| Improving the sensitivity of protein microarray by evanescent-field-induced fluorescence |
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| 作者姓名： | 王立强 陆祖康 |
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| 作者单位： | State Key Lab of Modern Optical Instrumentation Zhejiang University，State Key Lab of Modern Optical Instrumentation Zhejiang University Hangzhou 310027，China，Hangzhou 310027，China |
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| 摘 要： | To improve the sensitivity of protein microarray, a prism surface replaces the surface of the common microscope slide. The protein targets arrayed on the surface are hybridized and labelled by fluorescent probes. Evanescent excitation occurs when the convergent laser reaches the surface, and a photomultiplier tube detects the emitted fluorescent signal. A two-dimensional actuator scans the whole surface to achieve planar laser excitation and fluorescence collection. The penetration depth of the evanescent field into the protein targets is only some hundred nanometers and can be controlled by different incident angle of the laser beam, so the undesired background signals are reduced dramatically and the detection sensitivity is improved by a factor of 50 to 100 comparing to confocal excitation. This approach can detect low abundance analytes without signal amplification.
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| 关 键 词： | 蛋白质 微观排列 荧光性 敏感度 |
| 收稿时间： | 2005-01-28 |
| 修稿时间： | 2005-04-05 |
Improving the sensitivity of protein microarray by evanescent-field-induced fluorescence |
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| Wang Li-qiang,Lu Zu-kang.Improving the sensitivity of protein microarray by evanescent-field-induced fluorescence[J].Journal of Zhejiang University Science,2005,6(7):623-626. |
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| Authors: | Wang Li-qiang Lu Zu-kang |
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| Institution: | (1) State Key Lab of Modern Optical Instrumentation, Zhejiang University, 310027 Hangzhou, China |
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| Abstract: | To improve the sensitivity of protein microarray, a prism surface replaces the surface of the common microscope slide.The protein targets arrayed on the surface are hybridized and labelled by fluorescent probes. Evanescent excitation occurs when the convergent laser reaches the surface, and a photomultiplier tube detects the emitted fluorescent signal. A two-dimensional actuator scans the whole surface to achieve planar laser excitation and fluorescence collection. The penetration depth of the evanescent field into the protein targets is only some hundred nanometers and can be controlled by different incident angle of the laser beam, so the undesired background signals are reduced dramatically and the detection sensitivity is improved by a factor of 50 to 100 comparing to confocal excitation. This approach can detect low abundance analytes without signal amplification. |
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| Keywords: | Protein microarray Evanescent excitation Sensitivity Total reflection |
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