Multiplex immunoassay using fluorescent-surface enhanced Raman spectroscopic dots for the detection of bronchioalveolar stem cells in murine lung.
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.
Woo
Min-Ah
MA
College of Veterinary Medicine and Interdisciplinary Program in Nano-Science and Technology, Seoul National University, Seoul 151-742, Korea.
Lee
Sang-Myung
SM
Kim
Gunsung
G
Baek
JongHo
J
Noh
Mi Suk
MS
Kim
Ji Eun
JE
Park
Sung Jin
SJ
Minai-Tehrani
Arash
A
Park
Se-Chang
SC
Seo
Yeong Tai
YT
Kim
Yong-Kwon
YK
Lee
Yoon-Sik
YS
Jeong
Dae Hong
DH
Cho
Myung-Haing
MH
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*
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