Millisecond lifetime imaging with a europium complex using a commercial confocal microscope under one or two-photon excitation

2014 ◽  
Vol 5 (9) ◽  
pp. 3475-3485 ◽  
Author(s):  
Alexei Grichine ◽  
Alexandre Haefele ◽  
Simon Pascal ◽  
Alain Duperray ◽  
Richard Michel ◽  
...  
Keyword(s):  
Confocal Microscope ◽  
Europium Complex ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Photon Excitation ◽  
Long Lifetime ◽  
Two Photon Excitation

We demonstrate that a commercial confocal microscope can perform time-gated and long lifetime imaging in the μs to ms range under classical one photon or nonlinear two photon excitation.

Applications of Two-Photon Excitation Fluorescence Lifetime Imaging in Tumor Diagnosis

2018 ◽  
Vol 45 (2) ◽  
pp. 0207010
Author(s):  
李慧 Li Hui ◽  
夏先园 Xia Xianyuan ◽  
陈廷爱 Chen Tingai ◽  
余佳 Yu Jia ◽  
李曦 Li Xi ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Tumor Diagnosis ◽  
Fluorescence Lifetime Imaging ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Photon Excitation ◽  
Two Photon Excitation

scholarly journals Depth Penetration and Detection of pH Gradients in Biofilms by Two-Photon Excitation Microscopy

1999 ◽  
Vol 65 (8) ◽  
pp. 3502-3511 ◽  
Author(s):  
Jurrien M. Vroom ◽  
Kees J. De Grauw ◽  
Hans C. Gerritsen ◽  
David J. Bradshaw ◽  
Philip D. Marsh ◽  
...  
Keyword(s):  
Mixed Culture ◽  
Fluorescence Lifetime ◽  
Laser Scanning Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Ph Gradients ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Microbial Biofilms ◽  
Photon Excitation ◽  
Two Photon Excitation

ABSTRACT Deep microbial biofilms are a major problem in many industrial, environmental, and medical settings. Novel approaches are needed to understand the structure and metabolism of these biofilms. Two-photon excitation microscopy (TPE) and conventional confocal laser scanning microscopy (CLSM) were compared quantitatively for the ability to visualize bacteria within deep in vitro biofilms. pH gradients within these biofilms were determined by fluorescence lifetime imaging, together with TPE. A constant-depth film fermentor (CDFF) was inoculated for 8 h at 50 ml · h−1 with a defined mixed culture of 10 species of bacteria grown in continuous culture. Biofilms of fixed depths were developed in the CDFF for 10 or 11 days. The microbial compositions of the biofilms were determined by using viable counts on selective and nonselective agar media; diverse mixed-culture biofilms developed, including aerobic, facultative, and anaerobic species. TPE was able to record images four times deeper than CLSM. Importantly, in contrast to CLSM images, TPE images recorded deep within the biofilm showed no loss of contrast. The pH within the biofilms was measured directly by means of fluorescence lifetime imaging; the fluorescence decay of carboxyfluorescein was correlated with biofilm pH and was used to construct a calibration curve. pH gradients were detectable, in both the lateral and axial directions, in steady-state biofilms. When biofilms were overlaid with 14 mM sucrose for 1 h, distinct pH gradients developed. Microcolonies with pH values of below pH 3.0 were visible, in some cases adjacent to areas with a much higher pH (>5.0). TPE allowed resolution of images at significantly greater depths (as deep as 140 μm) than were possible with CLSM. Fluorescence lifetime imaging allowed the in situ, real-time imaging of pH and the detection of sharp gradients of pH within microbial biofilms.

scholarly journals Two-photon excitation with pico-second fluorescence lifetime imaging to detect nuclear association of flavanols

2012 ◽  
Vol 719 ◽  
pp. 68-75 ◽  
Author(s):  
Irene Mueller-Harvey ◽  
Walter Feucht ◽  
Juergen Polster ◽  
Lucie Trnková ◽  
Pierre Burgos ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Photon Excitation ◽  
Nuclear Association ◽  
Two Photon Excitation
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