Polarization sensing with visual detection

被引:25
作者
Gryczynski, I [1 ]
Grycsynski, Z [1 ]
Lakowicz, JR [1 ]
机构
[1] Univ Maryland, Sch Med, Dept Biochem & Mol Biol, Ctr Fluorescence Spect, Baltimore, MD 21201 USA
关键词
D O I
10.1021/ac981301i
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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 degrees, 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.
引用
收藏
页码:1241 / 1251
页数:11
相关论文
共 63 条
[51]  
SZMACINSKI H, 1997, BIOCHIM BIOPHYS ACTA, V1383, P151
[52]  
Terpetschnig E, 1997, METHOD ENZYMOL, V278, P295
[53]   FLUORESCENCE POLARIZATION IMMUNOASSAY OF A HIGH-MOLECULAR-WEIGHT ANTIGEN BASED ON A LONG-LIFETIME RU-LIGAND COMPLEX [J].
TERPETSCHNIG, E ;
SZMACINSKI, H ;
LAKOWICZ, JR .
ANALYTICAL BIOCHEMISTRY, 1995, 227 (01) :140-147
[54]  
Thompson R. B., 1994, T FLUOR SP, P151
[55]  
THOMPSON RB, 1997, SPIE P, V2980, P582
[56]   Glucose sensor for low-cost lifetime-based sensing using a genetically engineered protein [J].
Tolosa, L ;
Gryczynski, I ;
Eichhorn, LR ;
Dattelbaum, JD ;
Castellano, FN ;
Rao, G ;
Lakowicz, JR .
ANALYTICAL BIOCHEMISTRY, 1999, 267 (01) :114-120
[57]  
TSIEN RY, 1989, METHOD CELL BIOL, V30, P127
[58]   SYNTHESIS AND CHARACTERIZATION OF IMPROVED CHLORIDE-SENSITIVE FLUORESCENT INDICATORS FOR BIOLOGICAL APPLICATIONS [J].
VERKMAN, AS ;
SELLERS, MC ;
CHAO, AC ;
LEUNG, T ;
KETCHAM, R .
ANALYTICAL BIOCHEMISTRY, 1989, 178 (02) :355-361
[60]   Photoactive ruthenium(II) cyclodextrins responsive to guest binding [J].
Weidner, S ;
Pikramenou, Z .
CHEMICAL COMMUNICATIONS, 1998, (14) :1473-1474