Second harmonic generating (SHG) nanoprobes for in vivo imaging.
Fluorescence microscopy has profoundly changed cell and molecular biology studies by permitting tagged gene products to be followed as they function and interact. The ability of a fluorescent dye to absorb and emit light of different wavelengths allows it to generate startling contrast that, in the best cases, can permit single molecule detection and tracking. However, in many experimental settings, fluorescent probes fall short of their potential due to dye bleaching, dye signal saturation, and tissue autofluorescence. Here, we demonstrate that second harmonic generating (SHG) nanoprobes can be used for in vivo imaging, circumventing many of the limitations of classical fluorescence probes. Under intense illumination, such as at the focus of a laser-scanning microscope, these SHG nanocrystals convert two photons into one photon of half the wavelength; thus, when imaged by conventional two-photon microscopy, SHG nanoprobes appear to generate a signal with an inverse Stokes shift like a fluorescent dye, but with a narrower emission. Unlike commonly used fluorescent probes, SHG nanoprobes neither bleach nor blink, and the signal they generate does not saturate with increasing illumination intensity. The resulting contrast and detectability of SHG nanoprobes provide unique advantages for molecular imaging of living cells and tissues.
Pantazis
Periklis
P
Division of Biology, California Institute of Technology, Beckman Institute 139-74, Pasadena, CA 91125, USA. pantazis@caltech.edu
Maloney
James
J
Wu
David
D
Fraser
Scott E
SE
D000818:Animals; D017609:Barium Compounds / Q000737:chemistry; D004625:Embryo, Nonmammalian / Q000737:chemistry / Q000166:cytology / Q000196:embryology; D007150:Immunohistochemistry; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D015335:Molecular Probes / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry / Q000648:ultrastructure; D036103:Nanotechnology / Q000379:methods*; D014025:Titanium / Q000737:chemistry; D015027:Zebrafish / Q000196:embryology / Q000378:metabolism
D017609:Barium Compounds; D015335:Molecular Probes; C024547:barium titanate(IV); D014025:Titanium