Preparation of a copper-based fluorescent probe for nitric oxide and its use in mammalian cultured cells.
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.
Lim
Mi Hee
MH
Department of Chemistry, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA. miheelim@alum.mit.edu
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
D005456:Fluorescent Dyes; D009569:Nitric Oxide; D003300:Copper