Chemistry Department, Imperial College London, Exhibition Road, London SW7 2AZ, UK.
Phys Chem Chem Phys. 2012 Oct 5;14(37):12671-86. doi: 10.1039/c2cp41674c.
This article describes an emerging method for quantitative measurement and spatial imaging of microviscosity within individual domains of live cells. The method is based on fluorescence detection from small synthetic molecules termed 'molecular rotors', which are characterised by a strong response of fluorescence lifetimes or spectra to the viscosity of their immediate environment. Alongside this new method, two complementary techniques are discussed, which provide further insights into diffusion controlled processes on a microscopic scale in a biological environment. These are time resolved fluorescence anisotropy and imaging of short-lived excited state of molecular oxygen, termed 'singlet oxygen'. It is possible to utilise all three approaches for the quantitative determination of viscosity in individual organelles of live cells. Finally, it is discussed how the major advantage of molecular rotor imaging, fast signal acquisition, can be used to monitor changing viscosity during dynamic biological processes within cells, such as photoinduced cell death.
本文描述了一种新兴的方法,用于定量测量和空间成像活细胞内各个微区的微观粘度。该方法基于从小型合成分子(称为“分子转子”)的荧光检测,这些分子的荧光寿命或光谱对其周围环境的粘度具有强烈的响应。除了这种新方法,还讨论了两种互补技术,它们进一步深入了解了生物环境中微观尺度上扩散控制过程。这两种技术是时间分辨荧光各向异性和短寿命激发态氧(称为“单线态氧”)的成像。这三种方法都可用于定量测定活细胞内各个细胞器的粘度。最后,讨论了分子转子成像的主要优势,即快速信号采集,如何用于监测细胞内动态生物过程中粘度的变化,例如光诱导细胞死亡。