亲水支架细胞培养的染色方法
method to detect cellular culture state on the hydrophilic scaffold
目的：发现一种检测亲水支架上细胞培养状态的方法。方法：将待检测细胞接种到亲水支架上进行培养，至待检测细胞在亲水支架上的生长融合率为90%-95%时，获得支架-细胞复合体系。采用吖啶橙染色液对支架-细胞复合体系进行荧光染色，并采用磷酸缓冲盐溶液对染色后的支架-细胞复合体系进行洗涤。将洗涤后的支架-细胞复合体系置于激光扫描共聚焦显微镜下，并利用绿色荧光通道和橙色荧光通道进行观察，从而检测得到亲水支架上细胞的培养状态。结果：通过这种方法，激光共聚焦显微镜下的细胞分布情况显示十分清晰，不存在亲水支架对荧光染色液的吸附所造成的干扰，使得细胞的分布及生长状态的分析更加直观及准确。结论：本研究提供的方法能够清晰、准确地检测亲水支架上细胞的分布情况及生长状态，可以对亲水支架上的细胞进行无损、实时、动态地检测。且操作简便，便于规模化推广应用。
Purpose: in present study, a method was introduced for detecting the state of cells cultured on the hydrophilic scaffold. Methods: stem cells were cultured on a hydrophilic scaffold to build a cell-scaffold complex with a confluence of 90-95%. The fluorescence staining of cell-scaffold complex was produced by acridine orange staining, and rinsed by phosphate buffer solution(PBS) to erase the overdoes acridine orange fluorescence dyes. After fluorescent staining, the cell-scaffold complex was observed under green and orange channels of confocal fluorescence microscope. Result: the state of cultured cells could be displayed clearly and precisely without the interference of unintended staining on hydrophilic scaffold.. Conclusion: by this method, The distribution and growth state of cell on the scaffold can be measured easily.
孟庆阳、皮彦斌、敖英芳
细胞生物学生物工程学生物科学研究方法、生物科学研究技术
吖啶橙染色，激光共聚焦显微镜，亲水支架
cridine orange fluorescence dyesConfocal fluorescence microscopeHydrophilic scaffold
孟庆阳,皮彦斌,敖英芳.亲水支架细胞培养的染色方法[EB/OL].(2017-05-27)[2025-08-04].http://www.paper.edu.cn/releasepaper/content/201705-1426.点此复制
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