspective on the use of sequencing data in microbial ecology and restate the current need for this method; but also highlights the major caveats with standard practices for handling these data, from sample collection and library preparation to statistical analysis.
Exploring bacterial cell biology with single-molecule tracking and super-resolution imaging
Andreas Gahlmann,W. E. Moerner +1 more
TL;DR: This Review considers what single-molecule fluorescence tracking and super-resolution imaging have taught us about the bacterial cytoskeleton, nucleoid organization and the dynamic processes of transcription and translation, and highlights the methodological improvements that are needed to address a number of experimental challenges.
285
Super-Resolution Microscopy: From Single Molecules to Supramolecular Assemblies
TL;DR: This review surveys the application of SRM in elucidating the structure of macromolecules in the native cellular environment and discusses both the novel information that can be generated through SRM as well as the experimental considerations to examine while conducting such studies.
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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.
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