Muddiman Ryan, Harkin Sarah, Butler Marion, Hennelly Bryan
Department of Electronic Engineering, Maynooth University, Kildare, Ireland.
Department of Biology, Maynooth University, Kildare, Ireland.
J Biophotonics. 2025 Mar;18(3):e202400382. doi: 10.1002/jbio.202400382. Epub 2025 Jan 18.
Broadband CARS is a coherent Raman scattering technique that provides access to the full biological vibrational spectrum within milliseconds, facilitating the recording of widefield hyperspectral Raman images. In this work, BCARS hyperspectral images of unstained cells from two different cell lines of immune lineage (T cell [Jurkat] and pDCs [CAL-1]) were recorded and analyzed using multivariate statistical algorithms in order to determine the spectral differences between the cells. A classifier was trained which could distinguish the known cells with a 97% out-of-bag accuracy. The classifier was then applied to unlabeled samples containing a mixture of the two cell types on the same coverslip. This work demonstrates single-cell analysis of pDCs (CAL-1) and T cells (Jurkat) using BCARS. This approach enables an initial validation of cellular classification. We further demonstrate the capability of BCARS cell classification using single spectra of 5 ms acquisition time.
宽带相干反斯托克斯拉曼散射（BCARS）是一种相干拉曼散射技术，可在数毫秒内获取完整的生物振动光谱，有助于记录宽场高光谱拉曼图像。在这项工作中，使用多变量统计算法记录并分析了来自两种不同免疫谱系细胞系（T细胞[Jurkat]和浆细胞样树突状细胞[pDCs][CAL-1]）的未染色细胞的BCARS高光谱图像，以确定细胞之间的光谱差异。训练了一个分类器，其可以以97%的袋外准确率区分已知细胞。然后将该分类器应用于同一盖玻片上包含两种细胞类型混合物的未标记样品。这项工作展示了使用BCARS对浆细胞样树突状细胞（CAL-1）和T细胞（Jurkat）进行单细胞分析。这种方法能够对细胞分类进行初步验证。我们进一步展示了使用采集时间为5毫秒的单光谱进行BCARS细胞分类的能力。
