Oxygen microscopy by two-photon-excited phosphorescence.
High-resolution images of oxygen distributions in microheterogeneous samples are obtained by two-photon laser scanning microscopy (2P LSM), using a newly developed dendritic nanoprobe with internally enhanced two-photon absorption (2PA) cross-section. In this probe, energy is harvested by a 2PA antenna, which passes excitation onto a phosphorescent metalloporphyrin via intramolecular energy transfer. The 2P LSM allows sectioning of oxygen gradients with near diffraction-limited resolution, and lifetime-based acquisition eliminates dependence on the local probe concentration. The technique is validated on objects with a priori known oxygen distributions and applied to imaging of pO(2) in cells.
Finikova
Olga S
OS
Department of Biochemistry and Biophysics, University of Pennsylvannia, Philadelphia, PA 19104-6059, USA.
Lebedev
Artem Y
AY
Aprelev
Alexey
A
Troxler
Thomas
T
Gao
Feng
F
Garnacho
Carmen
C
Muro
Silvia
S
Hochstrasser
Robin M
RM
Vinogradov
Sergei A
SA
D002478:Cells, Cultured; D042783:Endothelial Cells; D006801:Humans; D008163:Luminescent Measurements; D008853:Microscopy / Q000379:methods*; D015394:Molecular Structure; D049329:Nanostructures; D010100:Oxygen / Q000737:chemistry*; D010777:Photochemistry; D017785:Photons
D010100:Oxygen