Characterization and application of Raman labels for confocal Raman microspectroscopic detection of cellular proteins in single cells. Anal Biochem 2003 Nov 15;322(2):198-207
Date
11/05/2003Pubmed ID
14596828DOI
10.1016/j.ab.2003.07.020Scopus ID
2-s2.0-0142226894 (requires institutional sign-in at Scopus site) 47 CitationsAbstract
A method using confocal Raman microspectroscopy for the detection of cellular proteins in single intact cells was developed. Two approaches were used to improve the detection of these cellular components. First, compounds with high Raman scattering were investigated for potential use as Raman labels. Raman labels were conjugated to either biomolecules or biotin and used as markers in the detection of cellular enzymes and receptors. Second, silver colloids were used to increase the surface-enhanced Raman scatter (SERS) of these Raman labels. Cresyl violet and dimethylaminoazobenzene are Raman labels that provide very sensitive SERS detection by a confocal Raman microscope with a HeNe laser at wavelength of 632.8 nm. The detection of 12-lipoxygenase and cyclooxygenase-1 in single bovine coronary artery endothelial cells and the binding of angiotensin II to its receptors in zona glomerulosa cells was demonstrated.
Author List
Nithipatikom K, McCoy MJ, Hawi SR, Nakamoto K, Adar F, Campbell WBAuthor
William B. Campbell PhD Professor in the Pharmacology and Toxicology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Angiotensin IIAnimals
Arachidonate 12-Lipoxygenase
Benzoxazines
Biotin
Cattle
Cells, Cultured
Citrates
Colloids
Coloring Agents
Coronary Vessels
Cyclooxygenase 1
Endothelium, Vascular
Fluorescent Antibody Technique
Isoenzymes
Microscopy, Confocal
Oxazines
Prostaglandin-Endoperoxide Synthases
Proteins
Receptors, Angiotensin
Sensitivity and Specificity
Silver Compounds
Spectrum Analysis, Raman
Zona Glomerulosa
p-Dimethylaminoazobenzene