Active focus stabilization for fluorescence microscopy by tracking actively generated infrared spots

Abstract

Maintaining focus during time-lapse experiments remains an important challenge in optical microscopy. In this paper we present an automated focusing system for fluorescence microscopy that can be used in time-lapse experiments. We provide software for flexibly operating the autofocus system and a complete alignment guide. The system is evaluated using a time-lapse experiment with temperature variations on Tetraspeck beads.