scholarly journals 3D-resolved fluorescence and phosphorescence lifetime imaging using temporal focusing wide-field two-photon excitation

2012 ◽  
Vol 20 (24) ◽  
pp. 26219 ◽  
Author(s):  
Heejin Choi ◽  
Dimitrios S. Tzeranis ◽  
Jae Won Cha ◽  
Philippe Clémenceau ◽  
Sander J. G. de Jong ◽  
...  
Keyword(s):  
Wide Field ◽  
Phosphorescence Lifetime ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Photon Excitation ◽  
Temporal Focusing ◽  
Phosphorescence Lifetime Imaging ◽  
Fluorescence And Phosphorescence ◽  
Two Photon Excitation

scholarly journals Two-photon patterned photostimulation with low-power, high-efficiency and reliable single-cell optogenetic control

2022 ◽  
Author(s):  
Yifan Wang ◽  
Yao Zheng ◽  
Yongxian Xu ◽  
Rongrong Li ◽  
Yameng Zheng ◽  
...  
Keyword(s):  
Low Power ◽  
Single Cell ◽  
Laser Power ◽  
High Efficiency ◽  
Excitation Density ◽  
Two Photon ◽  
Photon Excitation ◽  
Temporal Focusing ◽  
Patterned Illumination ◽  
Two Photon Excitation

Two-photon optogenetics enables selectively stimulating individual cells for manipulating neuronal ensembles. As the general photostimulation strategy, the patterned two-photon excitation has enabled millisecond-timescale activation for single or multiple neurons, but its activation efficiency is suffered from high laser power due to low beam-modulation efficiency. Here, we develop a high-efficiency beam-shaping method based on the Gerchberg-Saxton (GS) algorithm with spherical-distribution initial phase (GSSIP) to reduce the patterned two-photon excitation speckles and intensity. It can well control the phase of shaped beams to attain speckle-free accurate patterned illumination with an improvement of 44.21% in the modulation efficiency compared with that of the traditional GS algorithm. A combination of temporal focusing and the GSSIP algorithm (TF-GSSIP) achieves patterned focusing through 500-μm-thickness mouse brain slices, which is 2.5 times deeper than the penetration depth of TF-GS with the same signal-to-noise ratio (SNR). With our method, the laser power can be reduced to only 55.56% of that with traditional method (the temporal focusing with GS, TF-GS) to reliably evoke GCaMP6s response in C1V1-expressing cultured neurons with single-cell resolution. Besides, the photostimulation efficiency is remarkably increased by 80.19% at the same excitation density of 0.27 mW/μm2. This two-photon stimulation method with low-power, reliable and patterned illumination may pave the way for analyzing neural circuits and neural coding and decoding mechanism.

scholarly journals Monitoring mitochondrial viscosity with anticancer phosphorescent Ir(iii) complexes via two-photon lifetime imaging

2019 ◽  
Vol 10 (5) ◽  
pp. 1285-1293 ◽  
Author(s):  
Liang Hao ◽  
Zhi-Wei Li ◽  
Dong-Yang Zhang ◽  
Liang He ◽  
Wenting Liu ◽  
...  
Keyword(s):  
Mitochondrial Dysfunction ◽  
Phosphorescence Lifetime ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Phosphorescence Lifetime Imaging

A theranostic Ir(iii) complex is developed to induce mitochondrial dysfunction and monitor mitochondrial viscosity quantitatively via two-photon phosphorescence lifetime imaging.

scholarly journals Scanless two-photon excitation with temporal focusing

2020 ◽  
Vol 17 (6) ◽  
pp. 571-581 ◽  
Author(s):  
Eirini Papagiakoumou ◽  
Emiliano Ronzitti ◽  
Valentina Emiliani
Keyword(s):  
Two Photon ◽  
Photon Excitation ◽  
Temporal Focusing ◽  
Two Photon Excitation

scholarly journals Two-photon phosphorescence lifetime imaging of cells and tissues using a long-lived cyclometallated Npyridyl^Cphenyl^Npyridyl Pt(ii) complex

RSC Advances ◽  
2014 ◽  
Vol 4 (66) ◽  
pp. 35003-35008 ◽  
Author(s):  
Elizabeth Baggaley ◽  
Igor V. Sazanovich ◽  
J. A. Gareth Williams ◽  
John W. Haycock ◽  
Stanley W. Botchway ◽  
...  
Keyword(s):  
Time Frame ◽  
Phosphorescence Lifetime ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Bio Imaging ◽  
Phosphorescence Lifetime Imaging

The ‘longer’ picture: emission bio-imaging over microsecond time frame with scanning, multi-photon posphorescence-lifetime-imaging-microscopy (PLIM).

scholarly journals Wide field light-sheet microscopy with lens-axicon controlled two-photon Bessel beam illumination

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Sota Takanezawa ◽  
Takashi Saitou ◽  
Takeshi Imamura
Keyword(s):  
High Speed ◽  
Bessel Beam ◽  
Light Sheet ◽  
Time Lapse ◽  
Whole Body ◽  
Wide Field ◽  
Two Photon ◽  
Light Sheet Microscopy ◽  
Photon Excitation ◽  
Two Photon Excitation

AbstractTwo-photon excitation can lower phototoxicity and improve penetration depth, but its narrow excitation range restricts its applications in light-sheet microscopy. Here, we propose simple illumination optics, a lens-axicon triplet composed of an axicon and two convex lenses, to generate longer extent Bessel beams. This unit can stretch the beam full width at half maximum of 600–1000 μm with less than a 4-μm waist when using a 10× illumination lens. A two-photon excitation digital scanned light-sheet microscope possessing this range of field of view and ~2–3-μm axial resolution is constructed and used to analyze the cellular dynamics over the whole body of medaka fish. We demonstrate long-term time-lapse observations over several days and high-speed recording with ~3 mm3 volume per 4 s of the embryos. Our system is minimal and suppresses laser power loss, which can broaden applications of two-photon excitation in light-sheet microscopy.

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