Low-frequency wide-field fluorescence lifetime imaging using a high-power near-infrared light-emitting diode light source.
Fluorescence lifetime imaging (FLi) could potentially improve exogenous near-infrared (NIR) fluorescence imaging, because it offers the capability of discriminating a signal of interest from background, provides real-time monitoring of a chemical environment, and permits the use of several different fluorescent dyes having the same emission wavelength. We present a high-power, LED-based, NIR light source for the clinical translation of wide-field (larger than 5 cm in diameter) FLi at frequencies up to 35 MHz. Lifetime imaging of indocyanine green (ICG), IRDye 800-CW, and 3,3(')-diethylthiatricarbocyanine iodide (DTTCI) was performed over a large field of view (10 cm by 7.5 cm) using the LED light source. For comparison, a laser diode light source was employed as a gold standard. Experiments were performed both on the bench by diluting the fluorescent dyes in various chemical environments in Eppendorf tubes, and in vivo by injecting the fluorescent dyes mixed in Matrigel subcutaneously into CD-1 mice. Last, measured fluorescence lifetimes obtained using the LED and the laser diode sources were compared with those obtained using a state-of-the-art time-domain imaging system and with those previously described in the literature. On average, lifetime values obtained using the LED and the laser diode light sources were consistent, exhibiting a mean difference of 3% from the expected values and a coefficient of variation of 12%. Taken together, our study offers an alternative to laser diodes for clinical translation of FLi and explores the use of relatively low frequency modulation for in vivo imaging.
Gioux
Sylvain
S
Boston University, 48 Cummington Street, Boston, Massachusetts 02215, USA.
Lomnes
Stephen J
SJ
Choi
Hak Soo
HS
Frangioni
John V
JV
D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000379:methods*; D007259:Infrared Rays; D008029:Lighting / Q000295:instrumentation*; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D015203:Reproducibility of Results; D012666:Semiconductors; D012680:Sensitivity and Specificity
Appl Opt. 1996 May 1;35(13):2304-14
21085367
Mol Imaging. 2007 Jul-Aug;6(4):237-46
17711779
Nano Lett. 2009 Jun;9(6):2354-9
19422261
Sens Actuators B Chem. 1995 Oct;29(1-3):16-24
33867678
Appl Opt. 1996 May 1;35(13):2325-32
21085369
Biophys J. 1995 Apr;68(4):1574-82
7787043
Appl Opt. 1994 Apr 1;33(10):1963-74
20885531
J Biomed Opt. 2008 Sep-Oct;13(5):054042
19021422
Proc Natl Acad Sci U S A. 1992 Feb 15;89(4):1271-5
1741380
J Biomed Opt. 2009 May-Jun;14(3):034045
19566337
Ann Surg Oncol. 2006 Dec;13(12):1671-81
17009138
Appl Opt. 1997 Apr 1;36(10):2260-72
18253202
Breast Cancer. 2005;12(3):211-5
16110291
Appl Opt. 1996 Jul 1;35(19):3746-58
21102772
Ann Surg Oncol. 2009 Oct;16(10):2943-52
19582506
Dig Surg. 2008;25(2):103-8
18379188
Opt Lett. 1996 Jan 15;21(2):158-60
19865337
J Plast Reconstr Aesthet Surg. 2009 Oct;62(10):e373-8
18556255
Appl Opt. 1989 Jun 15;28(12):2331-6
20555520
Annu Rev Biophys Bioeng. 1984;13:105-24
6378065
Ann Surg Oncol. 2008 Jun;15(6):1640-3
18379850
Biophys J. 2007 Oct 15;93(8):2892-9
17573433
Photochem Photobiol. 1999 Jul;70(1):87-94
10420847
Am J Physiol Heart Circ Physiol. 2007 Jun;292(6):H3109-18
17307997
J Biomed Opt. 2009 Nov-Dec;14(6):060504
20059235
IEEE Trans Med Imaging. 2008 Aug;27(8):1152-63
18672432
Appl Opt. 1997 Jan 1;36(1):116-24
18250652
Mol Imaging. 2009 May-Jun;8(3):156-65
19723473
Biotechnol Prog. 1997 Sep-Oct;13(5):669-80
9336987
J Microsc. 2006 May;222(Pt 2):135-40
16774522
Photochem Photobiol Sci. 2004 Aug;3(8):795-801
15295637
Nat Biotechnol. 2007 Oct;25(10):1165-70
17891134
J Biomed Opt. 2005 Sep-Oct;10(5):051403
16292940
Anal Biochem. 1992 May 1;202(2):316-30
1519759
Proc SPIE Int Soc Opt Eng. 2008;6848:684807
19655029
Photochem Photobiol. 1997 Aug;66(2):229-36
9277142
Appl Opt. 1999 Aug 1;38(22):4930-8
18323983
J Biomed Opt. 2009 Mar-Apr;14(2):024012
19405742
World J Surg. 2008 Sep;32(9):1924-9
18330628