Quantitative confocal spectroscopy — rectifying the limitations of single molecule detection

Author

Liu, Kelvin J. ; Wang, Tza-Huei

Author_Institution

Dept. of Biomed. Eng., Johns Hopkins Univ., Baltimore, MD

fYear

2008

fDate

6-9 Jan. 2008

Firstpage

1189

Lastpage

1192

Abstract

Quantitative confocal spectroscopy (QCS) is a new implementation of single molecule detection that allows highly quantitative and accurate analysis of single fluorescent molecules. It rectifies the limitations of standard confocal spectroscopy by significantly enhancing mass detection efficiency and observation volume uniformity while preserving high signal to noise ratio and single molecule sensitivity. This is accomplished by incorporation of a cylindrical lens and a novel microfabricated confocal aperture to perform a 1-D expansion of the observation volume such that it spans the entire cross-section of a typical microchannel. First, QCS is theoretically compared to standard SMD using a combination of optical modeling and Monte Carlo simulation. Then the performance of QCS is experimentally validated against standard SMD using PicoGreen stained pBR322 DNA.

Keywords

DNA; biomedical optical imaging; fluorescence spectroscopy; micro-optics; 1D expansion; Monte Carlo simulation; PicoGreen stained pBR322 DNA; confocal aperture; cylindrical lens; enhanced mass detection efficiency; observation volume uniformity; optical modeling; quantitative confocal spectroscopy; single fluorescent molecules; single molecule detection; Apertures; Biomedical engineering; DNA; Diffraction; Fluorescence; Lenses; Mass spectroscopy; Shape; Signal to noise ratio; USA Councils; confocal spectroscopy; mass detection efficiency; single molecule;

fLanguage

English

Publisher

ieee

Conference_Titel

Nano/Micro Engineered and Molecular Systems, 2008. NEMS 2008. 3rd IEEE International Conference on

Conference_Location

Sanya

Print_ISBN

978-1-4244-1907-4

Electronic_ISBN

978-1-4244-1908-1

Type

conf

DOI

10.1109/NEMS.2008.4484530

Filename

4484530