Optical pickup unit-based fluorescence microscope with autofocus and deep learning capabilities

An autofocus, cost-effective fluorescence microscope instrument is made using a commercially available Blu-ray disc optical pickup head (OPU). This ABFM consists of the optical path, system control and the operation program. The optical path performs both optical imaging and optical autofocus by using a specially designed dichroscope and the focus error signal (FES) technique integrated in the OPU. Because of the autofocusing function, the fluorescent images captured by ABFM with a resolution of 1.0 µm with a 5X objective lens are obtained. Fluorescence images of human corneal epithelial cell-transformed (HCE-T) cells with fluorophore 4,6-diamino-2-phenyl indole (DAPI) are measured by the ABFM, whose result was comparable to those measured by a Nikon ECLIPSE Ti-S fluorescence microscope and a Leica STELLARIS 5 laser scanning confocal microscope (Leica LSCM). A special deep learning model (Deep Fourier Generative Adversarial Network, DFGAN) was trained to enhance the resolution of the HCE-T, improving the resolution by 21 %. Thus, this ABFM can be used as an alternative to commercial fluorescence microscopes for imaging biological samples in the development of cost-effective, high-performance fluorescence microscopy systems.