Li Chen, Rai Mani Ratnam, Cai Yuheng, Ghashghaei H Troy, Greenbaum Alon
Joint Department of Biomedical Engineering, North Carolina State University and University of North Carolina at Chapel Hill, Raleigh, NC 27695, USA.
Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27695, USA.
bioRxiv. 2023 Dec 1:2023.11.29.569329. doi: 10.1101/2023.11.29.569329.
Light sheet fluorescence microscopy (LSFM) provides the benefit of optical sectioning coupled with rapid acquisition times for imaging of tissue-cleared specimen. This allows for high-resolution 3D imaging of large tissue volumes. Inherently to LSFM, the quality of the imaging heavily relies on the characteristics of the illumination beam, with the notion that the illumination beam only illuminates a thin section that is being imaged. Therefore, substantial efforts are dedicated to identifying slender, non-diffracting beam profiles that can yield uniform and high-contrast images. An ongoing debate concerns the employment of the most optimal illumination beam; Gaussian, Bessel, Airy patterns and/or others. Comparisons among different beam profiles is challenging as their optimization objective is often different. Given that our large imaging datasets (~0.5TB images per sample) is already analyzed using deep learning models, we envisioned a different approach to this problem by hypothesizing that we can tailor the illumination beam to boost the deep learning models performance. We achieve this by integrating the physical LSFM illumination model after passing through a variable phase mask into the training of a cell detection network. Here we report that the joint optimization continuously updates the phase mask, improving the image quality for better cell detection. Our method's efficacy is demonstrated through both simulations and experiments, revealing substantial enhancements in imaging quality compared to traditional Gaussian light sheet. We offer valuable insights for designing microscopy systems through a computational approach that exhibits significant potential for advancing optics design that relies on deep learning models for analysis of imaging datasets.
光片荧光显微镜(LSFM)具有光学切片的优势,同时采集速度快,可用于对组织透明标本进行成像。这使得能够对大体积组织进行高分辨率三维成像。对于LSFM而言,成像质量在很大程度上依赖于照明光束的特性,其理念是照明光束仅照亮正在成像的薄切片。因此,人们致力于识别细长的、无衍射光束轮廓,以获得均匀且高对比度的图像。目前存在一个争论,即关于使用最优照明光束;高斯光束、贝塞尔光束、艾里光束图案和/或其他光束。不同光束轮廓之间的比较具有挑战性,因为它们的优化目标往往不同。鉴于我们已经使用深度学习模型分析了大量成像数据集(每个样本约0.5TB图像),我们设想了一种不同的方法来解决这个问题,假设我们可以定制照明光束以提高深度学习模型的性能。我们通过将经过可变相位掩膜后的物理LSFM照明模型集成到细胞检测网络的训练中来实现这一点。在此我们报告,联合优化不断更新相位掩膜,提高图像质量以实现更好的细胞检测。我们的方法的有效性通过模拟和实验得到了证明,与传统高斯光片相比,成像质量有了显著提高。我们通过一种计算方法为显微镜系统设计提供了有价值的见解,这种方法在推进依赖深度学习模型进行成像数据集分析的光学设计方面具有巨大潜力。