Rapid lifetime imaging

The time of data collection depends on the complexity of the (5-pulse response. For a single exponential decay phase fluorometry is more rapid. For complex 5-pulse responses, the time of data collection is about the same for the two techniques in pulse fluorometry, a large number of photon events is necessary, and in phase fluorometry, a large number of frequencies has to be selected. It should be emphasized that the short acquisition time for phase shift and modulation ratio measurements at a given frequency is a distinct advantage in several situations, especially for lifetime-imaging spectroscopy. 

Fig. 15 Fluorescence imaging of citrate in a microtiter plate via the EuTc probe by rapid lifetime determination (in false colors). The concentration of EuTc is 50 pmol I. 1 throughout citrate concentrations (from left to right) are 0, 0.16, 0.4, 1.0, 1.6, 4.0, 10.0, 16.0, 20., 40.0, 60.0 and 80.0 pmol L-1...

Fig. 19 Rapid lifetime determination imaging of the activity of glucose oxidase. Gray-scale image of the activity of glucose oxidase (left) and resulting calibration curve (right). Experiments were performed in triplicate (rows). The wells in the images contained, from 1 to 12, glucose oxidase activities of 0 (blank), 135, 54.1, 27.1, 13.5, 5.4, 2.7, 1.35, 0.54, 0.27, 0.14, and 0.05 mU ml. 1 respectively, 100 jiL of a 0.2 mmol L-1 EuTc solution, and 15 jiL of a 277.2 mmol I. 1 glucose solution. The total volume was made up to 200 xL with MOPS buffer...

As distinct from sohd supports such as gold or silver, mercury imparts lateral mobihty to hpid monolayers directly self-assembled on its surface, because of its liquid state. This is demonstrated by rapid spontaneous phase separation, with microdomain formation, in a hpid mixture monolayer self-assembled on top of a DPTL thiolipid monolayer tethered to a mercury microelectrode [30]. The presence of microdomains was directly verified from the images of the distal hpid monolayer obtained using two-photon fluorescence lifetime imaging microscopy. 

Schneider, P. C. and Clegg, R. M. (1997). Rapid acquisition, analysis, and display of fluorescence lifetime-resolved images for real-time applications. Rev. Sci. Instrum. 68, 4107-19. 

Buranachai C, Kamiyama D, Chiba A et al (2008) Rapid frequency-domain FLIM spinning disk confocal microscope lifetime resolution, image improvement and wavelet analysis. JFluoresc 18 929-942... 

Mapping the most frequent rupture forces for different loading rates establishes a so-called dynamic force spectrum of bond strength that images the energy barriers along the force-driven pathwayThese barriers are the determinants of bond/interaction lifetime and strength nnder rapid detachment. 

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