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제108회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Multiparameter Fluorescence Spectroscopy : Single Quantum Dot-Dye FRET Hybrids

2011년 8월 5일 16시 28분 28초
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금 10시 : 40분
분석화학 - Novel applications of nanotechnologies in analytical chemistry
저자 및
김학준, Carl C. Hayden1
덕성여자대학교 화학과, Korea
1Sandia National Lab, Livermore, CA, Korea
Conformational fluctuation of macromolecules such as protein, DNA, RNA in solutions is important to understanding biological functions of living body. Fluorescent dye molecules attached to the site of interest are used to report the local environment change correlated to the conformational change of the macromolecules. We have carried out experiments using a custom-built multiparameter fluorescence microscope that measures the wavelength, emission delay relative to excitation (excited-state lifetime), and chronological time (intensity) for each detected photon. By simultaneously measuring multiparamers of fluorescence from sample, we can identify single fluorophore within a multiple-fluorophore sample based on fluorescence spectrum and fully characterize the fluorescence properties of the molecule to get thorough information on its local environment by correlating multiparameters of fluorescence from sample. We have carried out a single-molecule study of a TMR-biocytin bound to a streptavidin immobilized on PEG-biotin surfaces as well as that encapsulated in a surface tethered vesicle to understand interactions between a single dye molecule and a single protein. F?rster resonance energy transfer (FRET) between donor and acceptor fluorophores is sensitive to the separation between the fluorophores, which permits us to monitor structural changes of a macromolecule on a real-time basis. As acceptor fluorophores, semiconductor quantum dots (QDs) have several advantages over organic fluorescent dyes, such as high photostability, broad absorption, and tunable, narrow emission. We have carried out multiparameter FRET measurements for single QD-Cy5 hybrids made with biotinylated QDs and Cy5 labeled streptavidin. Based on the fluorescence spectrum of QD, we were able to obtain FRET efficiencies of QD-Cy5 hybrids of desired QD in size that influence FRET efficiency in addition to the separation between two FRET pairs.