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Minimally invasive measurements of the partial pressure of oxygen (pO2) in biological tissues are of high interest for many applications, including for the monitoring of the irradiation doses applied in radiotherapy and photodynamic therapy (PDT). The latter is an approved treatment of cancers and other conditions that involves a sub-thermal use of light, photosensitizing molecules (photosensitizers “PS”) and oxygen to treat the lesion. Phosphorescence quenching-based methods are of high interest for this monitoring. Unfortunately, virtually all pO2 molecular probes are more or less phototoxic. This feature is not critical when such probes are used for PDT, or in combination with photosensitizers, whereas photodamage is frequently problematic for other applications of such probes. In this chapter, we describe applications of this pO2 monitoring method, which is based on time-resolved measurements of the luminescence emitted by molecular probes that are quenched by oxygen. Furthermore, we pay particular attention to concepts that are in relation with: (i) the oxygen consumption during PDT, (ii) the phototoxicity of oxygen molecular probes and (iii) the photodamages they induce depending on their localization. Finally, strategies to minimize the photodamage induced during such measurements is described in detail.

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