Combining two-photon excitation with fluorescence lifetime imaging

Author

Gerritsen, H.C. ; Vroom, J.M. ; de Grauw, C.J.

Author_Institution

Debye Res. Inst., Utrecht Univ., Netherlands

Volume

18

Issue

5

fYear

1999

Firstpage

31

Lastpage

36

Abstract

Two-photon excitation (TPE) imaging with a microscope objective that is properly matched to the sample results in a two- to four-times larger penetration depth than with confocal laser scanning microscopy. The pulsed light source present in a TPE microscope makes time-gated fluorescence lifetime imaging an attractive alternative for quantitative imaging of ion concentrations. Using TPE fluorescence lifetime imaging, in-depth imaging of pH in biofilm can be accomplished.

Keywords

bio-optics; biological techniques; biomedical imaging; fluorescence; optical microscopy; pH measurement; radiative lifetimes; two-photon processes; dental biofilm; fluorescence lifetime imaging; improved penetration depth; in-depth imaging of pH; photobleaching extent; pulsed light source; quantitative imaging; resolution effects; time-gated imaging; two-photon excitation imaging; Aging; Fluorescence; Laser excitation; Laser modes; Microscopy; Optical filters; Optical imaging; Optical pumping; Probes; Ultrafast optics; Bacteria; Biofilms; Fluoresceins; Hydrogen-Ion Concentration; Microscopy, Confocal; Microscopy, Fluorescence; Rhodamines;

fLanguage

English

Journal_Title

Engineering in Medicine and Biology Magazine, IEEE

Publisher

ieee

ISSN

0739-5175

Type

jour

DOI

10.1109/51.790989

Filename

790989