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Multiphoton Excitation Microscopy

Coherent recognizes that microscopists, neurologists, and biologists should not have to become laser experts to do their work. For this reason, we have designed and manufactured a wide range of lasers designed specifically for non-linear imaging. This way, MPE users can focus on their sample, not on the laser equipment.


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Interacting neural ensembles in orbitofrontal cortex for social and feeding behaviour

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Neurosciences

MerMAIDs: a family of metagenomically discovered marine anion-conducting and intensely desensitizing channelrhodopsins
MerMAIDs: a family of metagenomically discovered marine anion-conducting and intensely desensitizing channelrhodopsins
Channelrhodopsins (ChRs) are algal light-gated ion channels widely used as optogenetic tools for manipulating neuronal activity. ChRs desensitize under continuous bright-light illumination, resulting in a significant decline of photocurrents. Here we describe a metagenomically identified family of phylogenetically distinct anion-conducting ChRs (designated MerMAIDs). MerMAIDs almost completely desensitize during... - Johannes Oppermann, Paul Fischer, Arita Silapetere, Bernhard Liepe, Silvia Rodriguez-Rozada , José Flores-Uribe, Enrico Peter, Anke Keidel, Johannes Vierock, Joel Kaufmann, Matthias Broser, Meike Luck, Franz Bartl, Peter Hildebrandt, J. Simon Wiegert, Oded Béjà, Peter Hegemann & Jonas Wietek, 2019
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Interacting neural ensembles in orbitofrontal cortex for social and feeding behaviour
Categorically distinct basic drives (for example, for social versus feeding behaviour) can exert potent influences on each other; such interactions are likely to have important adaptive consequences (such as appropriate regulation of feeding in the context of social hierarchies) and can become maladaptive (such as in clinical settings involving anorexia). It is known that neural systems... - Joshua H. Jennings, Christina K. Kim, James H. Marshel, Misha Raffiee, Li Ye, Sean Quirin, Sally Pak, Charu Ramakrishnan & Karl Deisseroth, 2019
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Two-Photon Bidirectional Control and Imaging of Neuronal Excitability with High Spatial Resolution In Vivo
Sensory information is encoded within the brain in distributed spatiotemporal patterns of neuronal activity. Understanding how these patterns influence behavior requires a method to measure and to bidirectionally perturb with high spatial resolution the activity of the multiple neuronal cell types engaged in sensory processing. Here, we combined two-photon holography... - Hagai Har-Gil, Lior Golgher, Shai Israel, David Kain, Ori Cheshnovsky, Moshe Parnas, and Pablo Blinder, 2018
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Active learning of cortical connectivity from two-photon imaging data
Understanding how groups of neurons interact within a network is a fundamental question in system neuroscience. Instead of passively observing the ongoing activity of a network, we can typically perturb its activity, either by external sensory stimulation or directly via techniques such as two-photon optogenetics. A natural question is how to use such perturbations to identify the connectivity of the network... - Martin A. Bertran, Natalia L. Martinez, Ye Wang, David Dunson, Guillermo Sapiro, and Dario Ringach, 2018
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Fast varifocal two-photon microendoscope for imaging neuronal activity in the deep brain
Fluorescence microendoscopy is becoming a promising approach for deep brain imaging, but the current technology for visualizing neurons on a single focal plane limits the experimental efficiency and the pursuit of three-dimensional functional neural circuit architectures. Here we present a novel fast varifocal two-photon microendoscope system equipped with... - Masaaki Sato, Yuki Motegi, Shogo Yagi, Keiko Gengyo-Ando, Masamichi Ohkura, and Junichi Nakai, 2017
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Video rate volumetric Ca2+ imaging across cortex using seeded iterative demixing (SID) microscopy
Light-field microscopy (LFM) is a scalable approach for volumetric Ca2+ imaging with high volumetric acquisition rates (up to 100 Hz). Although the technology has enabled whole-brain Ca2+ imaging in semi-transparent specimens, tissue scattering has limited its application in the rodent brain. We introduce seeded iterative demixing (SID), a computational source-extraction technique that extends LFM to the mammalian cortex. - Tobias Nöbauer, Oliver Skocek, Alejandro J Pernía-Andrade, Lukas Weilguny, Francisca Martínez Traub, Maxim I Molodtsov, Alipasha Vaziri, 26 June 2017
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An optogenetic toolbox for unbiased discovery of functionally connected cells in neural circuits
Optical imaging approaches have revolutionized our ability to monitor neural network dynamics, but by themselves are unable to link a neuron’s activity to its functional connectivity. We present a versatile genetic toolbox, termed ‘Optobow’, for all-optical discovery of excitatory connections in vivo. - Dominique Förster, Marco Dal Maschio, Eva Laurell, Herwig Baier, 24 July 2017
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Large-field-of-view imaging by multi-pupil adaptive optics
Adaptive optics can correct for optical aberrations. We developed multi-pupil adaptive optics which enables simultaneous wavefront correction and expands the correction area to nine times that of conventional methods. - Jung-Hoon Park, Lingjie Kong, Yifeng Zhou, Meng Cui, 8 May 2017
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Genetically encoded calcium indicators
Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics - Looger et al. Front. Mol. Neurosci., 04 March 2013
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Improved deep two-photon calcium imaging
Two-photon laser scanning calcium imaging has emerged as a useful method for the exploration of neural function and structure at the cellular and subcellular level in vivo. - Birkner et al. Cell Calcium, 2016
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Extended view and illumination with time-division multiplexing
Here we analyze and demonstrate a method of using galvanometer mirrors to time-sequentially reposition multiple 3D holograms, both extending the field of view and increasing the total time-averaged two-photon signal. - Deisseroth et al. Optics Express 2015
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Simultaneous all-optical manipulation and recording of neural circuit activity with cellular resolution in vivo
We describe an all-optical strategy for simultaneously manipulating and recording the activity of multiple neurons with cellular resolution in vivo. - Packer, Hausser et al. Nature Methods 2014
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Two-photon optogenetics of dendritic spines and neural circuits
We demonstrate a two-photon optogenetic method that generates action potentials in neurons with single-cell precision, using the red-shifted opsin C1V1T. - Peterka, Yuste et al. Nature Methods 2012
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Cell Biology, Disease Studies and Intravital Imaging

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Multiphoton Microscopy Sets the Standard for Live-Cell Imaging
Tunable ultrafast lasers enable 3D imaging and research applications ranging from neuroscience to improved understanding of stem cell transplantation. - McCoy, Arrigoni. Biophotonics, April 2018
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Third harmonic generation microscopy of cells and tissue organization
The interaction of cells within their microenvironmental niche is fundamental to cell migration, positioning, growth, and differentiation in order to form and maintain complex tissue organization... - Friedl et al. Journal of Cell Science 2015
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In Vivo Imaging Reveals Extracellular VesicleMediated Phenocopying of Metastatic Behavior
Most cancer cells release heterogeneous populations of extracellular vesicles (EVs) containing proteins, lipids, and nucleic acids. In vitro experiments showed that EV uptake can lead to transfer of functional mRNA and altered cellular behavior. However, similar in vivo experiments remain challenging because ... - Jacco van Rheenen, Cell 2015
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Tissue-scale coordination of cellular behaviour promotes epidermal wound repair in live mice
Tissue repair is fundamental to our survival as tissues are challenged by recurrent damage. During mammalian skin repair, cells respond by... - Greco et al. Nature Cell biology, 2017
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Novel Techniques and Fluorescent Probe Development

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Scattering compensation by focus scanning holographic aberration probing (F-SHARP)
A long-standing goal in biomedical imaging, the control of light inside turbid media, requires knowledge of how the phase and amplitude of an illuminating wavefront are transformed as the electric field propagates inside a scattering sample onto a target plane. So far, it has proved challenging to... - Papadopolous et al. Nature Photonics 2016
Coherent
Continuous volumetric imaging via an optical phase-locked ultrasound lens
In vivo imaging at high spatiotemporal resolution is key to the understanding of complex biological systems. We integrated an optical phase-locked ultrasound lens into a two-photon fluorescence microscope and achieved... - Meng Cui et al.
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Single-wavelength two-photon excitation–stimulated emission depletion (SW2PE-STED) superresolution imaging
We developed a new class of two-photon excitation–stimulated emission depletion (2PE-STED) optical microscope. In this work, we show the opportunity to perform superresolved fluorescence imaging... - Diaspro et al. PNAS 2011v

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