scholarly journals Effects of excitation light polarization on fluorescence emission in two-photon light-sheet microscopy

2020 ◽  
Vol 11 (8) ◽  
pp. 4651
Author(s):  
Giuseppe de Vito ◽  
Pietro Ricci ◽  
Lapo Turrini ◽  
Vladislav Gavryusev ◽  
Caroline Müllenbroich ◽  
...  
Keyword(s):  
Fluorescence Emission ◽  
Light Sheet ◽  
Light Polarization ◽  
Excitation Light ◽  
Two Photon ◽  
Light Sheet Microscopy

scholarly journals Characterization of Scattering Effects in Phantom Samples using Single and Two-Photon Excitation Light Sheet Microscopy

2012 ◽  
Vol 102 (3) ◽  
pp. 195a-196a
Author(s):  
Zeno Lavagnino ◽  
Francesca Cella Zanacchi ◽  
Emiliano Ronzitti ◽  
Ivan Coto Hernandez ◽  
Alberto Diaspro
Keyword(s):  
Light Sheet ◽  
Excitation Light ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
Photon Excitation ◽  
Scattering Effects ◽  
Two Photon Excitation

scholarly journals Fast, multicolor 3-D imaging of brain organoids with a new single-objective two-photon virtual light-sheet microscope

2018 ◽  
Author(s):  
Irina Rakotoson ◽  
Brigitte Delhomme ◽  
Philippe Djian ◽  
Andreas Deeg ◽  
Maia Brunstein ◽  
...  
Keyword(s):  
Spatial Resolution ◽  
Three Dimensional ◽  
Numerical Aperture ◽  
Light Sheet ◽  
Confocal Microscope ◽  
Large Field ◽  
Excitation Light ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
Functional Features

ABSTRACTHuman inducible pluripotent stem cells (hiPSCs) hold a large potential for disease modeling. hiPSC-derived human astrocyte and neuronal cultures permit investigations of neural signaling pathways with subcellular resolution. Combinatorial cultures, and three-dimensional (3-D) embryonic bodies enlarge the scope of investigations to multi-cellular phenomena. A the highest level of complexity, brain organoids that – in many aspects – recapitulate anatomical and functional features of the developing brain permit the study of developmental and morphological aspects of human disease. An ideal microscope for 3-D tissue imaging at these different scales would combine features from both confocal laser-scanning and light-sheet microscopes: a micrometric optical sectioning capacity and sub-micrometric spatial resolution, a large field of view and high frame rate, and a low degree of invasiveness, i.e., ideally, a better photon efficiency than that of a confocal microscope. In the present work, we describe such an instrument that belongs to the class of two-photon (2P) light-sheet microsocpes. Its particularity is that – unlike existing two- or three-lens designs – it is using a single, low-magnification, high-numerical aperture objective for the generation and scanning of a virtual light sheet. The microscope builds on a modified Nipkow-Petran spinning-disk scheme for achieving wide-field excitation. However, unlike the common Yokogawa design that uses a tandem disk, our concept combines micro lenses, dichroic mirrors and detection pinholes on a single disk. This design, advantageous for 2P excitation circumvents problems arising with the tandem disk from the large wavelength-difference between the infrared excitation light and visible fluorescence. 2P fluorescence excited in by the light sheet is collected by the same objective and imaged onto a fast sCMOS camera. We demonstrate three-dimensional imaging of TO-PRO3-stained embryonic bodies and of brain organoids, under control conditions and after rapid (partial) transparisation with triethanolamine and /ormamide (RTF) and compare the performance of our instrument to that of a confocal microscope having a similar numerical aperture. 2P-virtual light-sheet microscopy permits one order of magnitude faster imaging, affords less photobleaching and permits better depth penetration than a confocal microscope with similar spatial resolution.

scholarly journals Wide field intravital imaging by two-photon-excitation digital-scanned light-sheet microscopy (2p-DSLM) with a high-pulse energy laser

2014 ◽  
Vol 5 (10) ◽  
pp. 3311 ◽  
Author(s):  
Atsushi Maruyama ◽  
Yusuke Oshima ◽  
Hiroko Kajiura-Kobayashi ◽  
Shigenori Nonaka ◽  
Takeshi Imamura ◽  
...  
Keyword(s):  
Pulse Energy ◽  
Light Sheet ◽  
Intravital Imaging ◽  
Wide Field ◽  
High Pulse Energy ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
Photon Excitation ◽  
High Pulse ◽  
Energy Laser

scholarly journals Light-sheet microscopy in thick media using scanned Bessel beams and two-photon fluorescence excitation

2013 ◽  
Vol 21 (11) ◽  
pp. 13824 ◽  
Author(s):  
Florian O. Fahrbach ◽  
Vasily Gurchenkov ◽  
Kevin Alessandri ◽  
Pierre Nassoy ◽  
Alexander Rohrbach
Keyword(s):  
Light Sheet ◽  
Bessel Beams ◽  
Fluorescence Excitation ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

Two-photon three-axis digital scanned light-sheet microscopy (2P3A-DSLM)

CLEO: 2014 ◽  
2014 ◽  
Author(s):  
Weijian Zong ◽  
Xuanyang Chen ◽  
Jia Zhao ◽  
Yunfeng Zhang ◽  
Ming Fan ◽  
...  
Keyword(s):  
Light Sheet ◽  
Two Photon ◽  
Light Sheet Microscopy

scholarly journals Multiview Deconvolution Two-photon Laser Scanning Microscopy

2020 ◽  
Author(s):  
Dimitrios Kapsokalyvas ◽  
Rodrigo Rosas ◽  
Rob Janssen ◽  
Jo Vanoevelen ◽  
Martin Strauch ◽  
...  
Keyword(s):  
Second Harmonic ◽  
Light Sheet ◽  
Laser Scanning Microscopy ◽  
Three Dimensions ◽  
Scanning Microscopy ◽  
Fluorescence Signal ◽  
Microscopy Imaging ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
3D Image Reconstruction

Abstract Imaging in three dimensions is necessary for thick tissues and small organisms. This is possible with tomographic optical microscopy techniques such as confocal, two-photon and light sheet microscopy. All these techniques suffer from anisotropic resolution and limited penetration depth. In the past, Multiview microscopy - imaging the sample from different angles followed by 3D image reconstruction - was developed to address this issue for light sheet microscopy based on fluorescence signal. In this study we applied this methodology to accomplish Multiview imaging with two-photon microscopy based on fluorescence and additionally second harmonic signal from myosin and collagen. It was shown that isotropic resolution was achieved, the entirety of the sample was visualized, and interference artifacts were suppressed allowing clear visualization of collagen fibrils and myofibrils. This method can be applied to any scanning microscopy technique without microscope modifications. It can be used for imaging tissue and whole mount small organisms such as heart tissue, and zebrafish larva in 3D, label-free or stained, with at least 3-fold axial resolution improvement which can be significant for the accurate quantification of small 3D structures.

scholarly journals Imaging Proteins, Cells, and Tissues Dynamics during Embryogenesis with Two-Photon Light-Sheet Microscopy

2013 ◽  
Vol 104 (2) ◽  
pp. 337a
Author(s):  
Thai V. Truong ◽  
Daniel B. Holland ◽  
Vikas Trivedi ◽  
Scott E. Fraser
Keyword(s):  
Light Sheet ◽  
Two Photon ◽  
Light Sheet Microscopy

scholarly journals Multicolor two-photon light-sheet microscopy

2014 ◽  
Vol 11 (6) ◽  
pp. 600-601 ◽  
Author(s):  
Pierre Mahou ◽  
Julien Vermot ◽  
Emmanuel Beaurepaire ◽  
Willy Supatto
Keyword(s):  
Light Sheet ◽  
Two Photon ◽  
Light Sheet Microscopy

scholarly journals Wide field light-sheet microscopy with lens-axicon controlled two-photon Bessel beam illumination

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Sota Takanezawa ◽  
Takashi Saitou ◽  
Takeshi Imamura
Keyword(s):  
High Speed ◽  
Bessel Beam ◽  
Light Sheet ◽  
Time Lapse ◽  
Whole Body ◽  
Wide Field ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
Photon Excitation ◽  
Two Photon Excitation

AbstractTwo-photon excitation can lower phototoxicity and improve penetration depth, but its narrow excitation range restricts its applications in light-sheet microscopy. Here, we propose simple illumination optics, a lens-axicon triplet composed of an axicon and two convex lenses, to generate longer extent Bessel beams. This unit can stretch the beam full width at half maximum of 600–1000 μm with less than a 4-μm waist when using a 10× illumination lens. A two-photon excitation digital scanned light-sheet microscope possessing this range of field of view and ~2–3-μm axial resolution is constructed and used to analyze the cellular dynamics over the whole body of medaka fish. We demonstrate long-term time-lapse observations over several days and high-speed recording with ~3 mm3 volume per 4 s of the embryos. Our system is minimal and suppresses laser power loss, which can broaden applications of two-photon excitation in light-sheet microscopy.

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