Biophotonics Laboratory, Immunology Frontier Research Center (IFReC), Osaka University, Yamadaoka 3-1, Suita, Osaka, 565-0871, Japan.
Open and Transdisciplinary Research Institute (OTRI), Osaka University, Yamadaoka 3-1, Suita, Osaka, 565-0871, Japan.
Sci Rep. 2023 Feb 22;13(1):3129. doi: 10.1038/s41598-023-29259-8.
The monitoring of dynamic cellular behaviors remains a technical challenge for most established techniques used nowadays for single-cell analysis, as most of them are either destructive, or rely on labels that can affect the long-term functions of cells. We employ here label-free optical techniques to non-invasively monitor the changes that occur in murine naive T cells upon activation and subsequent differentiation into effector cells. Based on spontaneous Raman single-cell spectra, we develop statistical models that allow the detection of activation, and employ non-linear projection methods to delineate the changes occurring over a several day period spanning early differentiation. We show that these label-free results have very high correlation with known surface markers of activation and differentiation, while also providing spectral models that allow the identification of the underlying molecular species that are representative of the biological process under study.
目前用于单细胞分析的大多数技术都存在技术挑战，无法实时监测动态细胞行为，因为它们大多数要么具有破坏性，要么依赖于可能影响细胞长期功能的标记物。在这里，我们采用无标记的光学技术来非侵入性地监测在激活和随后分化为效应细胞过程中，鼠源初始 T 细胞发生的变化。基于自发拉曼单细胞光谱，我们开发了统计模型，以检测细胞的激活，采用非线性投影方法来描绘早期分化过程中几天内发生的变化。我们表明，这些无标记的结果与已知的激活和分化表面标记物具有非常高的相关性，同时还提供了光谱模型，可以识别代表研究中生物过程的潜在分子种类。
