A method for microscopic image acquisition based on a sequential slice. The method includes; acquiring a sample of the sequential slice and a navigation image thereof; identifying and labeling the sample of the sequential slice in the navigation image by utilizing methods of image processing and machine learning; placing the sample of the sequential slice in a microscope, establishing a coordinate transformation matrix for a navigation image-microscope actual sampling space coordinate, and navigating and locating a random pixel point in the navigation image to a center of the microscope's visual field; locating the sample of the sequential slice under a low resolution visual field, binding a sample acquisition parameter; based on the binding of the sample acquisition parameter, recording a relationship of relative of locations between a center point of a high resolution acquisition region and a center point after being matched with a sample template.