Light sheet microscopy for fast functional imaging of 3D neuronal cultures in hydrogels

2020 ◽  
Author(s):  
Gustavo Castro-Olvera ◽  
Jorge Madrid-Wolff ◽  
Omar E. Olarte ◽  
Estefanía Estévez-Priego ◽  
Adriaan A. Ludl ◽  
...  
Keyword(s):  
Functional Imaging ◽  
Light Sheet ◽  
Neuronal Cultures ◽  
Light Sheet Microscopy

scholarly journals CaMKIIα Expressing Neurons to Report Activity-Related Endogenous Hypoxia upon Motor-Cognitive Challenge

2021 ◽  
Vol 22 (6) ◽  
pp. 3164
Author(s):  
Umer Javed Butt ◽  
Imam Hassouna ◽  
Laura Fernandez Garcia-Agudo ◽  
Agnes A. Steixner-Kumar ◽  
Constanze Depp ◽  
...  
Keyword(s):  
Pyramidal Neurons ◽  
Cognitive Task ◽  
Light Sheet ◽  
Brain Maturation ◽  
Neuronal Cultures ◽  
Behavioral Challenges ◽  
Light Sheet Microscopy ◽  
Improved Performance ◽  
Cag Promoter

We previously introduced the brain erythropoietin (EPO) circle as a model to explain the adaptive ‘brain hardware upgrade’ and enhanced performance. In this fundamental circle, brain cells, challenged by motor-cognitive tasks, experience functional hypoxia, triggering the expression of EPO among other genes. We attested hypoxic cells by a transgenic reporter approach under the ubiquitous CAG promoter, with Hif-1α oxygen-dependent degradation-domain (ODD) fused to CreERT2-recombinase. To specifically focus on the functional hypoxia of excitatory pyramidal neurons, here, we generated CaMKIIα-CreERT2-ODD::R26R-tdTomato mice. Behavioral challenges, light-sheet microscopy, immunohistochemistry, single-cell mRNA-seq, and neuronal cultures under normoxia or hypoxia served to portray these mice. Upon complex running wheel performance as the motor-cognitive task, a distinct increase in functional hypoxic neurons was assessed immunohistochemically and confirmed three-dimensionally. In contrast, fear conditioning as hippocampal stimulus was likely too short-lived to provoke neuronal hypoxia. Transcriptome data of hippocampus under normoxia versus inspiratory hypoxia revealed increases in CA1 CaMKIIα-neurons with an immature signature, characterized by the expression of Dcx, Tbr1, CaMKIIα, Tle4, and Zbtb20, and consistent with accelerated differentiation. The hypoxia reporter response was reproduced in vitro upon neuronal maturation. To conclude, task-associated activity triggers neuronal functional hypoxia as a local and brain-wide reaction mediating adaptive neuroplasticity. Hypoxia-induced genes such as EPO drive neuronal differentiation, brain maturation, and improved performance.

Increasing sensitivity and accuracy of brain-wide quantitative studies in light-sheet microscopy

2017 ◽  
Author(s):  
Caroline M. Müllenbroich ◽  
Ludovico Silvestri ◽  
Lapo Turrini ◽  
Tommaso Alterini ◽  
Antonino P. Di Giovanna ◽  
...  
Keyword(s):  
Functional Imaging ◽  
Fold Increase ◽  
Structural Data ◽  
Light Sheet ◽  
High Fidelity ◽  
Random Errors ◽  
Bessel Beams ◽  
Light Sheet Microscopy ◽  
Quantitative Studies ◽  
Increase In Accuracy

AbstractLight-sheet microscopy (LSM) has proven a useful tool in neuroscience to image whole brains with high frame rates at cellular resolution. LSM is employed either in combination with tissue clearing to reconstruct the cyto-architecture over the entire mouse brain or with intrinsically transparent samples like zebrafish larvae for functional imaging. Inherently to LSM, however, residual opaque objects cause stripe artifacts, which obscure features of interest and, during functional imaging, modulate fluorescence variations related to neuronal activity. Here, we report how Bessel beams reduce streaking artifacts and produce high-fidelity structural data. Furthermore, using Bessel beams, we demonstrate a fivefold increase in sensitivity to calcium transients and a 20 fold increase in accuracy in the detection of activity correlations in functional imaging. Our results demonstrate the contamination of data by systematic and random errors through Gaussian illumination and furthermore quantify the increase in fidelity of such data when using Bessel beams.

Whole-brain functional imaging with two-photon light-sheet microscopy

2015 ◽  
Vol 12 (5) ◽  
pp. 379-380 ◽  
Author(s):  
Sébastien Wolf ◽  
Willy Supatto ◽  
Georges Debrégeas ◽  
Pierre Mahou ◽  
Sergei G Kruglik ◽  
...  
Keyword(s):  
Functional Imaging ◽  
Light Sheet ◽  
Whole Brain ◽  
Two Photon ◽  
Light Sheet Microscopy

Functional Imaging with Light-Sheet Microscopy

2020 ◽  
pp. 21-54
Author(s):  
Raghav K. Chhetri ◽  
Philipp J. Keller
Keyword(s):  
Functional Imaging ◽  
Light Sheet ◽  
Light Sheet Microscopy

scholarly journals Two-photon light-sheet microscopy for high-speed whole-brain functional imaging of zebrafish neuronal physiology and pathology

2020 ◽  
Author(s):  
Giuseppe de Vito ◽  
Lapo Turrini ◽  
Chiara Fornetto ◽  
Pietro Ricci ◽  
Caroline Müllenbroich ◽  
...  
Keyword(s):  
Functional Imaging ◽  
High Speed ◽  
Light Sheet ◽  
Whole Brain ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
Neuronal Physiology

Whole-brain functional imaging at cellular resolution using light-sheet microscopy

2013 ◽  
Vol 10 (5) ◽  
pp. 413-420 ◽  
Author(s):  
Misha B Ahrens ◽  
Michael B Orger ◽  
Drew N Robson ◽  
Jennifer M Li ◽  
Philipp J Keller
Keyword(s):  
Functional Imaging ◽  
Light Sheet ◽  
Whole Brain ◽  
Light Sheet Microscopy ◽  
Cellular Resolution
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