College of Engineering and Applied Science, University of Wisconsin-Milwaukee, Milwaukee, USA.
College of Health Sciences, University of Wisconsin-Milwaukee, Milwaukee, USA.
Sci Rep. 2022 Feb 15;12(1):2477. doi: 10.1038/s41598-022-06360-y.
Hyperspectral fluorescence imaging is widely used when multiple fluorescent probes with close emission peaks are required. In particular, Fourier transform imaging spectroscopy (FTIS) provides unrivaled spectral resolution; however, the imaging throughput is very low due to the amount of interferogram sampling required. In this work, we apply deep learning to FTIS and show that the interferogram sampling can be drastically reduced by an order of magnitude without noticeable degradation in the image quality. For the demonstration, we use bovine pulmonary artery endothelial cells stained with three fluorescent dyes and 10 types of fluorescent beads with close emission peaks. Further, we show that the deep learning approach is more robust to the translation stage error and environmental vibrations. Thereby, the He-Ne correction, which is typically required for FTIS, can be bypassed, thus reducing the cost, size, and complexity of the FTIS system. Finally, we construct neural network models using Hyperband, an automatic hyperparameter selection algorithm, and compare the performance with our manually-optimized model.
当需要多个发射峰接近的荧光探针时,通常会使用高光谱荧光成像。特别是,傅里叶变换成像光谱(FTIS)提供了无与伦比的光谱分辨率;然而,由于需要对干涉图进行大量采样,因此成像速度非常低。在这项工作中,我们将深度学习应用于 FTIS,并表明可以将干涉图采样减少一个数量级,而图像质量几乎没有明显下降。为此演示,我们使用三种荧光染料和十种发射峰接近的荧光珠对牛肺动脉内皮细胞进行染色。此外,我们还表明,深度学习方法对平移台误差和环境振动更具鲁棒性。因此,可以绕过 FTIS 通常需要的氦氖校正,从而降低 FTIS 系统的成本、尺寸和复杂性。最后,我们使用自动超参数选择算法 Hyperband 构建神经网络模型,并将性能与我们手动优化的模型进行比较。