High-Contrast Fluorescence Microscopy for a Biomolecular Analysis Based on Polarization Techniques Using an Optical Interference Mirror Slide

Biosensors

Mitsuru Yasuda²  Takuo Akimoto¹ 

[1] School of Bioscience and Biotechnology, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan; E-Mail

关键词: fluorescence microscopy;    fluorescence enhancement;    biochip;   

DOI  :  10.3390/bios4040513

来源: mdpi

【 摘 要 】

Fluorescence microscopy with an improved contrast for fluorescence images is developed using an optical interference mirror (OIM) slide, which can enhance the fluorescence from a fluorophore as a result of the double interference of the excitation light and emission light. To improve the contrast of a fluorescence image using an OIM slide, a linearly-polarized excitation light was employed, and the fluorescence emission polarized perpendicular to the polarization of the excitation light was detected. The image contrast with this optical system was improved 110-fold for rhodamine B spotted on the OIM, in comparison with a glass slide using a general fluorescence microscopy optical system. Moreover, a 24-fold improvement of the image contrast was achieved for the detection of Cy3-labeled streptavidin bound to immobilize biotin.

【 授权许可】

CC BY   
© 2014 by the authors; licensee MDPI, Basel, Switzerland.