Development of the fast astigmatic auto-focus microscope system

In this paper, a fast auto-focus microscope system was developed based on the astigmatic method. A collimated infrared laser beam was employed in the infinite-corrected microscope optical axis by the beam splitter and reflected by the sample surface. By embedding an astigmatic lens in the system, the reflected laser beam has different focal lengths in the sagittal and tangential planes. As the microscope's relative distance varies, the reflected laser beam shape also varies and can be detected by an embedded four-quadrant photodiode, i.e., the focus error signal (FES) can be found. Then, a fast auto-focus system can be realized by converting the FES to the microscope's defocus distance. We designed an astigmatic auto-focus system for a 20× objective lens with a ±50 µm working range, and this system could also be used for 10× and 5× objectives with ±200 µm and ±800 µm working ranges, respectively.