Wellman Center for Photomedicine, Harvard Medical School, Massachusetts General Hospital, 149 13th Street, Charlestown, Massachusetts, 02129, USA.
Harvard-MIT Division of Health Sciences and Technology, 77 Massachusetts Avenue E25-518, Cambridge, Massachusetts, 02139, USA.
Sci Rep. 2019 Apr 5;9(1):5713. doi: 10.1038/s41598-019-42165-2.
A central challenge in cancer biology is the identification, longitudinal tracking, and -omics analysis of specific cells in vivo. To this aim, photoconvertible fluorescent dyes are reporters that are characterized by a set of excitation and emission spectra that can be predictably altered, resulting in a distinct optical signature following irradiation with a specific light source. One such dye, DiR, is an infrared fluorescent membrane probe that can irreversibly undergo such a switch. Here, we demonstrate a method using DiR for the spatiotemporal labeling of specific cells in the context of cancer cell monolayer cultures, 3D tumor spheroids, and in vivo melanoma xenograft models to monitor the proliferation of cellular subpopulations of interest over time. Importantly, the photoconversion process is performed in situ, supporting the pursuit of novel avenues of research in molecular pathology.
癌症生物学中的一个核心挑战是在体内鉴定、纵向跟踪和进行特定细胞的组学分析。为此，光转化荧光染料是一种报告器，其特征在于一组激发和发射光谱，可以通过用特定光源照射来可预测地改变，从而在照射后产生独特的光学特征。其中一种染料 DiR 是一种红外荧光膜探针，它可以不可逆地进行这种转变。在这里，我们展示了一种使用 DiR 的方法，用于在癌细胞单层培养物、3D 肿瘤球体和体内黑色素瘤异种移植模型中对特定细胞进行时空标记，以随时间监测感兴趣的细胞亚群的增殖。重要的是，光转化过程是在原位进行的，支持在分子病理学领域探索新的研究途径。
