Journal of Physical Chemistry A, Vol.119, No.14, 3393-3399, 2015
Singlet Oxygen Phosphorescence Lifetime Imaging Based on a Fluorescence Lifetime Imaging Microscope
The feasibility of singlet oxygen phosphorescence (SOP) lifetime imaging microscope was studied on a modified fluorescence lifetime imaging microscope (FLIM). SOP results from the infrared radiative transition of O-2(a(1)Delta(g) -> X-3 Sigma(-)(g)) and O-2(a(1)Delta(g)) was produced in a C-60 powder sample via photosensitization process. To capture the very weak SOP signal, a dichroic mirror was placed between the objective and tube lens of the FLIM and used to divide the luminescence returning from the sample into two beams: the reflected SOP beam and the transmitted photoluminescence of C-60 (C-60-PL) beam. The C-60-PL beam entered the scanner of the FLIM and followed the normal optical path of the FLIM, while the SOP steered clear, of the scanner and directly entered a finely designed SOP detection channel Confocal C-60-PL images and nonconfocal SOP images were then simultaneously obtained by using laser scanning mode. Experimental results show that (1) under laser scanning mode, the obstacle to confocal SOP imaging is the infrared incompatible scanner, which can be solved by using an infrared compatible scanner. Confocal SOP imaging is also expected to be realized under stage scanning mode when the laser beam is parked and meanwhile a pinhole is added into the SOP detection channel. (2) A great challenge to SOP imaging is its extraordinarily long imaging time, and selecting only a few interesting points from fluorescence images to measure their SOP time-dependent traces may be a correct compromise.