The main objective of this research project is to implement a methodology that allows the modulation of physiological cell responses at the single cell level by means of a photodynamic approach. The photodynamic effect produces Reactive Oxygen Species (ROS) when a photosensitizing compound is optically excited in the presence of molecular oxygen. ROS at low concentrations are produced by the cells themselves as physiological signaling agents, so a low-dose photodynamic treatment activates the same cell signaling pathways that rely on endogenous ROS generation. Making use of this photodynamic methodology it is anticipated that a whole range of cell responses (e.g. proliferation, differentiation, reprogramming, etc.) will be elicited on in vitro studies depending on the photodynamic dose. The specific objectives of this proposal will be the study of the multiple cell responses induced by the photodynamic treatment.
This project goals will be achieved by making use of a femtosecond pulsed laser microscopic system that allows the photodynamic treatment of a single cell at a time. This laser setup provides with submicron spatial accuracy and very fine tuning of the delivered light dose. Different assessment methodologies, ranging from bright field imaging to fluorescence microscopy and biochemical analysis to cite a few, will be employed to study cell behaviour during and after the treatments.
The obtained results will be most relevant because previous studies on physiological responses to photodynamic ROS exposure are very scarce and none has been done at the single cell level. Also a reliable methodology to induce cell proliferation or modulation on a desired cell will be of utmost importance in the Biomedical field.