基于激光共聚焦显微镜研究细胞核标识技术

Nucleus Identification Technology Based on Laser Confocal Microscopy

本文利用激光扫描共聚焦显微镜研究实验室常用细胞核标记方法,对细胞成像相关实验技术进行进一步优化。首先用DNA荧光标记法、组蛋白荧光标记法以及微分干涉术(DIC)分别对贴壁培养的细胞进行成像;然后结合核仁蛋白标记和核内非编码RNA杂交标记对细胞核结构进行解析,进一步挖掘常规核染色标记所能带来的深度信息;最后针对实验平台日常细胞影像实验需求进行研究,对几类常用核标识技术的优势与不足进行讨论。基于影像实验数据,可以发现染色质相关荧光标记不仅仅可用于细胞核标识,其展示的核内染色质密度分布同时暗含了细胞核的亚结构信息,我们在此予以初步实验论述。另外,利用共聚焦DIC成像同样能清晰识别细胞核及其内部核仁结构,因而DIC在一定程度上与染色质荧光标记成像是等效的。总之,本文对常用几种细胞核标识技术进行了全面介绍,有望为细胞成像相关实验的研究与分析提供新的思路。

In this paper, we use the laser scanning confocal microscopy research laboratory commonly used nuclear labeling method to further opti-mize the experimental technology related to cell imaging. First, the adherent cultured cells were imaged by DNA fluorescent labeling, histone fluorescent labeling and differential interferometry (DIC). Then combined with nucleolar protein markers and nuclear non-coding RNA hybrid markers to analyze the nuclear structure, and further explore the depth information brought by convention-al nuclear staining markers;finally, according to the needs of daily cell imaging experiments on the experimental platform, the advantages and disadvantages of several types of commonly used nu-clear labeling techniques are discussed. Based on the image experimental data, it can be found that chromatin-related fluorescent markers can not only be used for nuclear identification, but also the chromatin density distribution in the nucleus implies the substructure information of the nucleus. In addition, confocal DIC imaging can also clearly identify the nucleus and its internal nucleolar structure, so DIC is equivalent to chromatin fluorescent labeling imaging to a certain extent. In con-clusion, this paper provides a comprehensive introduction to several commonly used nuclear la-beling techniques, which is expected to provide new ideas for the research and analysis of cell im-aging-related experiments.