期刊
NEUROPHOTONICS
卷 4, 期 1, 页码 -出版社
SPIE-SOC PHOTO-OPTICAL INSTRUMENTATION ENGINEERS
DOI: 10.1117/1.NPh.4.1.015002
关键词
extracellular space; potassium; potassium buffering; fluorescence microscopy; two-photon imaging; fluorescent indicator; nanotechnology
资金
- Swiss National Science Foundation [31003a_159513]
- Swiss National Science Foundation (SNF) [31003A_159513] Funding Source: Swiss National Science Foundation (SNF)
Neuronal activity results in the release of K+ into the extracellular space (ECS). Classically, measurements of extracellular K+ ([K+](o)) are carried out using K+-sensitive microelectrodes, which provide a single point measurement with undefined spatial resolution. An imaging approach would enable the spatiotemporal mapping of [K+](o). Here, we report on the design and characterization of a fluorescence imaging-based K+-sensitive nanosensor for the ECS based on dendrimer nanotechnology. Spectral characterization, sensitivity, and selectivity of the nanosensor were assessed by spectrofluorimetry, as well as in both wide-field and two-photon microscopy settings, demonstrating the nanosensor efficacy over the physiologically relevant ion concentration range. Spatial and temporal kinetics of the nanosensor responses were assessed using a localized iontophoretic K+ application on a two-photon imaging setup. Using acute mouse brain slices, we demonstrate that the nanosensor is retained in the ECS for extended periods of time. In addition, we present a ratiometric version of the nanosensor, validate its sensitivity in brain tissue in response to elicited neuronal activity and correlate the responses to the extracellular field potential. Together, this study demonstrates the efficacy of the K+-sensitive nanosensor approach and validates the possibility of creating multimodal nanosensors. (C) The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.
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