Other Journal Details
Mandatory Fields
Ryder, A.G; Power, S; Glynn, T.J; Morrison, J.J.;
Proc SPIE - Int. Soc. Opt. Eng.
Time-domain measurement of fluorescence lifetime variation with pH
Optional Fields
Fluorescence Time-resolved Lifetime pH
Advances in the design and miniaturization of the lasers and electronics required for Time Correlated Single Photon Counting (TCSPC) measurement of fluorescence lifetime have simplified the use of the time domain method. We have assembled a compact portable system that is capable of measuring lifetimes down to ~200 ps (with deconvolution) and that can operate at a range of excitation and emission wavelengths. The excitation sources are pulsed LEDs and laser diodes with a maximum pulse rate of 40 MHz and are easily interchanged. Furthermore, the development of violet and blue GaN LEDs and laser diodes is expanding the range of fluorophores available for fluorescence lifetime measurement of ion concentrations. pH sensitive fluorophores have a wide range of biological and clinical applications. The use of fluorescence lifetime rather than intensity to measure pH has a number of advantages including the reduction of effects due to photobleaching, scattering, and intensity variations in the excitation source. Using our compact TCSPC instrumentation we have measured the dependence of fluorescence lifetimes on pH for a range of dyes in phosphate buffer over the physiologically important range of 6.0 to 8.0. Most dyes exhibit only a small variation in lifetime (<1.0 ns) over the 6.0 to 8.0 pH range; however, acridine exhibits a large variation in lifetime and hence shows promise as a pH indicator.
Grant Details
The work was funded by grants from the Irish Higher Education Authority, under its Programme for Research in Third Level Institutions, and by a Millennium Research Fund grant from NUI-Galway.
Publication Themes
Biomedical Science and Engineering, Informatics, Physical and Computational Sciences