Method and system for acquisition of fluorescence images of live-cell biological samples

Abstract

A method is disclosed for acquiring a single, in-focus two-dimensional projection image of a live, three-dimensional cell culture sample, with a fluorescence microscope. One or more long-exposure 'Z-sweep' images are obtained, i.e. via a single or series of continuous acquisitions, while moving the Z-focal plane of a camera through the sample, to produce one or more two-dimensional images of fluorescence intensity integrated over the Z-dimension. The acquisition method is much faster than a Z-stack method, which enables higher throughput and reduces the risk of exposing the sample to too much fluorescent light. The long-exposure Z-sweep image(s) is then input into a neural network which has been trained to produce a high-quality (in-focus) two-dimensional projection image of the sample. With these high-quality projection images, biologically relevant analysis metrics can be obtained to describe the fluorescence signal using standard image analysis techniques, such as fluorescence object count and other fluorescence intensity metrics (e.g., mean intensity, texture, etc.).