scholarly journals Balancing signal and photoperturbation in multiphoton light-sheet microscopy by optimizing laser pulse frequency

2020 ◽  
Author(s):  
Vincent Maioli ◽  
Antoine Boniface ◽  
Pierre Mahou ◽  
Júlia Ferrer Ortas ◽  
Lamiae Abdeladim ◽  
...  
Keyword(s):  
Laser Pulse ◽  
Multiphoton Microscopy ◽  
Light Sheet ◽  
Pulse Frequency ◽  
Heart Beat ◽  
Research Field ◽  
Tissue Imaging ◽  
Power Pulse ◽  
Light Sheet Microscopy

AbstractImproving the imaging speed of multiphoton microscopy is an active research field. Among recent strategies, light-sheet illumination holds distinctive advantages for achieving fast imaging in vivo. However, photoperturbation in multiphoton light-sheet microscopy remains poorly investigated. We show here that the heart beat rate of zebrafish embryos is a sensitive probe of linear and nonlinear photoperturbations. By analyzing its behavior with respect to laser power, pulse frequency and wavelength, we derive guidelines to balance signal and photoperturbation. We then demonstrate one order-of-magnitude signal enhancement over previous implementations by optimizing the laser pulse frequency. These results open new opportunities for fast live tissue imaging.

Multiphoton Light-sheet Microscopy at Optimal Pulse Frequency for Fast In Vivo Imaging

2020 ◽  
Author(s):  
Vincent Maioli ◽  
Antoine Boniface ◽  
Pierre Mahou ◽  
Júlia Ferrer Ortas ◽  
Lamiae Abdeladim ◽  
...  
Keyword(s):  
In Vivo Imaging ◽  
Light Sheet ◽  
Pulse Frequency ◽  
Light Sheet Microscopy

Fast in vivo imaging using multiphoton light-sheet microscopy at optimal pulse frequency

2021 ◽  
Author(s):  
Vincent Maioli ◽  
Antoine Boniface ◽  
Pierre Mahou ◽  
Júlia Ferrer Ortas ◽  
Lamiae Abdeladim ◽  
...  
Keyword(s):  
In Vivo Imaging ◽  
Light Sheet ◽  
Pulse Frequency ◽  
Light Sheet Microscopy

scholarly journals Fast in vivo multiphoton light-sheet microscopy with optimal pulse frequency

2020 ◽  
Vol 11 (10) ◽  
pp. 6012 ◽  
Author(s):  
Vincent Maioli ◽  
Antoine Boniface ◽  
Pierre Mahou ◽  
Júlia Ferrer Ortas ◽  
Lamiae Abdeladim ◽  
...  
Keyword(s):  
Light Sheet ◽  
Pulse Frequency ◽  
Light Sheet Microscopy

scholarly journals Spatiotemporal co-dependency between macrophages and exhausted CD8+ T cells in cancer

2021 ◽  
Author(s):  
Kelly Kersten ◽  
Kenneth H Hu ◽  
Alexis J Combes ◽  
Bushra Samad ◽  
Tory Harwin ◽  
...  
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Light Sheet ◽  
Light Sheet Microscopy ◽  
Spatially Resolved ◽  
Hypoxic Conditions ◽  
Synaptic Interactions ◽  
Positive Feedback Circuit ◽  
Major Impediment

T cell exhaustion is a major impediment to anti-tumor immunity. However, it remains elusive how other immune cells in the tumor microenvironment (TME) contribute to this dysfunctional state. Here we show that the biology of tumor-associated macrophages (TAM) and exhausted T cells (Tex) in the TME is extensively linked. We demonstrate that in vivo depletion of TAM reduces exhaustion programs in tumor-infiltrating CD8+ T cells and reinvigorates their effector potential. Reciprocally, transcriptional and epigenetic profiling reveals that Tex express factors that actively recruit monocytes to the TME and shape their differentiation. Using lattice light sheet microscopy, we show that TAM and CD8+ T cells engage in unique long-lasting antigen-specific synaptic interactions that fail to activate T cells, but prime them for exhaustion, which is then accelerated in hypoxic conditions. Spatially resolved sequencing supports a spatiotemporal self-enforcing positive feedback circuit that is aligned to protect rather than destroy a tumor.

scholarly journals Coupling optogenetics and light-sheet microscopy to study signal transduction in vivo

2017 ◽  
Vol 145 ◽  
pp. S70-S71
Author(s):  
Prameet Kaur ◽  
Timothy E. Saunders ◽  
Nicholas Tolwinski
Keyword(s):  
Signal Transduction ◽  
Light Sheet ◽  
Light Sheet Microscopy

Functional in vivo imaging using fluorescence lifetime light sheet microscopy (Conference Presentation)

2019 ◽  
Author(s):  
Simon M. Ameer-Beg ◽  
Claire A. Mitchell ◽  
Simon P. Poland ◽  
Robert D. Knight ◽  
Guoqing Wang ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
In Vivo Imaging ◽  
Light Sheet ◽  
Light Sheet Microscopy

scholarly journals Brain-wide genetic mapping identifies the indusium griseum as a prenatal target of pharmacologically unrelated psychostimulants

2019 ◽  
Vol 116 (51) ◽  
pp. 25958-25967
Author(s):  
Janos Fuzik ◽  
Sabah Rehman ◽  
Fatima Girach ◽  
Andras G. Miklosi ◽  
Solomiia Korchynska ◽  
...  
Keyword(s):  
Granule Cells ◽  
Light Sheet ◽  
Pregnant Mouse ◽  
Tissue Imaging ◽  
Drug Induced ◽  
Genetic Ablation ◽  
Glutamatergic Neurons ◽  
Light Sheet Microscopy ◽  
Adult Mice ◽  
Dramatic Rise

Psychostimulant use is an ever-increasing socioeconomic burden, including a dramatic rise during pregnancy. Nevertheless, brain-wide effects of psychostimulant exposure are incompletely understood. Here, we performed Fos-CreERT2–based activity mapping, correlated for pregnant mouse dams and their fetuses with amphetamine, nicotine, and caffeine applied acutely during midgestation. While light-sheet microscopy-assisted intact tissue imaging revealed drug- and age-specific neuronal activation, the indusium griseum (IG) appeared indiscriminately affected. By using GAD67gfp/+mice we subdivided the IG into a dorsolateral domain populated by γ-aminobutyric acidergic interneurons and a ventromedial segment containing glutamatergic neurons, many showing drug-induced activation and sequentially expressing Pou3f3/Brn1 and secretagogin (Scgn) during differentiation. We then combined Patch-seq and circuit mapping to show that the ventromedial IG is a quasi-continuum of glutamatergic neurons (IG-Vglut1+) reminiscent of dentate granule cells in both rodents and humans, whose dendrites emanate perpendicularly toward while their axons course parallel with the superior longitudinal fissure. IG-Vglut1+neurons receive VGLUT1+and VGLUT2+excitatory afferents that topologically segregate along their somatodendritic axis. In turn, their efferents terminate in the olfactory bulb, thus being integral to a multisynaptic circuit that could feed information antiparallel to the olfactory–cortical pathway. In IG-Vglut1+neurons, prenatal psychostimulant exposure delayed the onset of Scgn expression. Genetic ablation ofScgnwas then found to sensitize adult mice toward methamphetamine-induced epilepsy. Overall, our study identifies brain-wide targets of the most common psychostimulants, among whichScgn+/Vglut1+neurons of the IG link limbic and olfactory circuits.

In vivo light‐sheet microscopy resolves localisation patterns of FSD1 , a superoxide dismutase with function in root development and osmoprotection

2020 ◽  
Vol 44 (1) ◽  
pp. 68-87 ◽  
Author(s):  
Petr Dvořák ◽  
Yuliya Krasylenko ◽  
Miroslav Ovečka ◽  
Jasim Basheer ◽  
Veronika Zapletalová ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Root Development ◽  
Light Sheet ◽  
Light Sheet Microscopy
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