The invention provides a three-dimensional automatic focusing method and device based on multi-
modal data fusion and Kalman filtering and a readable storage medium, and high-precision real-time focusing is realized through multiple steps: collecting sample three-dimensional phase information and different focal plane image data by using a phase and
image sensor, and calculating phase depth and definition; constructing a multi-plane Kalman filtering cluster, modeling the focus state of each focal plane, and predicting the focus by fusing the neighborhood
coupling influence; the focus state is updated and corrected in combination with the image definition and the
phase gradient, and the optimal three-dimensional focus position is determined by means of a collaborative optimization
decision maker; dynamically adjusting the
noise matrix and the
coupling coefficient, and controlling the objective lens to complete focusing. According to the method, through multi-
modal fusion and multi-plane cooperation, the problems of focus drifting and
jumping are effectively solved, the focusing precision and stability of an imaging
system in a complex sample and a dynamic environment are remarkably improved, and the method is suitable for equipment such as a biomedical full-automatic
microscope and a digital
pathological section
scanner.