I will be measuring the fluorescence of DPBF in singlet oxygen detection for its quantification. Can the same procedure be followed for measuring singlet oxygen generated in cell lines?
I do not think, as DPBF is non-soluble in water environment.
DPBF is a non selective probe for singlet oxygen and reactive oxygen species (ROS). While the synthesis of some water-soluble isobenzofuranes can be found, singlet oxygen is usually detected in cells using the commercial Singlet Oxygen Sensor Green® (SOSG) by Molecular Probes. An academid study on the ptohochemical behaviour of Sensor Green both in solution and in mammalian cell by Peter Ogilby from the Center for Oxygen Microscopy and Imaging in Aarhus can be found here: http://onlinelibrary.wiley.com/doi/10.1111/j.1751-1097.2011.00900.x/abstract (Photochemistry and Photobiology, Volume 87, Issue 3, pages 671–679, May/June 2011).
More interesting is that the same Authors have delevoped their own version of this fluorescein-based sensor, the Aarhus Sensor Green (ASG)(http://pubs.acs.org/doi/abs/10.1021/jo500219y, - J. Org. Chem., 2014, 79 (7), pp 3079–3087, DOI: 10.1021/jo500219y).
Ogilby and its coworkers found that ASG readily enters a mammalian cell (i.e., HeLa) and that "the principal advantage of ASG over SOSG is that, at physiological pH values, both ASG and the ASG endoperoxide (ASG-EP) do not themselves photosensitize the production of singlet oxygen. As such, ASG better fits the requirement of being a benign probe."
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