Incorporation of an experimentally determined MTF for spatial frequency filtering and deconvolution during optical projection tomography reconstruction
Lingling Chen,James McGinty,Harriet B. Taylor,Laurence Bugeon,Jonathan R. Lamb,Margaret J. Dallman,Paul M. W. French +6 more
TLDR
Two techniques to improve the quality of reconstructed optical projection tomography (OPT) images using the modulation transfer function (MTF) as a function of defocus experimentally determined from tilted knife-edge measurements are demonstrated.Abstract:
We demonstrate two techniques to improve the quality of reconstructed optical projection tomography (OPT) images using the modulation transfer function (MTF) as a function of defocus experimentally determined from tilted knife-edge measurements. The first employs a 2-D binary filter based on the MTF frequency cut-off as an additional filter during back-projection reconstruction that restricts the high frequency information to the region around the focal plane and progressively decreases the spatial frequency bandwidth with defocus. This helps to suppress “streak” artifacts in OPT data acquired at reduced angular sampling, thereby facilitating faster OPT acquisitions. This method is shown to reduce the average background by approximately 72% for an NA of 0.09 and by approximately 38% for an NA of 0.07 compared to standard filtered back-projection. As a biological illustration, a Fli:GFP transgenic zebrafish embryo (3 days post-fertilisation) was imaged to demonstrate the improved imaging speed (a quarter of the acquisition time). The second method uses the MTF to produce an appropriate deconvolution filter that can be used to correct for the spatial frequency modulation applied by the imaging system.read more
Citations
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Journal ArticleDOI
Helical optical projection tomography.
TL;DR: 3D-reconstructed images of the colon of DSS-treated mice, a model for Inflammatory Bowel Disease, allowed the identification of the structural alterations and the geometry of the hOPT device facilitates the addition of a Selective Plane Illumination Microscopy (SPIM) arm, providing the possibility of delivering high resolution images of selected areas together with complete volumetric information.
Journal ArticleDOI
Visualising apoptosis in live zebrafish using fluorescence lifetime imaging with optical projection tomography to map FRET biosensor activity in space and time
Natalie Andrews,Marie-Christine Ramel,Marie-Christine Ramel,Sunil Kumar,Yuriy Alexandrov,Douglas J. Kelly,Sean C. Warren,Louise Kerry,Nicola Lockwood,Antonina Frolov,Paul Frankel,Laurence Bugeon,James McGinty,Margaret J. Dallman,Paul M. W. French +14 more
TL;DR: The first application of FLIM OPT to read out biological function in live transgenic zebrafish larvae using a genetically expressed FRET biosensor is reported, enabling apoptosis to be monitored over time via changes in Caspase 3 activation following gamma irradiation at 24 hours post fertilisation.
Journal ArticleDOI
Simultaneous angular multiplexing optical projection tomography at shifted focal planes
TL;DR: It is demonstrated that imaging at two orthogonal angular projections simultaneously, focused on shifted planes in the sample, improves the average spatial resolution by ~20% and the light collection efficiency by a factor of ~4, thereby enabling increased acquisition speed and reduced light dose.
Journal ArticleDOI
Applicability, usability, and limitations of murine embryonic imaging with optical coherence tomography and optical projection tomography.
Manmohan Singh,Raksha Raghunathan,Victor G. Piazza,Anjul M. Davis-Loiacono,Alex Cable,Tegy J. Vedakkan,Trevor Janecek,Michael V. Frazier,Achuth Nair,Chen Wu,Irina V. Larina,Mary E. Dickinson,Kirill V. Larin +12 more
TL;DR: It is demonstrated that while OCT was capable of rapid high-resolution live 3D imaging, its limited penetration depth prevented visualization of deeper structures, particularly in later stage embryos, and OPT could not be used in vivo because the embryos must be fixed and cleared.
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Assessment of optical CT as a future QA tool for synchrotron x-ray microbeam therapy
TL;DR: The extent to which new developments in 3D dosimetry using optical computed tomography (CT) can visualise MRT dose distributions is investigated, and what further developments are necessary are assessed before fully quantitative 3D measurements can be achieved.
References
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Two-Photon Laser Scanning Fluorescence Microscopy
TL;DR: The fluorescence emission increased quadratically with the excitation intensity so that fluorescence and photo-bleaching were confined to the vicinity of the focal plane as expected for cooperative two-photon excitation.
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Optical sectioning deep inside live embryos by selective plane illumination microscopy
TL;DR: In this article, a selective plane illumination microscopy (SPIM) was developed to generate multidimensional images of samples up to a few millimeters in size, which can be applied to visualize the embryogenesis of the relatively opaque Drosophila melanogaster in vivo.
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Optical Projection Tomography as a Tool for 3D Microscopy and Gene Expression Studies
James Sharpe,Ulf Ahlgren,Paul Perry,Bill Hill,Allyson Ross,Jacob Hecksher-Sørensen,Richard Baldock,Duncan Davidson +7 more
TL;DR: This work has developed a microscopy technique that uses optical projection tomography (OPT) to produce high-resolution 3D images of both fluorescent and nonfluorescent biological specimens with a thickness of up to 15 millimeters.
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Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain.
Hans-Ulrich Dodt,Ulrich Leischner,Anja Schierloh,Nina Jährling,Nina Jährling,Christoph P. Mauch,Katrin Deininger,Jan M. Deussing,Matthias Eder,Walter Zieglgänsberger,Klaus Becker,Klaus Becker +11 more
TL;DR: This new technique allows optical sectioning of fixed mouse brains with cellular resolution and can be used to detect single GFP-labeled neurons in excised mouse hippocampi and is ideally suited for high-throughput phenotype screening of transgenic mice and thus will benefit the investigation of disease models.