Multifunctional aggregation-based fluorescent probe for visualizing intracellular calcium dynamic fluctuations
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RESEARCH PAPER
Multifunctional aggregation-based fluorescent probe for visualizing intracellular calcium dynamic fluctuations Hang Su 1 & Ping Ping Hu 1
&
Xiao Li Jiang 1 & Tong Tong Wu 1 & Xiao Yong Tong 1
Received: 12 May 2020 / Revised: 21 July 2020 / Accepted: 28 July 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Calcium ion (Ca2+) is an indispensable second messenger in living organisms. The impaired Ca2+ handling can induce many diseases. In this paper, we developed a simple and effective method to encapsulate a coumarin-based Ca2+ probe ((E)-2-hydroxyN′-((7-hydroxy-2-oxo-2H-chromen-8-yl)methylene)-2-phenylacetohydrazide, CPM) into nanoparticles (NPs), and CPM NPs with blue fluorescence were obtained, whose maximum excitation and maximum emission wavelengths were characterized at 365 nm and 450 nm, respectively. The CPM NPs show significant fluorescence enhancement toward Ca2+ over other metal ions, with a limit of determination (LOD) of 0.04 μM. To optimize the optical property of the NPs, CPM and curcumin, which were introduced as the Förster resonance energy transfer (FRET) donor and acceptor, respectively, were co-encapsulated, and bright green CPM@Cur NPs with large stokes shift and narrow emission band width were constructed. Due to their low cytotoxicity and excellent stability, CPM NPs and CPM@Cur NPs were further successfully used to discriminate the primary aortic smooth muscle cells isolated from mice with abnormal Ca2+ homeostasis from their littermate controls. It is worth noting that CPM@Cur NPs exhibit stronger fluorescence signal and diminished background interference, which make them have great potential in the Ca2+ monitoring during biological processes. This strategy opens a new way to synthesize NPs with high brightness and has a potential application prospect in composite sensing and intracellular imaging. Keywords Ca2+ sensing . Förster resonance energy transfer . CPM@Cur NPs . Bio-imaging . Sarco/endoplasmic reticulum Ca2+ ATPase
Introduction Calcium ions (Ca2+), as an indispensable structural component of living organisms, play a key regulation role during many cellular processes, almost ranging from embryogenesis to neural function [1, 2]. In normal individuals, the concentration of cytoplasmic Ca2+ is usually maintained at a consistent level of about 1.4 mM [3], while the [Ca2+] fluctuates Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00216-020-02851-x) contains supplementary material, which is available to authorized users. * Ping Ping Hu [email protected] * Xiao Yong Tong [email protected] 1
Chongqing Key Laboratory of Natural Product Synthesis and Drug Research, School of Pharmaceutical Sciences, Chongqing University, Chongqing 401331, China
dynamically owing to the activation of Ca2+ influx and efflux pathways, under physiological conditions or external stimulation [1]. For example, in a normal resting state, the intracellular [Ca2+] is about 0.1–0.2 μM, which changes to about 2.0– 10 μM in
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