Research on fluorescence image acquisition device and fluorescence image processing technology based on digital PCR chip

In order to solve the problem of inaccurate droplet count and negative and positive identification in image processing by existing digital PCR technology, this paper constructed a fluorescence image acquisition device and fluorescence image processing method based on digital PCR chip, which can process images from the chip. The device includes automatic image acquisition, microdrop counting and positive and negative discrimination functions. The fluorescence excitation module excites uniform excitation light on the chip, and the gray and white camera and photography system are used to capture fluorescence images, while the excitation and acquisition module shows good performance in ZEMAX. Through the collection of three sets of chip images, the experimental results show that the microdrop counting accuracy reaches $99.78 \%$, the positive recognition rate reaches $\mathbf{9 9 . 8 \%}$, and the concentration discrimination error is only $0.13 \%$. The fluorescence acquisition device and the fluorescence treatment method proposed in this paper have a high recognition rate of yin-yang microdroplet detection, reduce the interference of fluorescence noise, further improve the accuracy of the results, and have broad application prospects for nucleic acid quantitative and trace detection.