In this chapter, we describe the preparation of fluorescent quantum dots for imaging and measuring protein expression in cells. Quantum dots are nanocrystals that have numerous advantages for biomolecular detection compared with organic dyes and fluorescent proteins, but their large size has been a limiting factor. We describe the synthesis of nanoparticles smaller than 10 nm (smaller than an antibody), their attachment to monoclonal antibodies through click chemistry, characterization of the conjugates, and use for labeling of cellular antigens. We further discuss the unique advantages and challenges associated with this approach compared with conventional immunofluorescence techniques.
Keywords: Antibody; Cancer; Cytology; EGFR; Fluorescence; Imaging; Immunofluorescence; Microscopy; Nanocrystal; Nanotechnology; Protein; Single cell.