A low affinity nanoparticle based fluorescent ratiometric probe for the determination of Zn(ii) concentrations in living cells (original) (raw)

ICPBCZin: A red emitting ratiometric fluorescent indicator with nanomolar affinity for Zn 2+ ions

Cell Calcium, 2008

A new fluorescent Zn2+ indicator, namely, ICPBCZin was synthesized and the spectral profile of its free and Zn2+ bound forms was studied. The newly synthesized zinc indicator incorporates as chromophore the chromeno [3′,2′:3,4]pyrido[1,2a] [1,3]benzimidazole moiety and belongs to the dicarboxylate-type of zinc probes. The compound is excited with visible light, exhibits high selectivity for zinc in the presence of calcium and other common biological ions, and its Zn2+ dissociation constant is 4.0 nM. Fluorescence spectra studies of ICPBCZin indicated a clear shift in its emission wavelength maxima upon Zn2+ binding, as it belongs to the class of Photoinduced Charge Transfer (PCT) indicators, along with changes in fluorescence intensity that enable the compound to be used as a ratiometric, visible-excitable Zn2+ probe.

Synthesis and characterization of new selective Zn2+ fluorescent probes for functionalization: in vitro Cell imaging applications

Tetrahedron, 2013

Herein the synthesis and characterization of new, lipophilic highly Zn 2þ-selective fluorescent probes are reported. High affinity for zinc (K d 1.1e8.0 nM) over other biologically relevant metals and mixtures of metals was observed. Excitation at 360 nm afforded an emission spectrum with maximum at 530 nm for the zinc bound complex. The linear relationship between fluorescence intensity and zinc concentration indicates that FZnA-probes can be used for quantification. The probes have been synthesized in 28e45% overall yield and the feasibility for further functionalization with biologically relevant side chains has been demonstrated. In vitro studies using PC12 cells and 10 mM of one of the novel probes (FZnA-Ada) visualized endogenous labile Zn 2þ after 45 min incubation time.

Intracellular imaging of zinc ion in living cells by fluorescein based organic nanoparticles

Sensors and Actuators B: Chemical, 2018

 Novel fluorescein based nano probe (FBNPs) was designed and synthesized  Selective fluorescence enhancement of FBNPs by Zn 2+ ion  Fluorescence enhancement is an account for chelation of FBNPs and Zn 2+ in 1:1 stoichiometry.  Lower detection limit for Zn 2+ = 0.0011 µg/mL in an aqueous medium.  FBNPs exhibits excellent hydrophilicity, biocompatibility, and non-toxicity for intracellular detection of Zn 2+ in MDA-MB-231 breast cancer cell line.

Measuring zinc in living cells

Cell Calcium, 2002

New fluorescent indicators with nanomolar to micromolar affinities for Zn 2+ have been synthesized in wavelengths from UV to the far red. The UV light-excited indicators are ratiometric. The visible wavelength indicators are non-ratiometric and exhibit large and pH-independent fluorescence increases with increasing zinc concentrations, with little to no sensitivity to physiologically relevant Ca 2+ concentrations. Experiments in neuronal and non-neuronal cell cultures show the new indicators to retain their sensitivity to and selectivity for zinc after conversion to cell-permeable forms.

Detecting Zn(II) Ions in Live Cells with Near-Infrared Fluorescent Probes

Molecules, 2019

Two near-infrared fluorescent probes (A and B) containing hemicyanine structures appended to dipicolylamine (DPA), and a dipicolylamine derivative where one pyridine was substituted with pyrazine, respectively, were synthesized and tested for the identification of Zn(II) ions in live cells. In both probes, an acetyl group is attached to the phenolic oxygen atom of the hemicyanine platform to decrease the probe fluorescence background. Probe A displays sensitive fluorescence responses and binds preferentially to Zn(II) ions over other metal ions such as Cd2+ ions with a low detection limit of 0.45 nM. In contrast, the emission spectra of probe B is not significantly affected if Zn(II) ions are added. Probe A possesses excellent membrane permeability and low cytotoxicity, allowing for sensitive imaging of both exogenously supplemented Zn(II) ions in live cells, and endogenously releases Zn(II) ions in cells after treatment of 2,2-dithiodipyridine.

High-Affinity Ratiometric Fluorescence Probe Based on 6-Amino-2,2′-Bipyridine Scaffold for Endogenous Zn2+ and Its Application to Living Cells

Molecules, 2022

Zinc is an essential trace element involved in many biological activities; however, its functions are not fully understood. To elucidate the role of endogenous labile Zn2+, we developed a novel ratiometric fluorescence probe, 5-(4-methoxyphenyl)-4-(methylsulfanyl)-[2,2′-bipyridin]-6-amine (6 (rBpyZ)) based on the 6-amino-2,2′-bipyridine scaffold, which acts as both the chelating agent for Zn2+ and the fluorescent moiety. The methoxy group acted as an electron donor, enabling the intramolecular charge transfer state of 6 (rBpyZ), and a ratiometric fluorescence response consisting of a decrease at the emission wavelength of 438 nm and a corresponding increase at the emission wavelength of 465 nm was observed. The ratiometric probe 6 (rBpyZ) exhibited a nanomolar-level dissociation constant (Kd = 0.77 nM), a large Stokes shift (139 nm), and an excellent detection limit (0.10 nM) under physiological conditions. Moreover, fluorescence imaging using A549 human lung adenocarcinoma cells re...

A selective and sensitive fluorescence probe for imaging endogenous zinc in living cells

Tetrahedron, 2013

ABSTRACT A carboxamidoquinoline-based fluorescent Zn2+ probe 1 containing N/S/S heteroatoms as a receptor was designed and readily synthesized, which was featured by the Zn2+-induced red-shift of emission (45 nm) based on internal charge transfer (ICT) in an aqueous HEPES buffer (pH=7.4). Moreover, spectroscopic studies indicated that the composition of the complex 1-Zn2+ was 1:1, which was also confirmed by X-ray crystallography. In addition, 1H NMR titration experiment suggested that probe 1 transformed from the amide tautomer to imidic acid tautomer after binding with Zn2+. The binding of zinc with 1 was easily reversed by addition of N,N,N′,N′-tetrakis(2-picolyl)ethylenediamine (TPEN) or EDTA. Furthermore, probe 1 was successfully applied to image both exogenous and endogenous zinc in living HeLa cells.

Peptide-Based, Two-Fluorophore, Ratiometric Probe for Quantifying Mobile Zinc in Biological Solutions

ACS Chemical Biology, 2014

Small-molecule fluorescent sensors are versatile agents for detecting mobile zinc in biology. Capitalizing on the abundance of validated mobile zinc probes, we devised a strategy for repurposing existing intensity-based sensors for quantitative applications. Using solid-phase peptide synthesis, we conjugated a zinc-sensitive Zinpyr-1 derivative and a zinc-insensitive 7-hydroxycoumarin derivative onto opposite ends of a rigid P 9 K peptide scaffold to create HcZ9, a ratiometric fluorescent probe for mobile zinc. A plate reader-based assay using HcZ9 was developed, the accuracy of which is comparable to that of atomic absorption spectroscopy. We investigated zinc accumulation in prostatic cells and zinc levels in human seminal fluid. When normal and tumorigenic cells are bathed in zinc-enriched media, cellular mobile zinc is buffered and changes slightly, but total zinc levels increase significantly. Quantification of mobile and total zinc levels in human seminal plasma revealed that the two are positively correlated with a Pearson's coefficient of 0.73.