期刊
ACS APPLIED MATERIALS & INTERFACES
卷 14, 期 2, 页码 2578-2586出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsami.1c19926
关键词
FLIM; fluorescence lifetime probe; intracellular pH quantification; histidine-based nanoparticle; quantum dot
资金
- Universidad de CastillaLa Mancha [2019-GRIN-27175]
- Diputacion de Albacete [DIPUAB-16-GARZONRUIZ]
- Universidad de Castilla-La Mancha
- Universidad de Castilla-La Mancha [2019/5964]
- Junta de Comunidades de Castilla-La Mancha [2018/15132]
This study investigates the use of two nanoparticles for quantitative pH measurements in live cells using FLIM. One nanoparticle (CdSe/ZnS-A) has a tendency to aggregate and shows nonlinear pH sensitivity in a complex medium, while the other nanoparticle (CdSe/ZnS-PH) exhibits chemical stability, low toxicity, efficient uptake in cells, and good performance as an FLIM probe.
The use of two nanoparticles for quantitative pH measurements in live cells by means of fluorescence lifetime imaging microscopy (FLIM) is investigated here. These nanoparticles are based on CdSe/ZnS quantum dots (QDs), functionalized with N-acetylcysteine (CdSe/ZnS-A) and with a small peptide containing D-penicillamine and histidine (CdSe/ZnS-PH). CdSe/ZnS-A has tendency to aggregate and nonlinear pH sensitivity in a complex medium containing salts and macromolecules. On the contrary, CdSe/ZnS-PH shows chemical stability, low toxicity, efficient uptake in C3H10T1/2 cells, and good performance as an FLIM probe. CdSe/ZnS-PH also has key advantages over a recently reported probe based on a CdSe/ZnS QD functionalized with D-penicillamine (longer lifetimes and higher pH-sensitivity). A pH(+/- 2s) of 6.97 +/- 0.14 was determined for C3H10T1/2 cells by FLIM employing this nanoprobe. In addition, the fluorescence lifetime signal remains nearly constant for C3H10T1/2 cells treated with CdSe/ZnS-PH for 24 h. These results show the promising applications of this nanoprobe to monitor the intracellular pH and cell state employing the FLIM technique.
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