Highly sensitive rare cell detection based on quantum dot probe fluorescence analysis.
This study presents an efficient and sensitive method for detecting rare cells without cell culture, in which cells are analyzed quantitatively using quantum dots (QDs) as a fluorescent probe. By the conjugation of QDs with cells, the biotin-streptavidin reaction functions as a bridge to connect QDs and cells. The cells can be quantified based on the correlation of the QD fluorescence intensity with the cell population. Non-specific adsorption and cross-reaction of QD625-streptavidin on T cell membrane are neglected by reacting with biotin anti-human CD3 and mixing with red blood cell, respectively. Additionally, the photo-activation period and pH can be controlled to enhance the fluorescence of cell populations, which increases linearly with the number of T cells from 40 to 100,000, not only in a single T cell line but also in mixing with a total of 10(6) red blood cells. Moreover, the specific T cells can be detected in less than 15 min, even though rare specific cells may number only 40 cells. Among the advantages, the proposed system for detecting rare cells include simplicity of preparation, low cost, rapid detection, and high sensitivity, all of which can facilitate the detection of circulating tumor cells in early stages of diagnosis or prognosis.
Hsieh
Yi-Heui
YH
National Synchrotron Radiation Research Center, 101 Hsin-Ann Road, Hsinchu Science Park, Hsinchu, 30076, Taiwan, Republic of China.
Liu
Shih-Jen
SJ
Chen
Hsin-Wei
HW
Lin
Yao-Kwang
YK
Liang
Keng S
KS
Lai
Lee-Jene
LJ
D015374:Biosensing Techniques / Q000191:economics / Q000379:methods*; D019921:Biotinylation; D017252:CD3 Complex / Q000276:immunology; D004912:Erythrocytes / Q000166:cytology; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry; D006801:Humans; D019169:Jurkat Cells; D045663:Quantum Dots; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000737:chemistry; D013601:T-Lymphocytes / Q000166:cytology*
D017252:CD3 Complex; D005456:Fluorescent Dyes; D019809:Streptavidin