scholarly journals A lifetime in photochemistry; some ultrafast measurements on singlet states

Author(s):  
David Phillips

We describe here the development of time-correlated single-photon counting techniques from the early use of spark discharge lamps as light sources through to the use of femtosecond mode-locked lasers through the personal work of the author. We used laser-excited fluorescence in studies on energy migration and rotational relaxation in synthetic polymer solutions, in biological probe molecules and in supersonic jet expansions. Time-correlated single-photon counting was the first method used in early fluorescence lifetime imaging microscopy (FLIM), and we outline the development of this powerful technique, with a comparison of techniques including wide-field microscopy. We employed these modern forms of FLIM to study single biological cells, and applied FLIM also to gain an understanding the distribution in tissue, and fates of photosensitizer molecules used in photodynamic therapy. We also describe the uses and instrumental design of laser systems for the study of ultrafast time-resolved vibrational spectroscopy.

2013 ◽  
Vol 368 (1611) ◽  
pp. 20120035 ◽  
Author(s):  
X. Michalet ◽  
R. A. Colyer ◽  
G. Scalia ◽  
A. Ingargiola ◽  
R. Lin ◽  
...  

Two optical configurations are commonly used in single-molecule fluorescence microscopy: point-like excitation and detection to study freely diffusing molecules, and wide field illumination and detection to study surface immobilized or slowly diffusing molecules. Both approaches have common features, but also differ in significant aspects. In particular, they use different detectors, which share some requirements but also have major technical differences. Currently, two types of detectors best fulfil the needs of each approach: single-photon-counting avalanche diodes (SPADs) for point-like detection, and electron-multiplying charge-coupled devices (EMCCDs) for wide field detection. However, there is room for improvements in both cases. The first configuration suffers from low throughput owing to the analysis of data from a single location. The second, on the other hand, is limited to relatively low frame rates and loses the benefit of single-photon-counting approaches. During the past few years, new developments in point-like and wide field detectors have started addressing some of these issues. Here, we describe our recent progresses towards increasing the throughput of single-molecule fluorescence spectroscopy in solution using parallel arrays of SPADs. We also discuss our development of large area photon-counting cameras achieving subnanosecond resolution for fluorescence lifetime imaging applications at the single-molecule level.


Author(s):  
Mike Bruce ◽  
Rama R. Goruganthu ◽  
Shawn McBride ◽  
David Bethke ◽  
J.M. Chin

Abstract For time resolved hot carrier emission from the backside, an alternate approach is demonstrated termed single point PICA. The single point approach records time resolved emission from an individual transistor using time-correlated-single-photon counting and an avalanche photo-diode. The avalanche photo-diode has a much higher quantum efficiency than micro-channel plate photo-multiplier tube based imaging cameras typically used in earlier approaches. The basic system is described and demonstrated from the backside on a ring oscillator circuit.


Author(s):  
Maria Concetta Maccarone ◽  
Giovanni La Rosa ◽  
Osvaldo Catalano ◽  
Salvo Giarrusso ◽  
Alberto Segreto ◽  
...  

AbstractUVscope is an instrument, based on a multi-pixel photon detector, developed to support experimental activities for high-energy astrophysics and cosmic ray research. The instrument, working in single photon counting mode, is designed to directly measure light flux in the wavelengths range 300-650 nm. The instrument can be used in a wide field of applications where the knowledge of the nocturnal environmental luminosity is required. Currently, one UVscope instrument is allocated onto the external structure of the ASTRI-Horn Cherenkov telescope devoted to the gamma-ray astronomy at very high energies. Being co-aligned with the ASTRI-Horn camera axis, UVscope can measure the diffuse emission of the night sky background simultaneously with the ASTRI-Horn camera, without any interference with the main telescope data taking procedures. UVscope is properly calibrated and it is used as an independent reference instrument for test and diagnostic of the novel ASTRI-Horn telescope.


2021 ◽  
Author(s):  
Julia R. Lazzari-Dean ◽  
Evan W. Miller

AbstractBackgroundMembrane potential (Vmem) exerts physiological influence across a wide range of time and space scales. To study Vmem in these diverse contexts, it is essential to accurately record absolute values of Vmem, rather than solely relative measurements.Materials & MethodsWe use fluorescence lifetime imaging of a small molecule voltage sensitive dye (VF2.1.Cl) to estimate mV values of absolute membrane potential.ResultsWe test the consistency of VF2.1.Cl lifetime measurements performed on different single photon counting instruments and find that they are in striking agreement (differences of <0.5 ps/mV in the slope and <50 ps in the y-intercept). We also demonstrate that VF2.1.Cl lifetime reports absolute Vmem under two-photon (2P) illumination with better than 20 mV of Vmem resolution, a nearly 10-fold improvement over other lifetime-based methods.ConclusionsWe demonstrate that VF-FLIM is a robust and portable metric for Vmem across imaging platforms and under both one-photon and two-photon illumination. This work is a critical foundation for application of VF-FLIM to record absolute membrane potential signals in thick tissue.


2016 ◽  
Vol 16 (10) ◽  
pp. 3827-3833 ◽  
Author(s):  
Davide Tamborini ◽  
Mauro Buttafava ◽  
Alessandro Ruggeri ◽  
Franco Zappa

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