Hybrid confocal Raman fluorescence microscopy on single cells using semiconductor quantum dots.
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.
van Manen
Henk-Jan
HJ
MESA+ Institute for Nanotechnology, University of Twente, P.O. Box 217, 7500 AE Enschede, The Netherlands. h.w.j.vanmanen@tnw.utwente.nl
Otto
Cees
C
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*