A UV–Visible–NIR fluorescence lifetime imaging microscope for laser-based biological sensing with picosecond resolution
dc.contributor.author | Dragnev, K. H. | en_US |
dc.contributor.author | Zhong, W. | en_US |
dc.contributor.author | Dmitrovsky, E. | en_US |
dc.contributor.author | Beamish, J. A. | en_US |
dc.contributor.author | Mycek, Mary-Ann | en_US |
dc.contributor.author | Sloboda, R. D. | en_US |
dc.contributor.author | Urayama, P. | en_US |
dc.contributor.author | Minn, F. K. | en_US |
dc.date.accessioned | 2006-09-08T20:02:23Z | |
dc.date.available | 2006-09-08T20:02:23Z | |
dc.date.issued | 2003-05 | en_US |
dc.identifier.citation | Urayama, P.; Zhong, W.; Beamish, J.A.; Minn, F.K.; Sloboda, R.D.; Dragnev, K.H.; Dmitrovsky, E.; Mycek, M.-A.; (2003). "A UV–Visible–NIR fluorescence lifetime imaging microscope for laser-based biological sensing with picosecond resolution." Applied Physics B Lasers and Optics 76(5): 483-496. <http://hdl.handle.net/2027.42/42165> | en_US |
dc.identifier.issn | 0946-2171 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/42165 | |
dc.description.abstract | This article describes the design and characterization of a wide-field, time-domain fluorescence lifetime imaging microscopy (FLIM) system developed for picosecond time-resolved biological imaging. The system consists of a nitrogen-pumped dye laser for UV–visible–NIR excitation (337.1–960 nm), an epi-illuminated microscope with UV compatible optics, and a time-gated intensified CCD camera with an adjustable gate width (200 ps-10 -3 s) for temporally resolved, single-photon detection of fluorescence decays with 9.6-bit intensity resolution and 1.4-μm spatial resolution. Intensity measurements used for fluorescence decay calculations are reproducible to within 2%, achieved by synchronizing the ICCD gate delay to the excitation laser pulse via a constant fraction optical discriminator and picosecond delay card. A self-consistent FLIM system response model is presented, allowing for fluorescence lifetimes (0.6 ns) significantly smaller than the FLIM system response (1.14 ns) to be determined to 3% of independently determined values. The FLIM system was able to discriminate fluorescence lifetime differences of at least 50 ps. The spectral tunability and large temporal dynamic range of the system are demonstrated by imaging in living human cells: UV-excited endogenous fluorescence from metabolic cofactors (lifetime ∼1.4 ns); and 460-nm excited fluorescence from an exogenous oxygen-quenched ruthenium dye (lifetime ∼400 ns). | en_US |
dc.format.extent | 2397257 bytes | |
dc.format.extent | 3115 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Springer-Verlag | en_US |
dc.subject.other | Legacy | en_US |
dc.subject.other | PACS: 87.64.-T; 87.57.-S | en_US |
dc.title | A UV–Visible–NIR fluorescence lifetime imaging microscope for laser-based biological sensing with picosecond resolution | en_US |
dc.type | Article | en_US |
dc.subject.hlbsecondlevel | Physics | en_US |
dc.subject.hlbsecondlevel | Mathematics | en_US |
dc.subject.hlbtoplevel | Science | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Dept. of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA, US | en_US |
dc.contributor.affiliationum | Dept. of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA, US | en_US |
dc.contributor.affiliationother | Dept. of Physics and Astronomy, Dartmouth College, Hanover, NH 03755, USA, US | en_US |
dc.contributor.affiliationother | Dept. of Medicine, Dartmouth Medical School, Hanover, NH 03755, USA, US | en_US |
dc.contributor.affiliationother | Dept. of Physics and Astronomy, Dartmouth College, Hanover, NH 03755, USA, US | en_US |
dc.contributor.affiliationother | Dept. of Biological Science, Dartmouth College, Hanover, NH 03755, USA, US | en_US |
dc.contributor.affiliationother | Dept. of Physics and Astronomy, Dartmouth College, Hanover, NH 03755, USA, US | en_US |
dc.contributor.affiliationother | Dept. of Medicine, Dartmouth Medical School, Hanover, NH 03755, USA, US | en_US |
dc.contributor.affiliationumcampus | Ann Arbor | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/42165/1/30760483.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1007/s00340-003-1152-4 | en_US |
dc.identifier.source | Applied Physics B Lasers and Optics | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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