Improving FLIM Resolution with Mean-Shift Super-Resolution Microscopy, an analytical approach

Fluorescence Lifetime Imaging Microscopy (FLIM) is utilized to study the spatial distribution of fluorophores and assess molecular properties within cells or tissues. However, FLIM faces spatial and lifetime blurring due to the convolution of fluorophore light with the microscope’s point spread function, diminishing spatial resolution and affecting lifetime distributions. In this research, we developed a model based on photophysical principles to examine the impact of the Mean-Shift Super-Resolution microscopy (MSSR) approach on FLIM’s spatial and lifetime resolution. Through simulations of isolated fluorophores with lifetimes of 1.0 and 3.0 ns, we demonstrated that MSSR enhances spatial resolution beyond the diffraction limit and reduces lifetime blurring. Experimental validations will be beneficial to further support these findings, potentially contributing to increased accuracy in various medical and biomedical imaging applications.