3:30pm - 4:00pmInvitedID: 513
/ TOM3 S03: 1
TOM 3 BioPhotonics
Volumetric one-photon UVA hyperspectral light sheet imaging in mouse pre-implantation embryos
Josephine Morizet1, Darren Chow2, Philip Wijesinghe1, Erik Schartner3, George Dwapanyin1, Kylie Dunning2, Kishan Dholakia1,3
1SUPA , School of Physics and Astronomy, University of St Andrews, North Haugh, St Andrews, Fife, United Kingdom; 2Robinson Research Institute, School of Biomedicine, The University of Adelaide, Adelaide, Australia; 3Centre of Light for Life and School of Biological Sciences, The University of Adelaide, Adelaide, 5005, Australia
UVA one-photon hyperspectral light sheet imaging was performed here to capture and reconstruct three dimensional metabolic maps of mouse embryos during development. We explored numerically and experimentally the advantages of phasor-based hyperspectral approach for detecting embryo metabolism with a single wavelength excitation compared to the conventional detection approach using bandpass filters.
4:00pm - 4:15pmID: 407
/ TOM3 S03: 2
TOM 3 BioPhotonics
Structured-light-sheet imaging on a chip
Petra Paiè1, Gianmaria Calisesi1, Alessia Candeo1, Andrea Comi2, Federico Sala2, Francesco Ceccarelli2, Ada De Luigi3, Pietro Veglianese3, Korbinian Muhlberger4, Michael Fokine4, Gianluca Valentini1, Roberto Osellame2, Mark Neil5, Andrea Bassi1, Francesca Bragheri2
1Dipartimento di Fisica, Politecnico di Milano, Piazza Leonardo da Vinci, 32, Milano, 20133, Italy.; 2Istituto di Fotonica e Nanotecnologie, Consiglio Nazionale delle Ricerche, Piazza Leonardo da Vinci, 32, Milano, 20133, Italy.; 3Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Via Mario Negri 2, Milano, 20156, Italy.; 4Department of Applied Physics, KTH Royal Institute of Technology, Roslagstullsbacken 21, Stockholm, 11421, Sweden.; 5Physics Department, Imperial College London, Prince Consort Road, London, SW7 2BB, UK.
In this work we present a microscope on a chip for automatic structured light sheet imaging of single cells. This integrated platform has been fabricated by femtosecond laser micromachining in glass substrates and it encompasses integrated optical elements such as waveguides, lenses and beam splitters as well as a microfluidic channel for sample delivery. Processing fluorescent cells, we have proven enhanced image resolution, automatic and continuous imaging as well as device ease of use
4:15pm - 4:30pmID: 405
/ TOM3 S03: 3
TOM 3 BioPhotonics
Imaging of calcium gradient oscillations in plant root hairs by light sheet fluorescence microscopy
Giorgia Tortora1, Stefano Buratti2, Matteo Grenzi2, Alex Costa2, Andrea Bassi1, Alessia Candeo1
1Politecnico di Milano, Italy; 2Università degli Studi di Milano, Italy
Root hairs are a delicate single-cell system whose growth is regulated by a fine mechanism characterised by the presence of a tip-high Ca2+ gradient that shows regular oscillations in growing root hairs. We show a method based on the use of Light sheet fluorescence microscopy (LSFM) which allows the quasi-physiological analysis of Arabidopsis thaliana plant roots hairs with excellent spatial and temporal resolution over a wide field of view. We show how the healthy growing root hairs are linked to precise oscillations and how a disruption of this mechanism can be associated to specific genes.
4:30pm - 4:45pmID: 193
/ TOM3 S03: 4
TOM 3 BioPhotonics
Diffuser-based fiber endoscopy for single-shot 3D fluorescence imaging
Tom Glosemeyer, Julian Lich, Robert Kuschmierz, Jürgen Czarske
TU Dresden, Germany
Minimally invasive endoscopy using coherent fiber bundles shows great potential for numerous applications in biomedical imaging. With a diffuser on the distal side of the fiber bundle and computational image recovery, single-shot 3D imaging is possible by encoding the image volume into 2D speckle patterns. In comparison to equivalent lens systems, a higher space-bandwidth product can be achieved. However, decoding the image with iterative algorithms is time-consuming. Thus, we propose utilizing a neural network for fast 2D and 3D image reconstruction at video rate. In this work, single-shot 3D fluorescence imaging with an ultra-thin endoscope is demonstrated, enabling applications like calcium imaging for in vivo brain diagnostics at cellular resolution.
4:45pm - 5:00pmID: 380
/ TOM3 S03: 5
TOM 3 BioPhotonics
Smart-scanning strategy for 3D imaging and laser manipulation of embryonic structures
Faris Abouakil, Huicheng Meng, Dmitry Nuzdhin, Miguel Sisson, Galland Frédéric, Loïc Le Goff
Aix Marseille Univ, CNRS, Centrale Marseille, Institut Fresnel, Turing Center for Living Systems, Marseille, France
Scanning fluorescence microscopes can image large tissues at the diffraction-limit. Scanning systems are also used for manipulation such as laser severing or optogenetic activation.
The conventional approach to image tissues with a scanning microscope involves scanning the entire bounding box enclosing the tissue, plane by plane, which is time-consuming and results in a high light dose.
We have developed a “smart” microscope that adapts its scanning scheme to the morphology of embryonic cell sheets, with no prior knowledge of the structure of interest. The surface of the tissue is first delineated from the acquisition of a very fractional scan of sample-space, using a robust estimation strategy. Subsequent high-resolution imaging can then be restricted to this targeted structure of interest, yielding a ~20-fold reduction in the scan path and, thus, an equivalent reduction in light dose and increase in speed. Additional scan path reduction can also be obtained using a propagative scanning approach. The resulting reduction in light dose (order of magnitude) is highly beneficial in terms of photobleaching.
We demonstrate the efficacy of our smart-scanning technique imaging Drosophila embryos and demonstrate how it can also be employed in a smart dissection scheme to make precise cuts on large non-planar tissues.
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