scholarly journals Multiplexed bioluminescence microscopy via phasor analysis

2021 ◽  
Author(s):  
Zi Yao ◽  
Caroline K. Brennan ◽  
Lorenzo Scipioni ◽  
Hongtao Chen ◽  
Kevin Ng ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Bioluminescence Imaging ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Time Lapse ◽  
Detection Methods ◽  
Transfer Processes ◽  
Time Lapse Imaging ◽  
Luciferase Reporters ◽  
Phasor Analysis

Microscopic bioluminescence imaging has been historically challenging due to a lack of detection methods and easily resolved probes. Here we combine bioluminescence with phasor analysis, an optical method commonly used to distinguish spectrally similar fluorophores. Bioluminescent phasor enabled rapid differentiation of multiple luciferase reporters and resonance energy transfer processes. The merger of bioluminescence and phasor analysis provides a platform for routine, time-lapse imaging of collections of cellular features.

scholarly journals Live imaging of apoptosis in a novel transgenic mouse highlights its role in neural tube closure

2011 ◽  
Vol 195 (6) ◽  
pp. 1047-1060 ◽  
Author(s):  
Yoshifumi Yamaguchi ◽  
Naomi Shinotsuka ◽  
Keiko Nonomura ◽  
Kiwamu Takemoto ◽  
Keisuke Kuida ◽  
...  
Keyword(s):  
Transgenic Mouse ◽  
Neural Tube ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Time Lapse ◽  
Neural Tube Closure ◽  
Caspase Activation ◽  
Fluorescent Reporter ◽  
Time Lapse Imaging ◽  
Tube Closure

Many cells die during development, tissue homeostasis, and disease. Dysregulation of apoptosis leads to cranial neural tube closure (NTC) defects like exencephaly, although the mechanism is unclear. Observing cells undergoing apoptosis in a living context could help elucidate their origin, behavior, and influence on surrounding tissues, but few tools are available for this purpose, especially in mammals. In this paper, we used insulator sequences to generate a transgenic mouse that stably expressed a genetically encoded fluorescence resonance energy transfer (FRET)–based fluorescent reporter for caspase activation and performed simultaneous time-lapse imaging of apoptosis and morphogenesis in living embryos. Live FRET imaging with a fast-scanning confocal microscope revealed that cells containing activated caspases showed typical and nontypical apoptotic behavior in a region-specific manner during NTC. Inhibiting caspase activation perturbed and delayed the smooth progression of cranial NTC, which might increase the risk of exencephaly. Our results suggest that caspase-mediated cell removal facilitates NTC completion within a limited developmental window.

scholarly journals Energy Transfer between Tm-Doped Upconverting Nanoparticles and a Small Organic Dye with Large Stokes Shift

Biosensors ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 9 ◽  
Author(s):  
Anna López de Guereñu ◽  
Philipp Bastian ◽  
Pablo Wessig ◽  
Leonard John ◽  
Michael U. Kumke
Keyword(s):  
Energy Transfer ◽  
Stokes Shift ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Photoluminescence Properties ◽  
Decay Kinetics ◽  
Transfer Processes ◽  
Surface Quenching ◽  
Low Toxicity ◽  
Ion Emission

Lanthanide-doped upconverting nanoparticles (UCNP) are being extensively studied for bioapplications due to their unique photoluminescence properties and low toxicity. Interest in RET applications involving UCNP is also increasing, but due to factors such as large sizes, ion emission distributions within the particles, and complicated energy transfer processes within the UCNP, there are still many questions to be answered. In this study, four types of core and core-shell NaYF4-based UCNP co-doped with Yb3+ and Tm3+ as sensitizer and activator, respectively, were investigated as donors for the Methyl 5-(8-decanoylbenzo[1,2-d:4,5-d′]bis([1,3]dioxole)-4-yl)-5-oxopentanoate (DBD-6) dye. The possibility of resonance energy transfer (RET) between UCNP and the DBD-6 attached to their surface was demonstrated based on the comparison of luminescence intensities, band ratios, and decay kinetics. The architecture of UCNP influenced both the luminescence properties and the energy transfer to the dye: UCNP with an inert shell were the brightest, but their RET efficiency was the lowest (17%). Nanoparticles with Tm3+ only in the shell have revealed the highest RET efficiencies (up to 51%) despite the compromised luminescence due to surface quenching.

Single-excitation dual-color coherent lasing by tuning resonance energy transfer processes in porous structured nanowires

Nanoscale ◽  
2015 ◽  
Vol 7 (37) ◽  
pp. 15091-15098 ◽  
Author(s):  
Zhaona Wang ◽  
Xiaoyu Shi ◽  
Ruomeng Yu ◽  
Sujun Wei ◽  
Qing Chang ◽  
...  
Keyword(s):  
Energy Transfer ◽  
Transfer Process ◽  
Energy Transfer Process ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Random Lasing ◽  
Transfer Processes ◽  
Dual Color

Single-excitation dual-color coherent random lasing was achieved by tuning the resonance energy transfer process in porous structured nanowires.

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