Intracellular pH measurements using perfluorocarbon nanoemulsions.
Intracellular pH measurements using perfluorocarbon nanoemulsions.
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DOI：
10.1021/ja407573m
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发表时间：
2013-12-11
影响因子：
15
通讯作者：
Waggoner AS
中科院分区：
文献类型：
--
作者：
Patrick MJ;Janjic JM;Teng H;O'Hear MR;Brown CW;Stokum JA;Schmidt BF;Ahrens ET;Waggoner AS
We report the synthesis and formulation of unique perfluorocarbon (PFC) nanoemulsions enabling intracellular pH measurements in living cells via fluorescent microscopy and flow cytometry. These nanoemulsions are formulated to readily enter cells upon co-incubation and contain two cyanine-based fluorescent reporters covalently bound to the PFC molecules, specifically Cy3-PFC and CypHer5-PFC conjugates. The spectral and pH-sensing properties of the nanoemulsions where characterized in vitro and showed the unaltered spectral behavior of dyes after formulation. In rat 9L glioma cells loaded with nanoemulsion, the local pH of nanoemulsions was longitudinally quantified using optical microscopy and flow cytometry, and displayed a steady decrease in pH to a level of 5.5 over 3 hours, indicating rapid uptake of nanoemulsion to acidic compartments. Overall, these reagents enable real-time optical detection of intracellular pH in living cells in response to pharmacological manipulations. Moreover, recent approaches for in vivo cell tracking using magnetic resonance imaging (MRI) employ intracellular PFC nanoemulsion probes to track cells using 19F MRI. However, the intracellular fate of these imaging probes is poorly understood. The pH sensing nanoemulsions allow the study of the fate of the PFC tracer inside the labeled cell, which is important for understanding the PFC cell loading dynamics and nanoemulsion stability and cell viability over time.
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