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Labeling DNA Probes03:31

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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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Optimized Staining and Proliferation Modeling Methods for Cell Division Monitoring using Cell Tracking Dyes
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探测细胞增殖:对染料选择的考虑

Kah Teong Soh1, Joseph D Tario2, Katharine A Muirhead3

  • 1Agenus, Inc., Lexington, MA, United States; Roswell Park Comprehensive Cancer Center, Buffalo, NY, United States.

Methods in cell biology
|May 5, 2024
PubMed
概括
此摘要是机器生成的。

细胞追踪染料,包括蛋白质反应和膜合类型,对于监测细胞分裂至关重要. 蛋白质反应性染料比膜染料的损耗更快,表明分裂独立的损耗.

关键词:
细胞分裂 细胞分裂细胞追踪 细胞追踪在CellTraceTM染料中使用.在CellVue®染料中使用.染料稀释剂的稀释方法流动细胞计量流动细胞计量在PKH染料中使用PKH染料.核武器的扩散

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科学领域:

  • 细胞生物学 细胞生物学
  • 免疫学 免疫学 免疫学
  • 生物技术是生物技术.

背景情况:

  • 细胞追踪染料是用于监测细胞命运和增殖的光化合物.
  • 存在两个主要类别:蛋白质反应性染料 (例如,CFSE) 和膜间染料 (例如,PKH).
  • 有效的细胞追踪需要明亮,均的染色,而不会影响细胞功能或活力.

研究的目的:

  • 提供关于使用蛋白反应性和膜间染料标记细胞系和外围血液单核细胞的考虑.
  • 呈现优化实验和关键染色程序以减轻常见风险.
  • 为了比较蛋白质反应性和膜合性染料之间的染料损失率.

主要方法:

  • 细胞系和外围血液单核细胞的标记使用蛋白反应性和膜间染料.
  • 优化染色程序的优化. 染色程序的优化.
  • 使用双染色细胞对六天内染料损失率的比较分析.

主要成果:

  • 双染色细胞表现出类似于单染色细胞的染料损失模式.
  • 与膜间染料相比,蛋白质反应性染料显示光强度的下降速度更快.
  • 这种更快的损失表明蛋白质反应性染料的额外的分离独立染料损失机制.

结论:

  • 两种蛋白质反应性和膜间接染料都可以有效地用于细胞跟踪,适当的优化.
  • 膜间染料可能提供更稳定的长期跟踪,由于较慢,潜在的分裂依赖的损失.
  • 了解染料损失动力学对于准确解释细胞增殖和命运研究至关重要.