Multiview tiling light sheet microscopy for 3D high-resolution live imaging. 2021

Mostafa Aakhte, and Hans-Arno J Müller
Developmental Genetics Group, Institute of Biology, University of Kassel, Heinrich-Plett Strasse 40, 34132 Kassel, Germany.

Light-sheet or selective plane illumination microscopy (SPIM) is ideally suited for in toto imaging of living specimens at high temporal-spatial resolution. In SPIM, the light scattering that occurs during imaging of opaque specimens brings about limitations in terms of resolution and the imaging field of view. To ameliorate this shortcoming, the illumination beam can be engineered into a highly confined light sheet over a large field of view and multi-view imaging can be performed by applying multiple lenses combined with mechanical rotation of the sample. Here, we present a Multiview tiling SPIM (MT-SPIM) that combines the Multi-view SPIM (M-SPIM) with a confined, multi-tiled light sheet. The MT-SPIM provides high-resolution, robust and rotation-free imaging of living specimens. We applied the MT-SPIM to image nuclei and Myosin II from the cellular to subcellular spatial scale in early Drosophila embryogenesis. We show that the MT-SPIM improves the axial-resolution relative to the conventional M-SPIM by a factor of two. We further demonstrate that this axial resolution enhancement improves the automated segmentation of Myosin II distribution and of nuclear volumes and shapes.

UI MeSH Term Description Entries
D008856 Microscopy, Fluorescence Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye. Fluorescence Microscopy,Immunofluorescence Microscopy,Microscopy, Immunofluorescence,Fluorescence Microscopies,Immunofluorescence Microscopies,Microscopies, Fluorescence,Microscopies, Immunofluorescence
D004330 Drosophila A genus of small, two-winged flies containing approximately 900 described species. These organisms are the most extensively studied of all genera from the standpoint of genetics and cytology. Fruit Fly, Drosophila,Drosophila Fruit Flies,Drosophila Fruit Fly,Drosophilas,Flies, Drosophila Fruit,Fly, Drosophila Fruit,Fruit Flies, Drosophila
D004625 Embryo, Nonmammalian The developmental entity of a fertilized egg (ZYGOTE) in animal species other than MAMMALS. For chickens, use CHICK EMBRYO. Embryonic Structures, Nonmammalian,Embryo, Non-Mammalian,Embryonic Structures, Non-Mammalian,Nonmammalian Embryo,Nonmammalian Embryo Structures,Nonmammalian Embryonic Structures,Embryo Structure, Nonmammalian,Embryo Structures, Nonmammalian,Embryo, Non Mammalian,Embryonic Structure, Non-Mammalian,Embryonic Structure, Nonmammalian,Embryonic Structures, Non Mammalian,Embryos, Non-Mammalian,Embryos, Nonmammalian,Non-Mammalian Embryo,Non-Mammalian Embryonic Structure,Non-Mammalian Embryonic Structures,Non-Mammalian Embryos,Nonmammalian Embryo Structure,Nonmammalian Embryonic Structure,Nonmammalian Embryos,Structure, Non-Mammalian Embryonic,Structure, Nonmammalian Embryo,Structure, Nonmammalian Embryonic,Structures, Non-Mammalian Embryonic,Structures, Nonmammalian Embryo,Structures, Nonmammalian Embryonic
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D047108 Embryonic Development Morphological and physiological development of EMBRYOS. Embryo Development,Embryogenesis,Postimplantation Embryo Development,Preimplantation Embryo Development,Embryonic Programming,Post-implantation Embryo Development,Postnidation Embryo Development,Postnidation Embryo Development, Animal,Pre-implantation Embryo Development,Prenidation Embryo Development, Animal,Development, Embryo,Development, Embryonic,Development, Postnidation Embryo,Embryo Development, Post-implantation,Embryo Development, Postimplantation,Embryo Development, Postnidation,Embryo Development, Pre-implantation,Embryo Development, Preimplantation,Embryonic Developments,Embryonic Programmings,Post implantation Embryo Development,Pre implantation Embryo Development
D021621 Imaging, Three-Dimensional The process of generating three-dimensional images by electronic, photographic, or other methods. For example, three-dimensional images can be generated by assembling multiple tomographic images with the aid of a computer, while photographic 3-D images (HOLOGRAPHY) can be made by exposing film to the interference pattern created when two laser light sources shine on an object. Computer-Assisted Three-Dimensional Imaging,Imaging, Three-Dimensional, Computer Assisted,3-D Image,3-D Imaging,Computer-Generated 3D Imaging,Three-Dimensional Image,Three-Dimensional Imaging, Computer Generated,3 D Image,3 D Imaging,3-D Images,3-D Imagings,3D Imaging, Computer-Generated,3D Imagings, Computer-Generated,Computer Assisted Three Dimensional Imaging,Computer Generated 3D Imaging,Computer-Assisted Three-Dimensional Imagings,Computer-Generated 3D Imagings,Image, 3-D,Image, Three-Dimensional,Images, 3-D,Images, Three-Dimensional,Imaging, 3-D,Imaging, Computer-Assisted Three-Dimensional,Imaging, Computer-Generated 3D,Imaging, Three Dimensional,Imagings, 3-D,Imagings, Computer-Assisted Three-Dimensional,Imagings, Computer-Generated 3D,Imagings, Three-Dimensional,Three Dimensional Image,Three Dimensional Imaging, Computer Generated,Three-Dimensional Images,Three-Dimensional Imaging,Three-Dimensional Imaging, Computer-Assisted,Three-Dimensional Imagings,Three-Dimensional Imagings, Computer-Assisted
D024462 Myosin Type II The subfamily of myosin proteins that are commonly found in muscle fibers. Myosin II is also involved a diverse array of cellular functions including cell division, transport within the GOLGI APPARATUS, and maintaining MICROVILLI structure. Myosin II

Related Publications

Mostafa Aakhte, and Hans-Arno J Müller
November 2015, Nature communications,
Mostafa Aakhte, and Hans-Arno J Müller
April 2013, Cell and tissue research,
Mostafa Aakhte, and Hans-Arno J Müller
March 2020, Methods (San Diego, Calif.),
Mostafa Aakhte, and Hans-Arno J Müller
June 2015, Nature methods,
Mostafa Aakhte, and Hans-Arno J Müller
August 2015, Biochemical and biophysical research communications,
Mostafa Aakhte, and Hans-Arno J Müller
November 2021, Nature biotechnology,
Mostafa Aakhte, and Hans-Arno J Müller
June 2016, Biomedical optics express,
Mostafa Aakhte, and Hans-Arno J Müller
August 2016, Journal of microscopy,
Mostafa Aakhte, and Hans-Arno J Müller
March 2024, Small methods,
Copied contents to your clipboard!