title: Preparation of a copper-based fluorescent probe for nitric oxide and its use in mammalian cultured cells.
abstract: A procedure for the preparation of a copper(II) complex (CuFL) as a fluorescent nitric oxide (NO) detector is described. The fluorescein-based ligand FL can be synthesized in seven reaction steps (overall yield approximately 20%), typically requiring a total time of 9 days. The CuFL probe allows for the detection of NO produced in mammalian cultured cells. The detailed protocol for the use of CuFL for imaging NO in human neuroblastoma SK-N-SH cells takes a total time of approximately 26 h. This includes plating cells on six-well tissue culture plates or imaging dishes, treatment with CuFL, stimulation of NO synthases and imaging by fluorescence microscopy.
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. miheelim@alum.mit.edu
authors.forename: Mi Hee
authors.initials: MH
authors.lastname: Lim
mesh_terms: D002478:Cells, Cultured; D003300:Copper / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry; D006801:Humans; D008856:Microscopy, Fluorescence; D015336:Molecular Probe Techniques; D015394:Molecular Structure; D009569:Nitric Oxide / Q000302:isolation & purification* / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D005456:Fluorescent Dyes; D009569:Nitric Oxide; D003300:Copper