scholarly journals Optical clearing of living brains with MAGICAL to extend in vivo imaging

2019 ◽  
Author(s):  
Kouichirou Iijima ◽  
Takuto Oshima ◽  
Ryosuke Kawakami ◽  
Tomomi Nemoto
Keyword(s):  
Fluorescence Emission ◽  
Cortical Layer ◽  
Fluorescence Emission Spectrum ◽  
Spatiotemporal Resolution ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Brain Functions ◽  
Hippocampal Ca1 ◽  
Layer V

AbstractTo understand brain functions, it is important to observe directly how multiple neural circuits are performing in living brains. However, due to tissue opaqueness, observable depth and spatiotemporal resolution are severely degraded in vivo. Here, we propose an optical brain clearing method for in vivo fluorescence microscopy, termed MAGICAL (Magical Additive Glycerol Improves Clear Alive Luminance). MAGICAL enabled two-photon microscopy to capture vivid images with fast speed, at cortical layer V and hippocampal CA1 in vivo. Moreover, MAGICAL promoted conventional confocal microscopy to visualize finer neuronal structures including synaptic boutons and spines in unprecedented deep regions, without intensive illumination leading to phototoxic effects. Fluorescence Emission Spectrum Transmissive Analysis (FESTA) showed that MAGICAL improved in vivo transmittance of shorter wavelength light, which is vulnerable to optical scattering thus unsuited for in vivo microscopy. These results suggest that MAGICAL would transparentize living brains via scattering reduction.

scholarly journals In Vivo Imaging of Neocortical and Hippocampal CA1 Neurons by Two-photon Microscopy

2014 ◽  
Vol 54 (1) ◽  
pp. 035-038
Author(s):  
Ryosuke KAWAKAMI ◽  
Terumasa HIBI ◽  
Tomomi NEMOTO
Keyword(s):  
In Vivo Imaging ◽  
Ca1 Neurons ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Hippocampal Ca1 Neurons ◽  
Hippocampal Ca1

In vivo imaging of liver injury and regeneration by functional two-photon microscopy

2016 ◽  
Vol 54 (12) ◽  
pp. 1343-1404
Author(s):  
A Ghallab ◽  
R Reif ◽  
R Hassan ◽  
AS Seddek ◽  
JG Hengstler
Keyword(s):  
Liver Injury ◽  
In Vivo Imaging ◽  
Two Photon Microscopy ◽  
Two Photon

scholarly journals High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiang Lan Fan ◽  
Jose A. Rivera ◽  
Wei Sun ◽  
John Peterson ◽  
Henry Haeberle ◽  
...  
Keyword(s):  
Blood Flow ◽  
High Speed ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Fluorescence Signal ◽  
Imaging Method ◽  
Spatiotemporal Resolution ◽  
Two Photon

AbstractUnderstanding the structure and function of vasculature in the brain requires us to monitor distributed hemodynamics at high spatial and temporal resolution in three-dimensional (3D) volumes in vivo. Currently, a volumetric vasculature imaging method with sub-capillary spatial resolution and blood flow-resolving speed is lacking. Here, using two-photon laser scanning microscopy (TPLSM) with an axially extended Bessel focus, we capture volumetric hemodynamics in the awake mouse brain at a spatiotemporal resolution sufficient for measuring capillary size and blood flow. With Bessel TPLSM, the fluorescence signal of a vessel becomes proportional to its size, which enables convenient intensity-based analysis of vessel dilation and constriction dynamics in large volumes. We observe entrainment of vasodilation and vasoconstriction with pupil diameter and measure 3D blood flow at 99 volumes/second. Demonstrating high-throughput monitoring of hemodynamics in the awake brain, we expect Bessel TPLSM to make broad impacts on neurovasculature research.

Sensory Transmission is Bi-directionally Modulated by Astrocytic Ca2+ in Barrel Cortex of Behaving Mice

2021 ◽  
Author(s):  
Simeng Gu ◽  
Wei Wang ◽  
Kuan Zhang ◽  
Rou Feng ◽  
Naling Li ◽  
...  
Keyword(s):  
Sensory Input ◽  
Barrel Cortex ◽  
Synaptic Function ◽  
Metabolic Clearance ◽  
Sleep State ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
Sensory Transmission

Abstract Different effects of astrocyte during sleep and awake have been extensively studied, especially for metabolic clearance by the glymphatic system, which works during sleep and stops working during waking states. However, how astrocytes contribute to modulation of sensory transmission during sleep and awake animals remain largely unknown. Recent advances in genetically encoded Ca2+ indicators have provided a wealth of information on astrocytic Ca2+, especially in their fine perisynaptic processes, where astrocytic Ca2+ most likely affects the synaptic function. Here we use two-photon microscopy to image astrocytic Ca2+ signaling in freely moving mice trained to run on a wheel in combination with in vivo whole-cell recordings to evaluate the role of astrocytic Ca2+ signaling in different behavior states. We found that there are two kinds of astrocytic Ca2+ signaling: a small long-lasting Ca2+ increase during sleep state and a sharp widespread but short-long-lasting Ca2+ spike when the animal was awake (fluorescence increases were 23.2 ± 14.4% for whisker stimulation at sleep state, compared with 73.3 ± 11.7% for at awake state, paired t-test, p < 0.01). The small Ca2+ transients decreased extracellular K+, hyperpolarized the neurons, and suppressed sensory transmission; while the large Ca2+ wave enhanced sensory input, contributing to reliable sensory transmission in aroused states. Locus coeruleus activation works as a switch between these two kinds of astrocytic Ca2+ elevation. Thus, we show that cortical astrocytes play an important role in processing of sensory input. These two types of events appear to have different pharmacological sources and may play a different role in facilitating the efficacy of sensory transmission.

Quantum Dots assisted and NIR-II Emissive In Vivo Two-photon microscopy

2021 ◽  
Author(s):  
Huwei Ni ◽  
Yalun Wang ◽  
Tao Tang ◽  
Wenbin Yu ◽  
Dongyu Li ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Two Photon Microscopy ◽  
Two Photon

Video-rate two-photon microscopy of cortical hemodynamics in-vivo

2006 ◽  
Author(s):  
Matthew Bouchard ◽  
Svetlana Ruvinskya ◽  
David A. Boas ◽  
Elizabeth M. C. Hillman
Keyword(s):  
Two Photon Microscopy ◽  
Two Photon

scholarly journals Impact of Body Site, Age, and Gender on the Collagen/Elastin Index by Noninvasive in vivo Vertical Two-Photon Microscopy

2017 ◽  
Vol 30 (5) ◽  
pp. 260-267 ◽  
Author(s):  
Carolin Czekalla ◽  
Karl-Heinz Schönborn ◽  
Nadine Döge ◽  
Sora Jung ◽  
Maxim E. Darvin ◽  
...  
Keyword(s):  
Body Site ◽  
Age And Gender ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
And Gender

scholarly journals P02.08 Visualizing tumor cell - lymphocyte interactions in the brain metastatic cascade using in vivo two photon microscopy

2018 ◽  
Vol 20 (suppl_3) ◽  
pp. iii273-iii273
Author(s):  
M Piechutta ◽  
A S Berghoff ◽  
M A Karreman ◽  
K Gunkel ◽  
W Wick ◽  
...  
Keyword(s):  
Tumor Cell ◽  
Metastatic Cascade ◽  
Two Photon Microscopy ◽  
Two Photon ◽  
The Brain

scholarly journals Hyperspectral in vivo two-photon microscopy of intrinsic contrast

2008 ◽  
Vol 33 (18) ◽  
pp. 2164 ◽  
Author(s):  
Andrew J. Radosevich ◽  
Matthew B. Bouchard ◽  
Sean A. Burgess ◽  
Brenda R. Chen ◽  
Elizabeth M. C. Hillman
Keyword(s):  
Two Photon Microscopy ◽  
Two Photon
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