Fast autofocusing based on pixel difference with the Tanimoto coefficient between images.

Abstract

Focusing objects accurately over short time scales is an essential and nontrivial task for a variety of microscopy applications. In this Letter, an autofocusing algorithm using pixel difference with the Tanimoto coefficient (PDTC) is described to predict the focus. Our method can robustly distinguish differences in clarity among datasets. The generated auto-focusing curves have extremely high sensitivity. A dataset of a defocused stack acquired by an Olympus microscope demonstrates the feasibility of our technique. This work can be applied in full-color microscopic imaging systems and is also valid for single-color imaging.