scholarly journals Analytical Currents: Photocaged amino acids enable scientists to study phosphorylation in vivo | Two-photon excitation to stimulate and image neural circuits | Calorimetry for proteomics | Imaging carbon nanotubes in cells. | High-throughput, single-enzyme assay system | New IRMPD method for sequencing oligosaccharides

2008 ◽  
Vol 80 (1) ◽  
pp. 3-5
Keyword(s):  
Carbon Nanotubes ◽  
Amino Acids ◽  
High Throughput ◽  
Enzyme Assay ◽  
Neural Circuits ◽  
Assay System ◽  
Two Photon ◽  
Photon Excitation ◽  
Single Enzyme

scholarly journals Three-dimensional Two-Photon Optogenetics and Imaging of Neural Circuits in vivo

2017 ◽  
Author(s):  
Weijian Yang ◽  
Luis Carrillo-Reid ◽  
Yuki Bando ◽  
Darcy S. Peterka ◽  
Rafael Yuste
Keyword(s):  
Visual Cortex ◽  
Neural Activity ◽  
Calcium Imaging ◽  
Pyramidal Neurons ◽  
Neural Circuits ◽  
Three Dimensional ◽  
Two Photon ◽  
Cellular Resolution ◽  
Photon Excitation

We demonstrate a holographic system for simultaneous three-dimensional (3D) two-photon stimulation and imaging of neural activity in the mouse neocortex in vivo with cellular resolution. Dual two-photon excitation paths are implemented with independent 3D targeting for calcium imaging and precision optogenetics. We validate the usefulness of the microscope by photoactivating local pools of interneurons in awake mice visual cortex in 3D, which suppress the nearby pyramidal neurons’ response to visual stimuli.

scholarly journals Label-free concurrent 5-modal microscopy (Co5M) resolves unknown spatio-temporal processes in wound healing

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Markus Seeger ◽  
Christoph Dehner ◽  
Dominik Jüstel ◽  
Vasilis Ntziachristos
Keyword(s):  
Wound Healing ◽  
Label Free ◽  
Third Harmonic ◽  
Two Photon ◽  
Non Invasive ◽  
Photon Excitation ◽  
Free Mode ◽  
Spatio Temporal ◽  
And Function

AbstractThe non-invasive investigation of multiple biological processes remains a methodological challenge as it requires capturing different contrast mechanisms, usually not available with any single modality. Intravital microscopy has played a key role in dynamically studying biological morphology and function, but it is generally limited to resolving a small number of contrasts, typically generated by the use of transgenic labels, disturbing the biological system. We introduce concurrent 5-modal microscopy (Co5M), illustrating a new concept for label-free in vivo observations by simultaneously capturing optoacoustic, two-photon excitation fluorescence, second and third harmonic generation, and brightfield contrast. We apply Co5M to non-invasively visualize multiple wound healing biomarkers and quantitatively monitor a number of processes and features, including longitudinal changes in wound shape, microvascular and collagen density, vessel size and fractality, and the plasticity of sebaceous glands. Analysis of these parameters offers unique insights into the interplay of wound closure, vasodilation, angiogenesis, skin contracture, and epithelial reformation in space and time, inaccessible by other methods. Co5M challenges the conventional concept of biological observation by yielding multiple simultaneous parameters of pathophysiological processes in a label-free mode.

scholarly journals Conformationally rigid pyrazoloquinazoline α-amino acids: one- and two-photon induced fluorescence

2020 ◽  
Vol 56 (12) ◽  
pp. 1887-1890 ◽  
Author(s):  
Jonathan D. Bell ◽  
Alexander H. Harkiss ◽  
David Nobis ◽  
Eilidh Malcolm ◽  
Astrid Knuhtsen ◽  
...  
Keyword(s):  
Amino Acids ◽  
Fluorescent Probes ◽  
Ring System ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

Conformationally rigid unnatural α-amino acids bearing a pyrazoloquinazoline ring system that are amenable to both one- and two-photon excitation have been developed as new fluorescent probes.

In vivo label-free two-photon excitation autofluorescence microscopy of microvasculature using a 520 nm femtosecond fiber laser

2020 ◽  
Vol 45 (10) ◽  
pp. 2704
Author(s):  
Ting Wu ◽  
Jiuling Liao ◽  
Jia Yu ◽  
Yufeng Gao ◽  
Hui Li ◽  
...  
Keyword(s):  
Fiber Laser ◽  
Label Free ◽  
Two Photon ◽  
Photon Excitation ◽  
Femtosecond Fiber Laser ◽  
Two Photon Excitation

Intravital bone imaging by two-photon excitation microscopy to identify osteocytic osteolysis in vivo

Bone ◽  
2015 ◽  
Vol 74 ◽  
pp. 134-139 ◽  
Author(s):  
Hiroshige Sano ◽  
Junichi Kikuta ◽  
Masayuki Furuya ◽  
Naoki Kondo ◽  
Naoto Endo ◽  
...  
Keyword(s):  
Bone Imaging ◽  
Two Photon ◽  
Photon Excitation ◽  
Osteocytic Osteolysis ◽  
Two Photon Excitation

scholarly journals A novel method for observing proteins in vivo using a small fluorescent label and multiphoton imaging

2005 ◽  
Vol 390 (3) ◽  
pp. 787-790 ◽  
Author(s):  
Stanley W. Botchway ◽  
Ignasi Barba ◽  
Randolf Jordan ◽  
Rebecca Harmston ◽  
Peter M. Haggie ◽  
...  
Keyword(s):  
Fluorescence Detection ◽  
Fluorescence Emission ◽  
Yeast Cells ◽  
Fluorescent Label ◽  
Multiphoton Imaging ◽  
Two Photon ◽  
Photon Excitation ◽  
Novel Method ◽  
Two Photon Excitation

A novel method for the fluorescence detection of proteins in cells is described in the present study. Proteins are labelled by the selective biosynthetic incorporation of 5-hydroxytryptophan and the label is detected via selective two-photon excitation of the hydroxyindole and detection of its fluorescence emission at 340 nm. The method is demonstrated in this paper with images of a labelled protein in yeast cells.

scholarly journals Dual-color volumetric imaging of neural activity of cortical columns

2018 ◽  
Author(s):  
Shuting Han ◽  
Weijian Yang ◽  
Rafael Yuste
Keyword(s):  
Neural Activity ◽  
High Speed ◽  
Neural Circuits ◽  
Emergent Properties ◽  
Spatiotemporal Resolution ◽  
Population Activity ◽  
Volumetric Imaging ◽  
Two Photon ◽  
Dual Color

To capture the emergent properties of neural circuits, high-speed volumetric imaging of neural activity at cellular resolution is desirable. But while conventional two-photon calcium imaging is a powerful tool to study population activity in vivo, it is restrained to two-dimensional planes. Expanding it to 3D while maintaining high spatiotemporal resolution appears necessary. Here, we developed a two-photon microscope with dual-color laser excitation that can image neural activity in a 3D volume. We imaged the neuronal activity of primary visual cortex from awake mice, spanning from L2 to L5 with 10 planes, at a rate of 10 vol/sec, and demonstrated volumetric imaging of L1 long-range PFC projections and L2/3 somatas. Using this method, we map visually-evoked neuronal ensembles in 3D, finding a lack of columnar structure in orientation responses and revealing functional correlations between cortical layers which differ from trial to trial and are missed in sequential imaging. We also reveal functional interactions between presynaptic L1 axons and postsynaptic L2/3 neurons. Volumetric two-photon imaging appears an ideal method for functional connectomics of neural circuits.

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