Author
Philippe Roudot
Other affiliations: University of Texas at Dallas, French Institute for Research in Computer Science and Automation
Bio: Philippe Roudot is an academic researcher from University of Texas Southwestern Medical Center. The author has contributed to research in topics: Light sheet fluorescence microscopy & Optical sectioning. The author has an hindex of 12, co-authored 32 publications receiving 1209 citations. Previous affiliations of Philippe Roudot include University of Texas at Dallas & French Institute for Research in Computer Science and Automation.
Papers
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Max Planck Society1, Centre national de la recherche scientifique2, University of Navarra3, Masaryk University4, Drexel University5, Zhejiang University6, Royal Institute of Technology7, Curie Institute8, French Institute for Research in Computer Science and Automation9, Pasteur Institute10, Leiden University11, Chung Yuan Christian University12
TL;DR: Although no single method performed best across all scenarios, the results revealed clear differences between the various approaches, leading to notable practical conclusions for users and developers.
Abstract: Particle tracking is of key importance for quantitative analysis of intracellular dynamic processes from time-lapse microscopy image data. Because manually detecting and following large numbers of individual particles is not feasible, automated computational methods have been developed for these tasks by many groups. Aiming to perform an objective comparison of methods, we gathered the community and organized an open competition in which participating teams applied their own methods independently to a commonly defined data set including diverse scenarios. Performance was assessed using commonly defined measures. Although no single method performed best across all scenarios, the results revealed clear differences between the various approaches, leading to notable practical conclusions for users and developers.
819 citations
TL;DR: A light sheet fluorescence microscope that achieves 390 nm isotropic resolution and high optical sectioning strength over large field of views, without the need for structured illumination or deconvolution-based postprocessing is presented.
185 citations
TL;DR: Cleared-tissue axially swept light-sheet microscopy (ctASLM) enables high-speed, refraction index-independent imaging of live, cleared and expanded samples with isotropic, submicron resolution.
Abstract: We present cleared-tissue axially swept light-sheet microscopy (ctASLM), which enables isotropic, subcellular resolution imaging with high optical sectioning capability and a large field of view over a broad range of immersion media. ctASLM can image live, expanded, and both aqueous and non-aqueous chemically cleared tissue preparations. Depending on the optical configuration, ctASLM provides up to 260 nm of axial resolution, a three to tenfold improvement over confocal and other reported cleared-tissue light-sheet microscopes. We imaged millimeter-scale cleared tissues with subcellular three-dimensional resolution, which enabled automated detection of multicellular tissue architectures, individual cells, synaptic spines and rare cell–cell interactions. Cleared-tissue axially swept light-sheet microscopy (ctASLM) enables high-speed, refraction index-independent imaging of live, cleared and expanded samples with isotropic, submicron resolution.
124 citations
TL;DR: It is shown that Augmin-dependent, noncentrosomal nucleation generates the vast majority of microtubules in metaphase spindles, which results in a strong directional bias of micro Tubule growth toward individual kinetochores.
Abstract: Dividing cells reorganize their microtubule cytoskeleton into a bipolar spindle, which moves one set of sister chromatids to each nascent daughter cell. Early spindle assembly models postulated that spindle pole-derived microtubules search the cytoplasmic space until they randomly encounter a kinetochore to form a stable attachment. More recent work uncovered several additional, centrosome-independent microtubule generation pathways, but the contributions of each pathway to spindle assembly have remained unclear. Here, we combined live microscopy and mathematical modeling to show that most microtubules nucleate at noncentrosomal regions in dividing human cells. Using a live-cell probe that selectively labels aged microtubule lattices, we demonstrate that the distribution of growing microtubule plus ends can be almost entirely explained by Augmin-dependent amplification of long-lived microtubule lattices. By ultrafast 3D lattice light-sheet microscopy, we observed that this mechanism results in a strong directional bias of microtubule growth toward individual kinetochores. Our systematic quantification of spindle dynamics reveals highly coordinated microtubule growth during kinetochore fiber assembly.
72 citations
TL;DR: The field synthesis method for generating any scanned light sheet is based on a new mathematical theorem in Fourier analysis and has important practical implications for simpler, multicolor lattice light-sheet microscopy.
Abstract: We introduce field synthesis, a theorem and method that can be used to synthesize any scanned or dithered light sheet, including those used in lattice light-sheet microscopy (LLSM), from an incoherent superposition of one-dimensional intensity distributions. Compared to LLSM, this user-friendly and modular approach offers a simplified optical design, higher light throughput and simultaneous multicolor illumination. Further, field synthesis achieves lower rates of photobleaching than light sheets generated by lateral beam scanning.
61 citations
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TL;DR: TrackMate is an extensible platform where developers can easily write their own detection, particle linking, visualization or analysis algorithms within the TrackMate environment and is validated for quantitative lifetime analysis of clathrin-mediated endocytosis in plant cells.
2,356 citations
01 Jan 2016
TL;DR: In this paper, the authors present the principles of optics electromagnetic theory of propagation interference and diffraction of light, which can be used to find a good book with a cup of coffee in the afternoon, instead of facing with some infectious bugs inside their computer.
Abstract: Thank you for reading principles of optics electromagnetic theory of propagation interference and diffraction of light. As you may know, people have search hundreds times for their favorite novels like this principles of optics electromagnetic theory of propagation interference and diffraction of light, but end up in harmful downloads. Rather than enjoying a good book with a cup of coffee in the afternoon, instead they are facing with some infectious bugs inside their computer.
2,213 citations
Journal Article•
1,261 citations
TL;DR: This work elucidate the key developments and define a simple set of underlying principles governing LSFM, which aim to clarify the decisions to be made for those who wish to develop and use bespoke light-sheet systems and to assist in identifying the best approaches to apply this powerful technique to myriad biological questions.
Abstract: This Review introduces the fundamental considerations for building a light sheet microscope, describes the pros and cons associated with available implementations, and offers practical advice for users.
484 citations