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Characterization of Leadmium Green for Fluorescence Detection of Intracellular Cadmium
Author(s) -
Malaiyandi Latha M.,
Sharthiya Harsh,
Dineley Kirk E.
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.824.2
Subject(s) - cadmium , zinc , chemistry , fluorescence , intracellular , ionophore , biophysics , biochemistry , membrane , biology , physics , organic chemistry , quantum mechanics
Cadmium is a common industrial pollutant, but the study of cadmium cytotoxicity is difficult in part due to the absence of a probe that can detect cadmium in living cells without the confounds of contaminating metals. Leadmium Green (LG) is a commercially available, small molecule, fluorescent probe advertised as a detector of intracellular cadmium and lead. LG has been used to monitor cadmium sequestration in plant cells, heavy metal export in a protozoan, and lead absorption by vascular endothelial cells. However very little information is available regarding its affinity and selectivity for cadmium, lead, and other metals. Using a cell‐impermeant form, we evaluated the in vitro selectivity of LG using spectrofluorometry. Consistent with manufacturer's claims, LG responded robustly to cadmium (K D 603nM) and also lead (K D 8.65nM) in a concentration‐dependent manner, and furthermore proved insensitive to calcium, cobalt, manganese and nickel. However, LG also responded to zinc with a K D of 81.9nM. Using fluorescence microscopy, we evaluated the utility of LG in live mouse hippocampal HT22 cells. We demonstrated that LG detects ionophore‐mediated elevations of cadmium or zinc in a concentration‐dependent manner. In comparison to zinc changes reported by the widely used zinc fluorescent probe FluoZin‐3, LG yielded a relatively small dynamic range. In conclusion, LG may have utility as an intracellular cadmium probe, however data must be interpreted cautiously due to potential contribution of zinc to the signal.