In cells, hydrogen peroxide (H2O2) is deliberately produced and is key for a healthy cellular metabolism, and an adequate H2O2 production and delivery is required for correct cellular and tissue functions. The majority of this H2O2 production occurs in cellular microdomains, and the signalling actions in these microdomains are restricted to a very confined area. Most of these discoveries were made possible thanks to the use of genetically-encoded fluorescent biosensors that detect the changes in H2O2 concentration, including the HYPER family of biosensors. However, the details of these microdomains are missing because these biosensors rely on conventional fluorescence microscopy methods, which are limited in spatial resolution.
This project aims to overcome the resolution limitation in H2O2 imaging by developing photochromic variants of the HYPER biosensors, known as PCHYPERs, that can be utilized in super-resolution microscopy. This will enable to discern the gradients of metabolic H2O2 at a high resolution, helping identify the role of H2O2 as a cellular messenger in signalling pathways and the functional implications of its disturbance.
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