A highly uniform UV transillumination imaging system for quantitative analysis of nucleic acids and proteins
A highly uniform UV transillumination imaging system for quantitative analysis of nucleic acids and proteins
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DOI：
10.1002/pmic.200700891
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发表时间：
2008-05-01
期刊：
影响因子：
3.4
通讯作者：
Chakravarti, Deb N.
中科院分区：
文献类型：
--
作者：
Chakravarti, Bulbul;Louie, Melissa;Chakravarti, Deb N.
用于核酸和蛋白质凝胶电泳分离记录和分析的数字荧光成像广泛应用于定量生物学。大多数用于蛋白质和核酸电泳后分析的荧光染料在紫外光 (300-365 nm) 下具有显着的激发峰，因此中程紫外线 (UV-B) 成为首选的激发源。然而，由于典型的紫外透照仪缺乏均匀性，将定量 CCD 成像与紫外耦合起来很困难。这些大分子的表观量取决于凝胶带在透照器成像表面上的位置。在这里，我们报告了高度均匀的紫外线透照系统的开发和验证。使用新颖的高密度照明系统，包含形成高密度网格的单灯、电子镇流器、磷光体涂层和带通滤波器，将产生的 254 nm 光转换为 300-340 nm，在典型的 UV 透射仪中观察到 80% CV 的均匀性。该系统已用于电泳分离的核酸和蛋白质（CV
The digital fluorescent imaging for documentation and analysis of gel electrophoretic separations of nucleic acids and proteins is widely used in quantitative biology. Most fluorescent stains used in postelectrophoretic analysis of proteins and nucleic acids have significant excitation peaks with UV light (300-365 nm), making midrange UV (UV-B) as the excitation source of choice. However, coupling quantitative CCD imaging with UV is difficult due to lack of uniformity found in typical UV transilluminators. The apparent amount of those macromolecules depends on the position of the gel band on the imaging surface of the transilluminator. Here, we report the development and validation of a highly uniform UV transillumination system. Using a novel high density lighting system containing a single lamp formed into a high density grid, an electronic ballast, a phosphor coating, and a bandpass filter to convert 254 nm light produced to 300-340 nm, uniformity of 80% CV observed in typical UV transilluminators. This system has been used for the quantitative analysis of electrophoretically separated nucleic acids and proteins (CV