Dual-color prism-type TIRFM system for direct detection of single-biomolecules on nanoarray biochips

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Title
Dual-color prism-type TIRFM system for direct detection of single-biomolecules on nanoarray biochips
Author(s)
S Lee; Bong Hyun Chung; S H Kang
Bibliographic Citation
Current Applied Physics, vol. 8, no. 6, pp. 700-705
Publication Year
2008
Abstract
This study examined the applicability of a prism-type simultaneous dual-color total internal reflection fluorescence microscopy (TIRFM) system for the simultaneous detection of nano biomolecules on nanoarray biochips at the single-molecule level. The dual-color TIRFM system with two individual laser beams and a high-sensitivity camera was used for the simultaneous dual-color detection of two different nano beads (i.e. 20 nm yellow-green and crimson fluorescent FluoSpheres beads), and single-protein molecules labeled with different fluorescent dyes (i.e. actin from rabbit muscle conjugated with Alexa Fluor® 488 and Alexa Fluor® 633 goat anti-rabbit IgG) without a time-delay and the need to move the sample. When this system was applied to two different single-protein molecules labeled with different fluorescent dyes on the GPTS/CHI/GA-modified glass nanoarray chip, the full images of the biomolecules at the single-molecule level were obtained simultaneously in two different colors using a Dual-View™. The dual-color TIRFM system is quite suitable for the biological imaging at the single-molecule level on nanoarray biochips. This study provides a benchmark for directly monitoring the interactions and detecting the colocalization of two different nano biomolecules, and can be applied to the development of a nanoarray biochip at the single-molecule level.
Keyword
Dual-color TIRFMNanoarray biochipSingle-molecule detection
ISSN
1567-1739
Publisher
Elsevier
DOI
http://dx.doi.org/10.1016/j.cap.2007.04.041
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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