Metal plasmon-coupled fluorescence imaging and label free coenzyme detection in cells.
Flavin adenine dinucleotide (FAD) is a key metabolite in cellular energy conversion. Flavin can also bind with some enzymes in the metabolic pathway and the binding sites may be changed due to the disease progression. Thus, there is interest on studying its expression level, distribution, and redox state within the cells. FAD is naturally fluorescent, but it has a modest extinction coefficient and quantum yield. Hence the intrinsic emission from FAD is generally too weak to be isolated distinctly from the cellular backgrounds in fluorescence cell imaging. In this article, the metal nanostructures on the glass coverslips were used as substrates to measure FAD in cells. Particulate silver films were fabricated with an optical resonance near the absorption and the emission wavelengths of FAD which can lead to efficient coupling interactions. As a result, the emission intensity and quantum yield by FAD were greatly increased and the lifetime was dramatically shortened resulting in less interference from the longer lived cellular background. This feature may overcome the technical limits that hinder the direct observation of intrinsically fluorescent coenzymes in the cells by fluorescence microscopy. Fluorescence cell imaging on the metallic particle substrates may provide a non-invasive strategy for collecting the information of coenzymes in cells.
Center for Fluorescence Spectroscopy, University of Maryland School of Medicine, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201, USA. jian@cfs.bioment.umaryland.edu
Jian
J
Zhang
Yi
Y
Fu
Ge
G
Li
Richard Y
RY
Zhao
D003067:Coenzymes / Q000032:analysis; D005182:Flavin-Adenine Dinucleotide / Q000032:analysis*; D005453:Fluorescence; D006367:HeLa Cells; D006801:Humans; D053768:Metal Nanoparticles; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods*; D020349:Surface Plasmon Resonance / Q000379:methods*
D003067:Coenzymes; D005182:Flavin-Adenine Dinucleotide