Low-power two-color STED microscopy based on phasor plot analysis

2020 ◽  
Author(s):  
Yue Chen ◽  
Luwei Wang ◽  
Wei Yan ◽  
Junle Qu ◽  
Jun Song
Keyword(s):  
Low Power ◽  
Sted Microscopy ◽  
Plot Analysis ◽  
Phasor Plot

Resolution improvement in STED super-resolution microscopy at low power using a phasor plot approach

Nanoscale ◽  
2018 ◽  
Vol 10 (34) ◽  
pp. 16252-16260 ◽  
Author(s):  
Luwei Wang ◽  
Bingling Chen ◽  
Wei Yan ◽  
Zhigang Yang ◽  
Xiao Peng ◽  
...  
Keyword(s):  
Low Power ◽  
Super Resolution ◽  
Stimulated Emission ◽  
Resolution Limit ◽  
Sted Microscopy ◽  
Microscopy Technique ◽  
Resolution Improvement ◽  
Phasor Plot ◽  
Stimulated Emission Depletion ◽  
Super Resolution Microscopy

Stimulated emission depletion (STED) microscopy is a powerful super-resolution microscopy technique that has achieved significant results in breaking the resolution limit and relevant applications.

scholarly journals Optimization of Advanced Live-Cell Imaging through Red/Near-Infrared Dye Labeling and Fluorescence Lifetime-Based Strategies

2021 ◽  
Vol 22 (20) ◽  
pp. 11092
Author(s):  
Magalie Bénard ◽  
Damien Schapman ◽  
Christophe Chamot ◽  
Fatéméh Dubois ◽  
Guénaëlle Levallet ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Live Cell Imaging ◽  
Near Infrared ◽  
Cell Imaging ◽  
Light Exposure ◽  
Photon Counting ◽  
Live Cell ◽  
Stimulated Emission ◽  
Plot Analysis ◽  
Phasor Plot

Fluorescence microscopy is essential for a detailed understanding of cellular processes; however, live-cell preservation during imaging is a matter of debate. In this study, we proposed a guide to optimize advanced light microscopy approaches by reducing light exposure through fluorescence lifetime (τ) exploitation of red/near-infrared dyes. Firstly, we characterized key instrumental elements which revealed that red/near-infrared laser lines with an 86x (Numerical Aperture (NA) = 1.2, water immersion) objective allowed high transmission of fluorescence signals, low irradiance and super-resolution. As a combination of two technologies, i.e., vacuum tubes (e.g., photomultiplier) and semiconductor microelectronics (e.g., avalanche photodiode), type S, X and R of hybrid detectors (HyD-S, HyD-X and HyD-R) were particularly adapted for red/near-infrared photon counting and τ separation. Secondly, we tested and compared lifetime-based imaging including coarse τ separation for confocal microscopy, fitting and phasor plot analysis for fluorescence lifetime microscopy (FLIM), and lifetimes weighting for enhanced stimulated emission depletion (STED) nanoscopy, in light of red/near-infrared multiplexing. Mainly, we showed that the choice of appropriate imaging approach may depend on fluorochrome number, together with their spectral/lifetime characteristics and STED compatibility. Photon-counting mode and sensitivity of HyDs together with phasor plot analysis of fluorescence lifetimes enabled the flexible and fast imaging of multi-labeled living H28 cells. Therefore, a combination of red/near-infrared dyes labeling with lifetime-based strategies offers new perspectives for live-cell imaging by enhancing sample preservation through acquisition time and light exposure reduction.

scholarly journals Low-Power Two-Color Stimulated Emission Depletion Microscopy for Live Cell Imaging

Biosensors ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 330
Author(s):  
Jia Zhang ◽  
Xinwei Gao ◽  
Luwei Wang ◽  
Yong Guo ◽  
Yinru Zhu ◽  
...  
Keyword(s):  
Low Power ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Stimulated Emission ◽  
Live Cells ◽  
Sted Microscopy ◽  
Resolution Imaging ◽  
Stimulated Emission Depletion

Stimulated emission depletion (STED) microscopy is a typical laser-scanning super-resolution imaging technology, the emergence of which has opened a new research window for studying the dynamic processes of live biological samples on a nanometer scale. According to the characteristics of STED, a high depletion power is required to obtain a high resolution. However, a high laser power can induce severe phototoxicity and photobleaching, which limits the applications for live cell imaging, especially in two-color STED super-resolution imaging. Therefore, we developed a low-power two-color STED super-resolution microscope with a single supercontinuum white-light laser. Using this system, we achieved low-power two-color super-resolution imaging based on digital enhancement technology. Lateral resolutions of 109 and 78 nm were obtained for mitochondria and microtubules in live cells, respectively, with 0.8 mW depletion power. These results highlight the great potential of the novel digitally enhanced two-color STED microscopy for long-term dynamic imaging of live cells.

Phasor plot analysis using low pass filter for high-speed FLIM

2019 ◽  
Author(s):  
Dongeun Kim ◽  
Wonsang Hwang ◽  
Youngjae Won ◽  
Sucbei Moon ◽  
Sang Yoon Lee ◽  
...  
Keyword(s):  
High Speed ◽  
Pass Filter ◽  
Low Pass Filter ◽  
Plot Analysis ◽  
Low Pass ◽  
Phasor Plot

scholarly journals A Method to Quantify FRET Stoichiometry with Phasor Plot Analysis and Acceptor Lifetime Ingrowth

2015 ◽  
Vol 108 (5) ◽  
pp. 999-1002 ◽  
Author(s):  
WeiYue Chen ◽  
Edward Avezov ◽  
Simon C. Schlachter ◽  
Fabrice Gielen ◽  
Romain F. Laine ◽  
...  
Keyword(s):  
Plot Analysis ◽  
Phasor Plot

Less Power, Greater Conflict

2018 ◽  
Vol 49 (1) ◽  
pp. 47-62 ◽  
Author(s):  
Petra C. Schmid
Keyword(s):  
Low Power ◽  
High Power ◽  
Goal Pursuit ◽  
Goal Conflict ◽  
Personal Goals ◽  
Multiple Goals ◽  
Objective Performance ◽  
The Future ◽  
The Way

Abstract. Power facilitates goal pursuit, but how does power affect the way people respond to conflict between their multiple goals? Our results showed that higher trait power was associated with reduced experience of conflict in scenarios describing multiple goals (Study 1) and between personal goals (Study 2). Moreover, manipulated low power increased individuals’ experience of goal conflict relative to high power and a control condition (Studies 3 and 4), with the consequence that they planned to invest less into the pursuit of their goals in the future. With its focus on multiple goals and individuals’ experiences during goal pursuit rather than objective performance, the present research uses new angles to examine power effects on goal pursuit.

An interdependence account of sexism and power: Men’s hostile sexism, biased perceptions of low power, and relationship aggression.

2019 ◽  
Vol 117 (2) ◽  
pp. 338-363 ◽  
Author(s):  
Emily J. Cross ◽  
Nickola C. Overall ◽  
Rachel S. T. Low ◽  
James K. McNulty
Keyword(s):  
Low Power ◽  
Hostile Sexism ◽  
Relationship Aggression
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