Coppersensor 1

Coppersensor-1 (CS1) is a boron dipyrromethene (BODIPY)-based fluorescent sensor for selective and sensitive detection of copper(I) ions (Cu+) in biological samples, including live cells[1].

Coppersensor 1 (CS1) comprising a boron dipyrromethene (BODIPY) chromophore coupled to a thioether-rich receptor, has a picomolar affinity for Cu+ with high selectivity over competing cellular metal ions. CS1 fluorescence increases up to 10-fold on binding to Cu+[1]. Guidelines[1] (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).1. Coppersensor 1 (CS1) is dissolved in DMSO to obtain a 1 mM stock solution and further diluted before use. 2. cells are growing in medium, adding 10 mM CuCl2 stock solution to give a final copper concentration of 100 μM, and incubating for 1-8 h. 3. Adding competing chelator (N-ethyl-3,6,12,15-tetrathia-9-monoazaheptadecane; 100 mM) to copper-supplemented cells to give a final chelator concentration of 500 μM. Mix thoroughly, and incubate in the dark for 5 min at 25°C (A 5- to 10-fold excess of chelator over copper supplementation gives optimal results).4. Cells are incubated with 1 mM Coppersensor 1 solution to give a final dye concentration of 5 mM. Mix thoroughly. (Higher concentrations of dye may result in high levels of background fluorescence.) Incubating for 5-20 min in the dark at 25°C or 37°C.5. CS1 can be imaged using any type of fluorescence microscope, including epifluorescence, confocal and multiphoton. For standard confocal experiments, best results are obtained with 543 nm excitation to match the absorption maximum of the apo and Cu+-bound probe.

[1]. Evan W Miller, et al. Preparation and use of Coppersensor-1, a synthetic fluorophore for live-cell copper imaging. Nat Protoc. 2006;1(2):824-7.