Wang Huijun, Zhang Lu, Huang Jie, Yang Zewen, Fan Chen, Yuan Li, Zhao Hong, Zhang Zhenxi, Liu Xiaolong
State Key Laboratory for Manufacturing System Engineering, Xi'an Jiaotong University, Xi'an 710049, China.
School of Instrument Science and Technology, Xi'an Jiaotong University, Xi'an 710049, China.
Biomed Opt Express. 2024 Mar 15;15(4):2451-2465. doi: 10.1364/BOE.518957. eCollection 2024 Apr 1.
Label-free detection of intracellular substances for living cancer cells remains a significant hurdle in cancer pathogenesis research. Although the sensitivity of light polarization to intracellular substances has been validated, current studies are predominantly focused on tissue lesions, thus label-free detection of substances within individual living cancer cells is still a challenge. The main difficulty is to find specific detection methods along with corresponding characteristic parameters. With refractive index as an endogenous marker of substances, this study proposes a detection method of intracellular refractive index distribution (IRID) for label-free living colon cancer (LoVo) cells. Utilizing the circular depolarization decay model (CDDM) to calculate the degree of circular polarization () modulated by the cell allows for the derivation of the IRID on the focal plane. Experiments on LoVo cells demonstrated the refractive index of single cell can be accurately and precisely measured, with precision of 10 refractive index units (RIU). Additionally, chromatin content during the interphases (G1, S, G2) of cell cycle was recorded at 56.5%, 64.4%, and 71.5%, respectively. A significantly finer IRID can be obtained compared to the phase measurement method. This method is promising in providing a dynamic label-free intracellular substances detection method in cancer pathogenesis studies.
对活癌细胞内物质进行无标记检测仍然是癌症发病机制研究中的一个重大障碍。尽管光偏振对细胞内物质的敏感性已得到验证,但目前的研究主要集中在组织病变上,因此对单个活癌细胞内物质进行无标记检测仍是一项挑战。主要困难在于找到特定的检测方法以及相应的特征参数。以折射率作为物质的内源性标记,本研究提出了一种用于无标记活结肠癌细胞(LoVo细胞)的细胞内折射率分布(IRID)检测方法。利用圆偏振衰减模型(CDDM)计算由细胞调制的圆偏振度(),可以在焦平面上推导IRID。对LoVo细胞的实验表明,单细胞的折射率能够被精确测量,精度达到10个折射率单位(RIU)。此外,细胞周期各间期(G1、S、G2)的染色质含量分别记录为56.5%、64.4%和71.5%。与相位测量方法相比,可获得明显更精细的IRID。该方法有望为癌症发病机制研究提供一种动态的无标记细胞内物质检测方法。