Combining two-photon excitation with fluorescence lifetime imaging

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 ima...

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Bibliographic Details
Published inIEEE engineering in medicine and biology magazine Vol. 18; no. 5; pp. 31 - 36
Main Authors Gerritsen, H.C., Vroom, J.M., de Grauw, C.J.
Format Journal Article
LanguageEnglish
Published United States IEEE 01.09.1999
Subjects
Aging
Bacteria - cytology
Biofilms
Biology
Confocal
Excitation
Fluoresceins - chemistry
Fluorescence
Hydrogen-Ion Concentration
Image recording
Imaging
Laser excitation
Laser modes
Lasers
Light scattering
Medical imaging
Microelectrodes
Microscopes
Microscopy
Microscopy, Confocal - instrumentation
Microscopy, Confocal - methods
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Optical filters
Optical imaging
Optical pumping
Optical resolving power
Probes
Rhodamines - chemistry
Space life sciences
Thermoplastic elastomers
Ultrafast optics
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Summary: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.
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ISSN:0739-5175
1937-4186
DOI:10.1109/51.790989