Digital Cytometry: Extraction of Forward and Side Scattering Signals From Holotomography.

Jaepil Jo, Herve Hugonnet, Mahn Jae Lee, YongKeun Park
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Abstract

Flow cytometry is a cornerstone technique in medical and biological research, providing crucial information about cell size and granularity through forward scatter (FSC) and side scatter (SSC) signals. Despite its widespread use, the precise relationship between these scatter signals and corresponding microscopic images remains underexplored. Here, we investigate this intrinsic relationship by utilizing scattering theory and holotomography, a three-dimensional quantitative phase imaging (QPI) technique. We demonstrate the extraction of FSC and SSC signals from individual, unlabeled cells by analyzing their three-dimensional refractive index distributions obtained through holotomography. Additionally, we introduce a method for digital windowing of SSC signals to facilitate effective segmentation and morphology-based cell type classification. Our approach bridges the gap between flow cytometry and microscopic imaging, offering a new perspective on analyzing cellular characteristics with high accuracy and without the need for labeling.

数字细胞测量:从 Holotomography 提取正向和侧向散射信号。
流式细胞术是医学和生物学研究的基础技术,通过前向散射(FSC)和侧向散射(SSC)信号提供关于细胞大小和粒度的重要信息。尽管它被广泛使用,但这些散射信号和相应的显微图像之间的确切关系仍未得到充分探索。在这里,我们利用散射理论和全息层析成像,一种三维定量相位成像(QPI)技术来研究这种内在关系。我们展示了FSC和SSC信号的提取,从单独的,未标记的细胞通过分析他们的三维折射率分布获得全息成像。此外,我们还介绍了一种SSC信号的数字窗口方法,以促进有效的分割和基于形态的细胞类型分类。我们的方法弥合了流式细胞术和显微成像之间的差距,为高精度分析细胞特征提供了新的视角,无需标记。
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