Method for the analysis of spatial and temporal information of samples by means of optical microscopy

Method for the analysis of spatial and temporal information of samples by means of optical microscopy, comprising the following steps: a) choosing a field of view in a sample; b) scanning the sample in the axial direction by applying an electrical control signal having a first frequency to an electrically tunable liquid lens placed in the detection path, performing a sequential acquisition, at a first acquisition frequency which is higher than the first frequency of the electrical signal, of a stack of images placed on different in-focus planes; c) processing the stack of images for identifying the position of one or more regions of interest in the three-dimensional space under examination of the sample; d) scanning the sample in the axial direction by further applying the electrical signal having a second frequency to the tunable lens, performing a sequential acquisition, at a second acquisition frequency which is lower than the second frequency of the electrical signal, of a temporal series of images with extended depth of field; e) calculating the mean intensity of each region of interest for each image of the temporal series