Lifetime-based tomographic multiplexing
Near-infrared (NIR) fluorescence tomography of multiple fluorophores has previously been limited by the bandwidth of the NIR spectral regime and the broad emission spectra of most NIR fluorophores. We describe in vivo tomography of three spectrally overlapping fluorophores using fluorescence lifetim...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | en_US |
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SPIE
2011
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| Online Access: | http://hdl.handle.net/1721.1/60943 |
| _version_ | 1811086512194846720 |
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| author | Raymond, Scott Bruce Boas, David A. Kumar, Anand T. N. Bacskai, Brian J. |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Raymond, Scott Bruce Boas, David A. Kumar, Anand T. N. Bacskai, Brian J. |
| author_sort | Raymond, Scott Bruce |
| collection | MIT |
| description | Near-infrared (NIR) fluorescence tomography of multiple fluorophores has previously been limited by the bandwidth of the NIR spectral regime and the broad emission spectra of most NIR fluorophores. We describe in vivo tomography of three spectrally overlapping fluorophores using fluorescence lifetime-based separation. Time-domain images are acquired using a voltage-gated, intensified charge-coupled device (CCD) in free-space transmission geometry with 750 nm Ti:sapphire laser excitation. Lifetime components are fit from the asymptotic portion of fluorescence decay curve and reconstructed separately with a lifetime-adjusted forward model. We use this system to test the in vivo lifetime multiplexing suitability of commercially available fluorophores, and demonstrate lifetime multiplexing in solution mixtures and in nude mice. All of the fluorophores tested exhibit nearly monoexponential decays, with narrow in vivo lifetime distributions suitable for lifetime multiplexing. Quantitative separation of two fluorophores with lifetimes of 1.1 and 1.37 ns is demonstrated for relative concentrations of 1:5. Finally, we demonstrate tomographic imaging of two and three fluorophores in nude mice with fluorophores that localize to distinct organ systems. This technique should be widely applicable to imaging multiple NIR fluorophores in 3-D. |
| first_indexed | 2024-09-23T13:27:07Z |
| format | Article |
| id | mit-1721.1/60943 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:27:07Z |
| publishDate | 2011 |
| publisher | SPIE |
| record_format | dspace |
| spelling | mit-1721.1/609432022-10-01T15:23:07Z Lifetime-based tomographic multiplexing Raymond, Scott Bruce Boas, David A. Kumar, Anand T. N. Bacskai, Brian J. Harvard University--MIT Division of Health Sciences and Technology Raymond, Scott Bruce Raymond, Scott Bruce Near-infrared (NIR) fluorescence tomography of multiple fluorophores has previously been limited by the bandwidth of the NIR spectral regime and the broad emission spectra of most NIR fluorophores. We describe in vivo tomography of three spectrally overlapping fluorophores using fluorescence lifetime-based separation. Time-domain images are acquired using a voltage-gated, intensified charge-coupled device (CCD) in free-space transmission geometry with 750 nm Ti:sapphire laser excitation. Lifetime components are fit from the asymptotic portion of fluorescence decay curve and reconstructed separately with a lifetime-adjusted forward model. We use this system to test the in vivo lifetime multiplexing suitability of commercially available fluorophores, and demonstrate lifetime multiplexing in solution mixtures and in nude mice. All of the fluorophores tested exhibit nearly monoexponential decays, with narrow in vivo lifetime distributions suitable for lifetime multiplexing. Quantitative separation of two fluorophores with lifetimes of 1.1 and 1.37 ns is demonstrated for relative concentrations of 1:5. Finally, we demonstrate tomographic imaging of two and three fluorophores in nude mice with fluorophores that localize to distinct organ systems. This technique should be widely applicable to imaging multiple NIR fluorophores in 3-D. National Institutes of Health (U.S) (EB000768) National Institutes of Health (U.S) (AG026240) National Institutes of Health (U.S) (T32 EB001680) 2011-02-14T19:54:56Z 2011-02-14T19:54:56Z 2010-08 2010-03 Article http://purl.org/eprint/type/JournalArticle 1083-3668 1560-2281 http://hdl.handle.net/1721.1/60943 Scott B. Raymond, David A. Boas, Brian J. Bacskai and Anand T. N. Kumar, "Lifetime-based tomographic multiplexing", J. Biomed. Opt. 15, 046011 (Aug 06, 2010); doi:10.1117/1.3469797 © 2010 Society of Photo-Optical Instrumentation Engineers en_US http://dx.doi.org/10.1117/1.3469797 Journal of Biomedical Optics Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf SPIE SPIE |
| spellingShingle | Raymond, Scott Bruce Boas, David A. Kumar, Anand T. N. Bacskai, Brian J. Lifetime-based tomographic multiplexing |
| title | Lifetime-based tomographic multiplexing |
| title_full | Lifetime-based tomographic multiplexing |
| title_fullStr | Lifetime-based tomographic multiplexing |
| title_full_unstemmed | Lifetime-based tomographic multiplexing |
| title_short | Lifetime-based tomographic multiplexing |
| title_sort | lifetime based tomographic multiplexing |
| url | http://hdl.handle.net/1721.1/60943 |
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