Title :
Approaches for stoichiometry and distance determination of nanometer bio-complex by dual-channel single molecule imaging
Author :
Zhang, Hui ; Shu, Dan ; Browne, Mark ; Guo, Peixuan
Author_Institution :
Dept. of Biomed. Eng., Univ. of Cincinnati, Cincinnati, OH
Abstract :
A dual-channel imaging system with single fluorophore sensitivity was assembled in this lab. Inclusion of an integrated laser combiner was introduced to facilitate simultaneous dual-channel imaging. The imaging system has been applied to study the structure, stoichiometry, distance and function of the phi29 DNA packaging motor. Approaches including single molecule photobleaching, single molecule FRET and binomial distribution quantification were carried out to clarify the stoichiometry and distance of pRNA on the biologically active packaging motor. The results were statistically analyzed to access the copy number of pRNA, and the distance constraint was used to verify the 3D structure of the computer model of phi29 DNA packaging motor.
Keywords :
DNA; binomial distribution; bio-optics; biochemistry; biological techniques; molecular biophysics; optical saturable absorption; stoichiometry; binomial distribution quantification; distance determination; dual-channel single molecule imaging; laser combiner; nanometer bio-complex; pRNA; phi29 DNA packaging motor; photobleaching; single molecule FRET; stoichiometry; Biomedical optical imaging; Fiber lasers; Fluorescence; Laser beams; Nanobioscience; Optical imaging; Optical noise; Optical sensors; Packaging; Photobleaching;
Conference_Titel :
Life Science Systems and Applications Workshop, 2009. LiSSA 2009. IEEE/NIH
Conference_Location :
Bethesda, MD
Print_ISBN :
978-1-4244-4292-8
Electronic_ISBN :
978-1-4244-4293-5
DOI :
10.1109/LISSA.2009.4906725
