title: Multiplex immunoassay using fluorescent-surface enhanced Raman spectroscopic dots for the detection of bronchioalveolar stem cells in murine lung.
abstract: Immunoassays using nanomaterials have been rapidly developed for the analysis of multiple biomolecules. Highly sensitive and biocompatible surface enhanced Raman spectroscopy-active nanomaterials have been used for biomolecule analysis by many research groups in order to overcome intrinsic problems of conventional immunoassays. We used fluorescent surface-enhanced Raman spectroscopic dots (F-SERS dots) to detect biomolecules in this study. The F-SERS dots are composed of silver nanoparticle-embedded silica nanospheres, organic Raman tagging materials, and fluorescent dyes. The F-SERS dots demonstrated highly sensitive, selective, and multifunctional characteristics for multiplex targeting, tracking, and imaging of cellular and molecular events in the living organism. We successfully applied F-SERS dots for the detection of three cellular proteins, including CD34, Sca-1, and SP-C. These proteins are simultaneously expressed in bronchioalveolar stem cells (BASCs) in the murine lung. We analyzed the relative expression ratios of each protein in BASCs since external standards were used to evaluate SERS intensity in tissue. Quantitative comparisons of multiple protein expression in tissue were first attempted using SERS-encoded nanoprobes. Our results suggested that immunoassays using F-SERS dots offered significant increases in sensitivity and selectivity. Such immunoassays may serve as the primary next-generation labeling technologies for the simultaneous analysis of multiple biomolecules.
authors.affiliation: College of Veterinary Medicine and Interdisciplinary Program in Nano-Science and Technology, Seoul National University, Seoul 151-742, Korea.
authors.forename: Min-Ah
authors.initials: MA
authors.lastname: Woo
authors.forename: Sang-Myung
authors.initials: SM
authors.lastname: Lee
authors.forename: Gunsung
authors.initials: G
authors.lastname: Kim
authors.forename: JongHo
authors.initials: J
authors.lastname: Baek
authors.forename: Mi Suk
authors.initials: MS
authors.lastname: Noh
authors.forename: Ji Eun
authors.initials: JE
authors.lastname: Kim
authors.forename: Sung Jin
authors.initials: SJ
authors.lastname: Park
authors.forename: Arash
authors.initials: A
authors.lastname: Minai-Tehrani
authors.forename: Se-Chang
authors.initials: SC
authors.lastname: Park
authors.forename: Yeong Tai
authors.initials: YT
authors.lastname: Seo
authors.forename: Yong-Kwon
authors.initials: YK
authors.lastname: Kim
authors.forename: Yoon-Sik
authors.initials: YS
authors.lastname: Lee
authors.forename: Dae Hong
authors.initials: DH
authors.lastname: Jeong
authors.forename: Myung-Haing
authors.initials: MH
authors.lastname: Cho
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry; D018952:Antigens, CD34 / Q000032:analysis; D000950:Antigens, Ly / Q000032:analysis; D045744:Cell Line, Tumor; D005456:Fluorescent Dyes / Q000737:chemistry*; D007118:Immunoassay / Q000379:methods*; D036341:Intercellular Signaling Peptides and Proteins; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D053758:Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000032:analysis; D011650:Pulmonary Alveoli / Q000166:cytology*; D037721:Pulmonary Surfactant-Associated Protein C; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013234:Stem Cells / Q000378:metabolism*
chemical_list: D000906:Antibodies; D018952:Antigens, CD34; D000950:Antigens, Ly; D005456:Fluorescent Dyes; D036341:Intercellular Signaling Peptides and Proteins; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D010455:Peptides; D037721:Pulmonary Surfactant-Associated Protein C; C489680:Sftpc protein, mouse