Time-lapse imaging of cell death in cell culture and whole living organisms using turn-on deep-red fluorescent probes.
Time-lapse imaging of cell death in cell culture and whole living organisms using turn-on deep-red fluorescent probes.
复制标题
使用打开的深红色荧光探针对细胞培养物和整个活生物体中的细胞死亡进行延时成像。
DOI：
10.1039/c8tb01495g
复制
发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Smith,BradleyD
中科院分区：
文献类型：
--
作者：
Jarvis,TiaS;Roland,FeliciaM;Dubiak,KyleM;Huber,PaulW;Smith,BradleyD
细胞死亡是发育生物学中的一个中心过程，也是疾病状态和治疗效果的重要指标。描述了两种相关的荧光探针，它们是一种或两种吡啶二甲基胺锌（ZnDPA）配位络合物与附加的溶剂化显色苯并噻唑方酸染料的分子缀合物。这些探针被设计为靶向暴露在死亡和垂死细胞表面的阴离子磷脂，磷脂酰丝氨酸（PS）。以脂质体和红细胞血影为模型进行的一系列光谱和显微镜研究表明，具有两个ZnDPA靶向单元的探针比具有一个ZnDPA的探针产生更高的亲和力、更强的荧光“开启”效应和更好的图像对比度。这两种荧光探针都能够对培养物中的哺乳动物细胞死亡进行“无洗涤”延时显微成像。具有两个ZnDPA单元的探针用于非洲爪蟾（Xenopus laevis）（青蛙）胚胎发育过程中细胞死亡的非侵入性延时成像。体内荧光显微照片显示探针在胚胎尾部、头部和脊柱区域内积聚，这些区域在生长和成熟过程中经历退化和细胞凋亡。这些新的荧光探针可能是有用的时间分辨，非侵入性的活生物体细胞死亡过程中的活体成像。从更广泛的角度来看，应该可以利用苯并噻唑方酸染料所表现出的负溶剂化显色性来开发各种“开启”深红色荧光探针和靶向细胞表面生物标志物的材料，用于体外和体内成像。
Cell death is a central process in developmental biology and also an important indicator of disease status and treatment efficacy. Two related fluorescent probes are described that are molecular conjugates of one or two zinc dipicolylamine (ZnDPA) coordination complexes with an appended solvatochromic benzothiazolium squaraine dye. The probes were designed to target the anionic phospholipid, phosphatidylserine (PS), that is exposed on the surface of dead and dying cells. A series of spectrometric and microscopy studies using liposomes and red blood cell ghosts as models showed that the probe with two ZnDPA targeting units produced higher affinity, stronger fluorescence “turn-on” effect, and better image contrast than the probe with one ZnDPA. Both fluorescent probes enabled “no-wash” time-lapse microscopic imaging of mammalian cell death within a culture. The probe with two ZnDPA units was used for non-invasive time-lapse imaging of cell death during the development of Xenopus laevis (frog) embryos. In vivo fluorescence micrographs revealed probe accumulation within the embryo tail, head and spine regions that were undergoing regression and apoptosis during growth and maturation. These new fluorescent probes are likely to be useful for time-resolved, non-invasive in vivo imaging of cell death process in range of living organisms. From a broader perspective, it should be possible to utilize the negative solvatochromism exhibited by benzothiazolium squaraine dyes for development of various “turn-on” deep-red fluorescent probes and materials that target cell surface biomarkers for in vitro and in vivo imaging.