Parker Marina, Mayes Sam A, Browning Craig M, Deal Joshua, Gunn-Mayes Samantha, Rich Thomas C, Leavesley Silas J
Chemical and Biomolecular Engineering, University of South Alabama, AL 36688.
Department of Systems Engineering, University of South Alabama, AL 36688.
Proc SPIE Int Soc Opt Eng. 2022 Jan-Feb;11966. doi: 10.1117/12.2607659. Epub 2022 Mar 2.
Hyperspectral imaging technologies (HSI) have undergone rapid development since their beginning stages. While original applications were in remote sensing, other uses include agriculture, food safety and medicine. HSI has shown great utility in fluorescence microscopy for detecting signatures from many fluorescent molecules; however, acquisitions speeds have been slow due to light losses associated with spectral filtering. Therefore, we designed a novel light emitting diode (LED)-based rapid excitation scanning hyperspectral imaging platform allowing users to obtain simultaneous measurements of fluorescent labels without compromising acquisition speeds. Previously, we reported our results of the optical ray trace simulations and the geometrical capability of designing a multifaceted mirror imaging system as an initial approach to combine light at many wavelengths. The design utilized LEDs and a multifaceted mirror array to combine light sources into a liquid light guide. The computational model was constructed using Monte Carlo optical ray software (TracePro, Lambda Research Corp.). Recent prototype validation results show that when compared to a commercial emission scanning spectral confocal microscope (Zeiss-LSM-980), the novel LED-based excitation scanning HSI prototype successfully detected and separated six fluorescent labels from a custom 6-label African green monkey kidney epithelial cells. We report on the prototype's ability to overcome limitations of acquisition speeds, sensitivity, and specificity present in conventional systems. Future work will evaluate prototype's light losses to determine latent design modifications needed to demonstrate the system's feasibility as a promising solution for overcoming HSI acquisition speeds. This work was supported by NSF award MRI1725937.
高光谱成像技术（HSI）自其起步阶段以来经历了快速发展。虽然其最初的应用领域是遥感，但其他用途还包括农业、食品安全和医学。高光谱成像技术在荧光显微镜中已显示出巨大的实用价值，可用于检测多种荧光分子的特征；然而，由于与光谱滤波相关的光损失，采集速度一直较慢。因此，我们设计了一种基于新型发光二极管（LED）的快速激发扫描高光谱成像平台，使用户能够在不影响采集速度的情况下同时测量荧光标记。此前，我们报告了光线追踪模拟结果以及设计多面镜成像系统作为组合多种波长光的初步方法的几何能力。该设计利用发光二极管和多面镜阵列将光源组合成液体光导。计算模型是使用蒙特卡罗光线软件（TracePro，Lambda Research Corp.）构建的。最近的原型验证结果表明，与商用发射扫描光谱共聚焦显微镜（蔡司 - LSM - 980）相比，基于新型发光二极管的激发扫描高光谱成像原型成功地从定制的6标记非洲绿猴肾上皮细胞中检测并分离出六种荧光标记。我们报告了该原型克服传统系统中存在的采集速度、灵敏度和特异性限制的能力。未来的工作将评估原型的光损失，以确定潜在的设计改进，以证明该系统作为克服高光谱成像采集速度的有前景解决方案的可行性。这项工作得到了美国国家科学基金会（NSF）授予的MRI1725937奖项的支持。
