WGA-QD probe-based AFM detects WGA-binding sites on cell surface and WGA-induced rigidity alternation.
A strategy involving the conjugation of fluorescent quantum dot (QD) with wheat germ agglutinin (WGA) acting as fluorescent and topographic probes prior to cell surface staining is developed for fluorescence microscopy and atomic force microscopy (AFM). This strategy provided at least two advantages: (a) an amplified fluorescence of WGA-QD aggregates, strongly resistant to photobleaching, ensures repeated/real-time observations of the probe-labeled cells by fluorescence microscopy; (b) the enlarged size of WGA-QD probe makes it possible for labeled WGA to be distinguished from other membrane proteins by AFM. Here, the random distribution of WGA-binding sites on non-crosslinked cells and the uneven or polarized reorganization due to WGA-induced crosslinking on cell surfaces were studied using AFM-detectable WGA-QD probe. Moreover, we developed a method to rapidly detect the WGA-induced rigidity alternation of the whole cells, which is efficient and has the potentiality of being developed to a useful tool in clinical diagnosis.
Wang
Xiaoping
X
Department of Anesthesiology, The First Affiliated Hospital, Jinan University, Guangzhou 510632, China.
He
Dongmei
D
Cai
Jiye
J
Chen
Tongsheng
T
Zou
Feiyan
F
Li
Yalan
Y
Wu
Yangzhe
Y
Chen
Zheng W
ZW
Chen
Yong
Y
D000538:Aluminum Silicates / Q000737:chemistry; D001665:Binding Sites; D019921:Biotinylation; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods*; D018613:Microscopy, Confocal; D045663:Quantum Dots; D019809:Streptavidin / Q000737:chemistry; D014909:Wheat Germ Agglutinins / Q000737:chemistry*
D000538:Aluminum Silicates; D005456:Fluorescent Dyes; D014909:Wheat Germ Agglutinins; D019809:Streptavidin; C011934:mica