title: Hybrid confocal Raman fluorescence microscopy on single cells using semiconductor quantum dots.
abstract: We have overcome the traditional incompatibility of Raman microscopy with fluorescence microscopy by exploiting the optical properties of semiconductor fluorescent quantum dots (QDs). Here we present a hybrid Raman fluorescence spectral imaging approach for single-cell microscopy applications. We show that resonant Raman imaging of flavocytochrome b558 at 413.1 nm excitation in QD-labeled neutrophilic granulocytes or nonresonant Raman imaging of proteins and lipids at 647.1 nm excitation in QD-labeled macrophages can be integrated with linear one-photon excitation and nonlinear continuous-wave two-photon excitation fluorescence microscopy of QDs, respectively. The enhanced information content of these two hybrid Raman fluorescence methods provides new multiplexing possibilities for single-cell optical microscopy and intracellular chemical analysis.
authors.affiliation: MESA+ Institute for Nanotechnology, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands. h.w.j.vanmanen@tnw.utwente.nl
authors.forename: Henk-Jan
authors.initials: HJ
authors.lastname: van Manen
authors.forename: Cees
authors.initials: C
authors.lastname: Otto
mesh_terms: D002478:Cells, Cultured; D006801:Humans; D007089:Image Enhancement / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D009504:Neutrophils / Q000166:cytology*; D045663:Quantum Dots; D012666:Semiconductors; D013059:Spectrum Analysis, Raman / Q000379:methods*