Dynamic [Cl(-)](i) measurement with chloride sensing quantum dots nanosensor in epithelial cells.
We have synthesized a chloride sensing quantum dots (QD) nanosensor, Cl-QD, for the dynamic measurements of chloride ion concentration in the millimolar range, a sensitivity that is applicable to most physiological intracellular chloride ion concentration ([Cl(-)](i)) measurements in epithelial cells. The Cl-QD is synthesized by conjugating an anion receptor, 1-(2-mercapto-ethyl)-3-phenyl-thiourea (MEPTU) to a water soluble CdSe/ZnS QD at an emission wavelength of 620 nm. Upon binding of chloride ions to the Cl-QD, a photo-induced electron transfer mechanism caused the fluorescence of the QD to quench. This resulted in an inversely proportional relationship between the chloride ion concentration and the fluorescence intensity of the Cl-QD. We have utilized this Cl-QD to measure [Cl(-)](i) in T84 and CF-PAC cultured cells, with either the C1C-2 or CFTR chloride channels being manipulated by pharmacological chloride channel activators and inhibitors. Activations of C1C-2 and CFTR chloride channels in T84 by the respective lubiprostone and genistein caused predictive increases in the fluorescence of the Cl-QD, i.e., a decrease of [Cl(-)](i). Conversely, glibenclamide, a chloride channel inhibitor, applied to the CF-PAC cells caused a predictable decrease in the fluorescence of Cl-QD due to the increase of [Cl(-)](i). These are the first data in using QD-based chloride ion sensors for dynamic measurements of intracellular chloride ion concentrations in epithelial cells.
Wang
Yuchi
Y
BioTechPlex Corporation, 1205 Linda Vista Drive Suite A, San Marcos, CA 92078, USA. ywang@biotechplex.com
Mao
Hua
H
Wong
Lid B
LB
D015374:Biosensing Techniques / Q000379:methods*; D019187:Cadmium Compounds / Q000737:chemistry; D002460:Cell Line; D018118:Chloride Channels / Q000737:chemistry; D002712:Chlorides / Q000032:analysis*; D004847:Epithelial Cells / Q000737:chemistry*; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology; D036103:Nanotechnology / Q000379:methods*; D045663:Quantum Dots; D018036:Selenium Compounds / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013440:Sulfides / Q000737:chemistry; D017967:Zinc Compounds / Q000737:chemistry
D019187:Cadmium Compounds; D018118:Chloride Channels; D002712:Chlorides; D018036:Selenium Compounds; D013440:Sulfides; D017967:Zinc Compounds; C058667:cadmium selenide; C031238:zinc sulfide