The University of Texas at Dallas, Department of Bioengineering, Richardson, Texas, United States.
The University of Texas at Dallas, Center for Imaging and Surgical Innovation, Richardson, Texas, United States.
J Biomed Opt. 2024 Jan;29(1):016005. doi: 10.1117/1.JBO.29.1.016005. Epub 2024 Jan 18.
Polarized hyperspectral microscopes with the capability of full Stokes vector imaging have potential for many biological and medical applications.
The aim of this study is to investigate polarized hyperspectral imaging (PHSI) for improving the visualization of collagen fibers, which is an important biomarker related to tumor development, and improving the differentiation of normal and tumor cells on pathologic slides.
We customized a polarized hyperspectral microscopic imaging system comprising an upright microscope with a motorized stage, two linear polarizers, two liquid crystal variable retarders (LCVRs), and a compact SnapScan hyperspectral camera. The polarizers and LCVRs worked in tandem with the hyperspectral camera to acquire polarized hyperspectral images, which were further used to calculate four Stokes vectors: , , , and . Synthetic RGB images of the Stokes vectors were generated for the visualization of cellular components in PHSI images. Regions of interest of collagen, normal cells, and tumor cells in the synthetic RGB images were selected, and spectral signatures of the selected components were extracted for comparison. Specifically, we qualitatively and quantitatively investigated the enhanced visualization and spectral characteristics of dense fibers and sparse fibers in normal stroma tissue, fibers accumulated within tumors, and fibers accumulated around tumors.
By employing our customized polarized hyperspectral microscope, we extract the spectral signatures of Stokes vector parameters of collagen as well as of tumor and normal cells. The measurement of Stokes vector parameters increased the image contrast of collagen fibers and cells in the slides.
With the spatial and spectral information from the Stokes vector data cubes (, , , and ), our PHSI microscope system enhances the visualization of tumor cells and tumor microenvironment components, thus being beneficial for pathology and oncology.
具有全斯托克斯向量成像功能的偏振高光谱显微镜在许多生物和医学应用中具有潜力。
本研究旨在研究偏振高光谱成像（PHSI）在改善与肿瘤发展相关的重要生物标志物胶原纤维可视化以及改善病理切片上正常和肿瘤细胞的区分方面的应用。
我们定制了一个偏振高光谱显微镜成像系统，该系统包括一个带电动载物台的直立显微镜、两个线偏振器、两个液晶可变延迟器（LCVR）和一个紧凑的 SnapScan 高光谱相机。偏振器和 LCVR 与高光谱相机协同工作，以获取偏振高光谱图像，进一步用于计算四个斯托克斯向量： 、 、 、 。合成的 Stokes 向量 RGB 图像用于 PHSI 图像中细胞成分的可视化。在合成的 RGB 图像中选择胶原、正常细胞和肿瘤细胞的感兴趣区域，并提取所选成分的光谱特征进行比较。具体来说，我们定性和定量地研究了正常基质组织中密集纤维和稀疏纤维、肿瘤内积聚的纤维以及肿瘤周围积聚的纤维的增强可视化和光谱特征。
通过使用我们定制的偏振高光谱显微镜，我们提取了胶原以及肿瘤和正常细胞的 Stokes 向量参数的光谱特征。斯托克斯向量参数的测量增加了载玻片上胶原纤维和细胞的图像对比度。
通过来自 Stokes 向量数据立方体（ 、 、 、 ）的空间和光谱信息，我们的 PHSI 显微镜系统增强了肿瘤细胞和肿瘤微环境成分的可视化，从而有利于病理学和肿瘤学。
