Gryczynski I, Gryczynski Z, Lakowicz J R
Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore 21201, USA.
Anal Chem. 1999 Apr 1;71(7):1241-51. doi: 10.1021/ac981301i.
We describe a new approach to fluorescence sensing which relies on visual determination the polarization. The sensing device consists of a fluorescent probe, which changes intensity in responses to the analyte, and an oriented fluorescent film, which is not affected by the analyte. An emission filter is selected to observe the emission from both the film and the sensing fluorophore. Changes in the probe intensity result in changes in the polarization of the combined emission from the sensor and reference. The degree of polarization can be detected visually using a dual polarizer with adjacent sections oriented orthogonally to each other. The emission passing through the dual polarizer is viewed with a second analyzing polarizer. This analyzer is rotated manually to yield equal intensities from both sides of the dual polarizer. This approach was used to measure the concentration of RhB in intralipid and to measure pH using 6-carboxyfluorescein. The analyzer angle is typically accurate to 1 degree, providing pH values accurate to +/- 0.1 pH unit at the midpoint of the titration curve. We also describe a method of visual polarization sensing that does not require an oriented film and that can use the same fluorophore for the sample and reference. These approaches to visual sensing are generic and can be applied to a wide variety of analytes for which fluorescent probes are available. Importantly, the devices are simple, with the only electronic component being the light source.
我们描述了一种新的荧光传感方法,该方法依赖于通过视觉确定偏振。传感装置由一个荧光探针和一个取向荧光膜组成,荧光探针会根据分析物改变强度,而取向荧光膜不受分析物影响。选择一个发射滤光片来观察膜和传感荧光团的发射。探针强度的变化会导致传感器和参考物组合发射的偏振发生变化。可以使用相邻部分相互正交取向的双偏振器通过视觉检测偏振度。透过双偏振器的发射光用第二个分析偏振器进行观察。手动旋转这个分析器,使双偏振器两侧的强度相等。该方法用于测量脂质乳剂中罗丹明B的浓度以及使用6-羧基荧光素测量pH值。分析器角度通常精确到1度,在滴定曲线中点处提供精确到±0.1 pH单位的pH值。我们还描述了一种无需取向膜的视觉偏振传感方法,该方法可对样品和参考物使用相同的荧光团。这些视觉传感方法具有通用性,可应用于有荧光探针可用的多种分析物。重要的是,这些装置很简单,唯一的电子元件是光源。