FLUORESCENT RHODOL DERIVATIVES - VERSATILE, PHOTOSTABLE LABELS AND TRACERS
FLUORESCENT RHODOL DERIVATIVES - VERSATILE, PHOTOSTABLE LABELS AND TRACERS
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DOI：
10.1016/0003-2697(92)90011-u
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发表时间：
1992-12-01
影响因子：
2.9
通讯作者：
PRENDERGAST, FG
中科院分区：
文献类型：
--
作者：
WHITAKER, JE;HAUGLAND, RP;PRENDERGAST, FG
制备了一系列具有化学活性的荧光罗多尔衍生物，并对其进行了评价。活性官能团包括活化酯、胺、卤代乙酰胺、可固定的酰肼衍生物、丙烯酰胺和光亲和试剂。根据不同取代基的选择，染料的吸收峰在490~550 nm之间变化，在水溶液中的消光系数一般大于50000 m−1 cm−1，发射峰在520~580 nm之间。所研究的大多数化合物的荧光寿命在3到4 ns之间。单个衍生物适合于用Ar离子激光的488和514 nm线以及汞弧灯的546 nm线激发，并与标准的荧光素和罗丹明滤光片组兼容使用。与荧光素衍生物相比，Rhodol染料的光稳定性更好，对生理范围内的pH变化不太敏感。一些例子显示在514 nm或附近的吸收峰值，这是一个对多色荧光显微镜、流式细胞术和DNA测序有用的激发波长。还制备了与四甲基罗丹明(TMR)类似物的吸收和发射峰值相似的衍生物，但在水溶液中具有更高的量子产率。与TMR衍生物相比，许多染料在水体系中具有更高的溶解度，并且与蛋白质的共轭猝灭较少。与荧光素或TMR衍生物相比，适当的取代可使其在亲水性或亲油性溶剂中的溶解度范围更广。与荧光素结合物相比，许多罗多尔荧光团的结合物具有更好的光稳定性和较低的pH敏感性，并且比TMR结合物具有更强的荧光。
A series of chemically reactive, fluorescent rhodol derivatives was prepared and evaluated. Reactive functional groups included activated esters, amines, haloacetamides, fixable hydrazide derivatives, acrylamides, and photoaffinity reagents. Depending on the choice of substituents, absorption maxima of the dyes varied from 490 to 550 nm with extinction coefficients that were generally greater than 50,000 m−1cm−1in aqueous solution and emission maxima from 520 to 580 nm. Most of the compounds investigated exhibited fluorescence lifetimes between 3 and 4 ns. Individual derivatives were suitable for excitation with the 488 and 514-nm lines of the argon ion laser and the 546-nm line of the mercury arc lamp and were compatible for use with standard fluorescein and rhodamine filter sets. The rhodol dyes were more photostable and less sensitive to pH changes in the physiological range than fluorescein derivatives. Some examples show absorption maxima at or near 514 nm, an excitation wavelength that is useful for multicolor fluorescence microscopy, flow cytometry, and DNA sequencing. Derivatives were also prepared that exhibit absorption and emission maxima similar to those of tetramethylrhodamine (TMR) analogs but with higher quantum yields in aqueous solution. A number of the dyes had higher solubilities in aqueous systems and were less quenched on conjugation to proteins than TMR derivatives. Appropriate substitution results in a wider range of solubilities in hydrophilic or lipophilic solvents than is easily accomplished with fluorescein or TMR derivatives. Conjugates of a number of the rhodol fluorophores were generally more photostable and less pH sensitive than fluorescein conjugates and more fluorescent than TMR conjugates.