Abstract:
:Remote focussing microscopy allows sharp, in-focus images to be acquired at high speed from outside of the focal plane of an objective lens without any agitation of the specimen. However, without careful optical alignment, the advantages of remote focussing microscopy could be compromised by the introduction of depth-dependent scaling artifacts. To achieve an ideal alignment in a point-scanning remote focussing microscope, the lateral (XY) scan mirror pair must be imaged onto the back focal plane of both the reference and imaging objectives, in a telecentric arrangement. However, for many commercial objective lenses, it can be difficult to accurately locate the position of the back focal plane. This paper investigates the impact of this limitation on the fidelity of three-dimensional data sets of living cardiac tissue, specifically the introduction of distortions. These distortions limit the accuracy of sarcomere measurements taken directly from raw volumetric data. The origin of the distortion is first identified through simulation of a remote focussing microscope. Using a novel three-dimensional calibration specimen it was then possible to quantify experimentally the size of the distortion as a function of objective misalignment. Finally, by first approximating and then compensating the distortion in imaging data from whole heart rodent studies, the variance of sarcomere length (SL) measurements was reduced by almost 50%.
journal_name
Front Physioljournal_title
Frontiers in physiologyauthors
Corbett AD,Burton RA,Bub G,Salter PS,Tuohy S,Booth MJ,Wilson Tdoi
10.3389/fphys.2014.00384subject
Has Abstractpub_date
2014-10-08 00:00:00pages
384issn
1664-042Xjournal_volume
5pub_type
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journal_title:Frontiers in physiology
pub_type: 杂志文章,评审
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journal_title:Frontiers in physiology
pub_type: 杂志文章,评审
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abstract::[This corrects the article DOI: 10.3389/fphys.2020.00697.]. ...
journal_title:Frontiers in physiology
pub_type: 已发布勘误
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pub_type: 杂志文章,评审
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pub_type: 杂志文章
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pub_type: 杂志文章,评审
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pub_type: 杂志文章,评审
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pub_type: 杂志文章
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pub_type: 杂志文章
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abstract::Circadian rhythms are biological oscillations with a period of ~24 h. These rhythms are orchestrated by a circadian timekeeper in the suprachiasmatic nucleus of the hypothalamus, the circadian "master clock," which exactly adjusts clock outputs to solar time via photic synchronization. At the molecular level, circadia...
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更新日期:2020-02-21 00:00:00
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doi:10.3389/fphys.2013.00245
更新日期:2013-09-12 00:00:00
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journal_title:Frontiers in physiology
pub_type: 杂志文章
doi:10.3389/fphys.2017.00142
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journal_title:Frontiers in physiology
pub_type: 杂志文章
doi:10.3389/fphys.2016.00166
更新日期:2016-05-09 00:00:00
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pub_type: 杂志文章,评审
doi:10.3389/fphys.2018.01158
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journal_title:Frontiers in physiology
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pub_type: 杂志文章
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pub_type: 杂志文章,评审
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pub_type: 杂志文章
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