Intelligent Beam Optimization for Light-Sheet Fluorescence Microscopy Through Deep Learning

Light-sheet fluorescence microscopy (LSFM) provides the benefit of optical sectioning coupled with rapid acquisition times, enabling high-resolution 3-dimensional imaging of large tissue-cleared samples. Inherent to LSFM, the quality of the imaging heavily relies on the characteristics of the illumination beam, which only illuminates a thin section of the sample. Therefore, substantial efforts are dedicated to identifying slender, nondiffracting beam profiles that yield uniform and high-contrast images. An ongoing debate concerns the identification of optimal illumination beams for different samples: Gaussian, Bessel, Airy patterns, and/or others. However, comparisons among different beam profiles are challenging as their optimization objectives are often different. Given that our large imaging datasets (approximately 0.5 TB of images per sample) are already analyzed using deep learning models, we envisioned a different approach to the problem by designing an illumination beam tailored to boost the performance of the deep learning model. We hypothesized that integrating the physical LSFM illumination model (after passing it through a variable phase mask) into the training of a cell detection network would achieve this goal. Here, we report that joint optimization continuously updates the phase mask and results in improved image quality for better cell detection. The efficacy of our method is demonstrated through both simulations and experiments that reveal substantial enhancements in imaging quality compared to the traditional Gaussian light sheet. We discuss how designing microscopy systems through a computational approach provides novel insights for advancing optical design that relies on deep learning models for the analysis of imaging datasets.