Eight years of single-molecule localization microscopy

Klein, Teresa; Proppert, Sven; Sauer, Markus

doi:10.1007/s00418-014-1184-3

Cited by 140 publications

(113 citation statements)

References 124 publications

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“…iPALM identifies a molecule's z position using the interference of the molecule's emitted light along two optical paths and provides the best z resolution currently available to fluorescence-based superresolution imaging (56). Under our experimental condition using the mEos2 fusion protein, we achieved spatial resolutions of ∼30 nm, ∼24 nm, and ∼16 nm in the x, y, and z dimensions, respectively (SI Appendix, Fig.…”

Section: Resultsmentioning

confidence: 96%

Defining the rate-limiting processes of bacterial cytokinesis

Coltharp

Buss

Plumer

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

160

216

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Bacterial cytokinesis is accomplished by the essential ‘divisome’ machinery. The most widely conserved divisome component, FtsZ, is a tubulin homolog that polymerizes into the ‘FtsZ-ring’ (‘Z-ring’). Previous in vitro studies suggest that Z-ring contraction serves as a major constrictive force generator to limit the progression of cytokinesis. Here, we applied quantitative superresolution imaging to examine whether and how Z-ring contraction limits the rate of septum closure during cytokinesis in Escherichia coli cells. Surprisingly, septum closure rate was robust to substantial changes in all Z-ring properties proposed to be coupled to force generation: FtsZ’s GTPase activity, Z-ring density, and the timing of Z-ring assembly and disassembly. Instead, the rate was limited by the activity of an essential cell wall synthesis enzyme and further modulated by a physical divisome–chromosome coupling. These results challenge a Z-ring–centric view of bacterial cytokinesis and identify cell wall synthesis and chromosome segregation as limiting processes of cytokinesis.

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Section: Resultsmentioning

confidence: 96%

Defining the rate-limiting processes of bacterial cytokinesis

Coltharp

Buss

Plumer

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

160

216

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“…With this technique, a lateral resolution of 20 nm is typical. 35,37 We were able to identify distinct microtubules in the ventral nerve cord of C. elegans in electron micrographs as well as in dSTORM images (Fig. 8).…”

Section: Correlation Using Intrinsic Landmarks Is Easilymentioning

confidence: 89%

Filling the gap: adding super-resolution to array tomography for correlated ultrastructural and molecular identification of electrical synapses at theC. elegansconnectome

et al. 2016

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Abstract. Correlating molecular labeling at the ultrastructural level with high confidence remains challenging. Array tomography (AT) allows for a combination of fluorescence and electron microscopy (EM) to visualize subcellular protein localization on serial EM sections. Here, we describe an application for AT that combines near-native tissue preservation via high-pressure freezing and freeze substitution with super-resolution light microscopy and high-resolution scanning electron microscopy (SEM) analysis on the same section. We established protocols that combine SEM with structured illumination microscopy (SIM) and direct stochastic optical reconstruction microscopy (dSTORM). We devised a method for easy, precise, and unbiased correlation of EM images and super-resolution imaging data using endogenous cellular landmarks and freely available image processing software. We demonstrate that these methods allow us to identify and label gap junctions in Caenorhabditis elegans with precision and confidence, and imaging of even smaller structures is feasible. With the emergence of connectomics, these methods will allow us to fill in the gap-acquiring the correlated ultrastructural and molecular identity of electrical synapses.

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“…Volume-rendering imaging techniques such as confocal microscopy provide fine details of structures in multi-dimensions superior to those obtained with conventional widefield microscopy, yet still plagued by diffraction-limited resolution (Conchello and Lichtman 2006). Super-resolution microscopy techniques overcome the light microscopy diffraction limit, but still do not afford the superior resolution attained by electron microscopy (Klein et al 2014). Thus, to perform a highresolution three-dimensional reconstruction of the intercalated disc in the myocardium, Vanslembrouck et al (2018) turned to volume producing scanning electron microscopy (SEM) techniques.…”

Section: Volume Electron Microscopy Methods To Study Cardiomyocyte Inmentioning

confidence: 99%

In focus in HCB

Taatjes

Roth

2018

Histochem Cell Biol

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Blood-brain barrier computational pathologyIn the first of the review articles commemorating the 60th anniversary of Histochemistry and Cell Biology, Shityakov and Förster (2018) present a timely update on in silico modeling of the blood-brain barrier (BBB) in a variety of pathological circumstances. After initially describing some computational models employed to investigate BBB pathology in general, they then proceed to describe in individual sections specific computer models effective for the analysis of BBB issues, progressing from the molecular, to the cellular, tissue, and organ levels. Moreover, they provide a very informative table presenting a summary of the computational models currently in use at all levels of resolution relating to BBB pathology. The described models are based on mathematical principals and algorithms, in conjunction with multi-dimensional experimental data sets. Though much molecular information is currently available concerning proteins expressed at the BBB, much remains to be learned concerning their alterations in pathological conditions, such as traumatic brain injury. The authors finish with a thoughtful synopsis on their perspectives for future areas and methods of investigation in this critical area of CNS pathology.

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Eight years of single-molecule localization microscopy

Cited by 140 publications

References 124 publications

Defining the rate-limiting processes of bacterial cytokinesis

Defining the rate-limiting processes of bacterial cytokinesis

Filling the gap: adding super-resolution to array tomography for correlated ultrastructural and molecular identification of electrical synapses at theC. elegansconnectome

In focus in HCB

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