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Author:
Date:
2015-03-05
[Abstract] Redox homeostasis is a fundamental property of living cells and responds actively to both cellular metabolism and external stimulus. The development of redox-sensitive GFP (roGFP) enables dynamic monitoring of changes in cellular redox poise (Hanson et al., 2014). When excited alternatively at 405 nm and 488 nm, these probes exhibit significant opposing shifts at the emission spectra (505-530 nm), which enables ratiometric measurement of relative redox values. A more oxidized environment results in a higher 405/488 ratio. Previously, successful application of roGFPs in leaf epidermis or root cells has been reported. Here we provide a protocol describing the application of roGFP1 imaging in growing pollen tubes by confocal laser scanning microscopy.
[摘要] 氧化还原稳态是活细胞的基本特性，并对细胞代谢和外部刺激有积极的反应。 氧化还原敏感性GFP（roGFP）的发展使得能够动态监测细胞氧化还原时间的变化（Hanson等人，2014）。 当在405nm和488nm交替激发时，这些探针在发射光谱（505-530nm）处显示出显着的相反偏移，这使得能够比较测量相对氧化还原值。 更加氧化的环境导致更高的405/488比率。 以前，已经报道了在叶表皮或根细胞中成功应用roGFP。 在这里我们提供协议描述roGFP1成像在生长花粉管的共聚焦激光扫描显微镜的应用。
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