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Sensitive, real-time monitoring of UV-induced stress in a single, live plant cell using an optical trap

机译:使用光阱对单个活植物细胞中的紫外线诱导的胁迫进行灵敏的实时监控

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An optical trap is used to monitor bio-deleterious effects induced by ultra-violet (UV) radiation in a single, live cell. When placed in a laser optical trap, Chlamydomonas reinhardtii cells undergo rotation because of the interplay of flagellar-based forces and optical forces generated by the laser light. Such rotation is sustained by the trapped cell's flagellar action, thereby unveiling a robust and quantitative cellular assay for flagellar function. UV induces flagellar damage, leading to dose-dependent attenuation of cellular rotation. At UV doses larger than ~9 J m~(-2) rotational motion ceases. The ability of the cell to rotate in an optical trap is, therefore, a measure of the damage response of the cell. By monitoring cell rotation, we can quantify UV damage with high sensitivity, at threshold doses as low as 2 Jm~(-2). Apart from the capability of UV-damage detection, our rotation assay is also sensitive to the cellular protective responses against such damage. To illustrate this additional facet, we quantify the efficacy of ascorbic acid in combating the UV-damage response of individual cells. Upon addition of this antioxidant, there is no cessation of rotations for doses as high as 25 J m~(-2). This represents the first single-cell sensor that robustly quantifies a complex pleiotropic cellular response.
机译:光阱用于监测单个活细胞中紫外线(UV)诱导的生物有害作用。当放置在激光光阱中时,衣藻衣藻细胞会由于鞭毛力与激光产生的光学力的相互作用而发生旋转。这种旋转通过捕获的细胞的鞭毛作用得以维持,从而揭示了一种可靠且定量的鞭毛功能细胞测定法。紫外线会引起鞭毛损伤,导致细胞旋转的剂量依赖性衰减。当紫外线剂量大于〜9 J m〜(-2)时,旋转运动停止。因此,细胞在光阱中旋转的能力是细胞损伤反应的量度。通过监测细胞旋转,我们可以以2 Jm〜(-2)的阈值剂量高灵敏度地量化UV损伤。除了能够检测紫外线损伤之外,我们的旋转测定法还对针对此类损伤的细胞保护反应敏感。为了说明这个附加方面,我们量化了抗坏血酸在对抗单个细胞的紫外线损伤反应中的功效。加入抗氧化剂后,剂量高达25 J m〜(-2)时不会停止旋转。这代表了第一个可靠地量化复杂多效性细胞反应的单细胞传感器。

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