Optical sectioning microscopy

Conchello, José-Angel; Lichtman, Jeff W.

doi:10.1038/nmeth815

Cited by 661 publications

(447 citation statements)

References 43 publications

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“…The axon diameter cannot be measured directly by optical microscopy because it is often Ͻ1 m and smaller than the point spread function (Conchello and Lichtman, 2005). Therefore, by assuming that the total fluorescence intensity ( F) in a given axonal segment is proportional to the axonal volume [i.e., F ϰ volume ϭ (d/2) 2 ϫ unit length], we estimated the relative axon diameter as the square root of the Alexa Fluor 488 fluorescence intensity, i.e., d ϰ F 1/2 .…”

Section: Computational Prediction Of Somatic Ap Modulationmentioning

confidence: 99%

Effects of Axonal Topology on the Somatic Modulation of Synaptic Outputs

Sasaki

Matsuki²,

Ikegaya³

2012

J. Neurosci.

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Depolarization of the neuronal soma augments synaptic output onto postsynaptic neurons via long-range, axonal cable properties. Here, we report that the range of this somatic influence is spatially restricted by not only axonal path length but also a branching-dependent decrease in axon diameter. Cell-attached recordings of action potentials (APs) from multiple axon branches of a rat hippocampal CA3 pyramidal cell revealed that an AP was broadened following a 20 mV depolarization of the soma and reverted to a normal width during propagation down the axon. The narrowing of the AP depended on the distance traveled by the AP and on the number of axon branch points through which the AP passed. These findings were confirmed by optical imaging of AP-induced calcium elevations in presynaptic boutons, suggesting that the somatic membrane potential modifies synaptic outputs near the soma but not long-projection outputs. Consistent with this prediction, whole-cell recordings from synaptically connected neurons revealed that depolarization of presynaptic CA3 pyramidal cells facilitated synaptic transmission to nearby CA3 pyramidal cells, but not to distant pyramidal cells in CA3 or CA1. Therefore, axonal geometry enables the differential modulation of synaptic output depending on target location.

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Section: Computational Prediction Of Somatic Ap Modulationmentioning

confidence: 99%

Effects of Axonal Topology on the Somatic Modulation of Synaptic Outputs

Sasaki

Matsuki²,

Ikegaya³

2012

J. Neurosci.

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“…In light optics, there exists a large body of work concerning the deconvolution of the blurring caused by imaging systems in three dimensions for both confocal and widefield microscopes (McNally et al 1999;Conchello & Lichtman 2005). At present, confocal imaging is the most common method owing to its optical sectioning capabilities.…”

Section: (C) Deconvolution Of Experimental Imagesmentioning

confidence: 99%

Three-dimensional imaging by optical sectioning in the aberration-corrected scanning transmission electron microscope

Behan

Cosgriff

Kirkland

et al. 2009

Phil. Trans. R. Soc. A.

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The depth resolution for optical sectioning in the scanning transmission electron microscope is measured using the results of optical sectioning experiments of laterally extended objects. We show that the depth resolution depends on the numerical aperture of the objective lens as expected. We also find, however, that the depth resolution depends on the lateral extent of the object that is being imaged owing to a missing cone of information in the transfer function. We find that deconvolution methods generally have limited usefulness in this case, but that three-dimensional information can still be obtained with the aid of prior information for specific samples such as those consisting of supported nanoparticles. We go on to review how a confocal geometry may improve the depth resolution for extended objects. Finally, we present a review of recent work exploring the effect of dynamical diffraction in zone-axis-aligned crystals on the optical sectioning process.

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“…Today, a variety of methods are available for optical sectioning microscopy [1]. They can be grouped in three large categories: a) techniques based purely on optical means to prevent outof-focus light from being detected, b) techniques which rely purely on computational means to subsequently remove out-of-focus light from three dimensional widefield data sets, and c) techniques using optical means to encode additional information in the image which can then be used to greatly facilitate computation of an optical section from the raw data.…”

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confidence: 99%

Structure brings clarity: Structured illumination microscopy in cell biology

Langhorst

Schaffer

Goetze

2009

Biotechnology Journal

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International audienceBiological samples are three dimensional and therefore optical sectioning is mandatory for microscopic images to precisely show the localization or function of structures within biological samples. Today, researchers can choose from a variety of methods to obtain optical sections. This article focuses on structured illumination microscopy, which is a group of techniques utilizing a combination of optics and mathematics to obtain optical sections: A structure is imaged onto the sample by optical means and the additional information thereby encoded in the image is used to calculate an optical section from several acquired images. Different methods of structured illumination microscopy (mainly grid projection and aperture correlation) are discussed from a practical point of view, concentrating on advantages, limitations and future prospects of these techniques and their use in cell biology. Structured illumination can also be used to obtain superresolution information if structures of higher frequency are projected onto the sample. This promising approach to superresolution microscopy is also briefly discussed from a user's perspective

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Optical sectioning microscopy

Cited by 661 publications

References 43 publications

Effects of Axonal Topology on the Somatic Modulation of Synaptic Outputs

Effects of Axonal Topology on the Somatic Modulation of Synaptic Outputs

Three-dimensional imaging by optical sectioning in the aberration-corrected scanning transmission electron microscope

Structure brings clarity: Structured illumination microscopy in cell biology

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