Design and evaluation of a device for fast multispectral time-resolved fluorescence spectroscopy and imaging.

The application of time-resolved fluorescence spectroscopy (TRFS) to in vivo tissue diagnosis requires a method for fast acquisition of fluorescence decay profiles in multiple spectral bands. This study focusses on development of a clinically compatible fiber-optic based multispectral TRFS (ms-TRFS) system together with validation of its accuracy and precision for fluorescence lifetime measurements. It also presents the expansion of this technique into an imaging spectroscopy method. A tandem array of dichroic beamsplitters and filters was used to record TRFS decay profiles at four distinct spectral bands where biological tissue typically presents fluorescence emission maxima, namely, 390, 452, 542, and 629 nm. Each emission channel was temporally separated by using transmission delays through 200 μm diameter multimode optical fibers of 1, 10, 19, and 28 m lengths. A Laguerre-expansion deconvolution algorithm was used to compensate for modal dispersion inherent to large diameter optical fibers and the finite bandwidth of detectors and digitizers. The system was found to be highly efficient and fast requiring a few nano-Joule of laser pulse energy and <1 ms per point measurement, respectively, for the detection of tissue autofluorescent components. Organic and biological chromophores with lifetimes that spanned a 0.8-7 ns range were used for system validation, and the measured lifetimes from the organic fluorophores deviated by less than 10% from values reported in the literature. Multi-spectral lifetime images of organic dye solutions contained in glass capillary tubes were recorded by raster scanning the single fiber probe in a 2D plane to validate the system as an imaging tool. The lifetime measurement variability was measured indicating that the system provides reproducible results with a standard deviation smaller than 50 ps. The ms-TRFS is a compact apparatus that makes possible the fast, accurate, and precise multispectral time-resolved fluorescence lifetime measurements of low quantum efficiency sub-nanosecond fluorophores.

[1]  Jin-Ho Choy,et al.  Photophysical Properties of Hemicyanine Dyes Intercalated in Na−Fluorine Mica , 2000 .

[2]  P. French,et al.  Multifocal multiphoton excitation and time correlated single photon counting detection for 3-D fluorescence lifetime imaging. , 2007, Optics express.

[3]  Paritosh Pande,et al.  Fully automated deconvolution method for on-line analysis of time-resolved fluorescence spectroscopy data based on an iterative Laguerre expansion technique. , 2009, Journal of biomedical optics.

[4]  William R. Lloyd,et al.  Instrumentation to rapidly acquire fluorescence wavelength-time matrices of biological tissues , 2010, Biomedical optics express.

[5]  R. Steiner,et al.  SLIM: A new method for molecular imaging , 2007, Microscopy research and technique.

[6]  Manoj Kumbhakar,et al.  Photophysical properties of coumarin-120: Unusual behavior in nonpolar solvents , 2003 .

[7]  D. Elson,et al.  Fluorescence lifetime imaging microscopy for brain tumor image-guided surgery. , 2010, Journal of biomedical optics.

[8]  E. Fujimori.  Ultraviolet light- and ozone-induced changes in pyridinoline, a trisubstituted 3-hydroxypyridinium crosslink of collagen. , 1985, Biochimica et biophysica acta.

[9]  Qiyin Fang,et al.  Intraoperative delineation of primary brain tumors using time-resolved fluorescence spectroscopy. , 2010, Journal of biomedical optics.

[10]  Laura Marcu,et al.  Fluorescence lifetime imaging for the characterization of the biochemical composition of atherosclerotic plaques. , 2011, Journal of biomedical optics.

[11]  D. Neckers,et al.  TYPE I AND TYPE II SENSITIZERS BASED ON ROSE BENGAL ONIUM SALTS * , 1988, Photochemistry and photobiology.

[12]  Guilford Jones,et al.  Solvent effects on emission yield and lifetime for coumarin laser dyes. Requirements for a rotatory decay mechanism , 1983 .

[13]  W S Grundfest,et al.  Discrimination of Human Coronary Artery Atherosclerotic Lipid-Rich Lesions by Time-Resolved Laser-Induced Fluorescence Spectroscopy , 2001, Arteriosclerosis, thrombosis, and vascular biology.

[14]  P. French,et al.  Wide-field fluorescence lifetime imaging of cancer , 2010, Biomedical optics express.

[15]  Laura Marcu,et al.  Dynamic tissue analysis using time- and wavelength-resolved fluorescence spectroscopy for atherosclerosis diagnosis , 2011, Optics express.

[16]  D. Schweitzer,et al.  Towards metabolic mapping of the human retina , 2007, Microscopy research and technique.

[17]  Mark A A Neil,et al.  Fluorescence lifetime imaging by using time-gated data acquisition. , 2007, Applied optics.

[18]  G. Keiser.  Optical Fiber Communications , 1983 .

[19]  J. Lakowicz.  Principles of fluorescence spectroscopy , 1983 .

[20]  Laura Marcu,et al.  Simultaneous time- and wavelength-resolved fluorescence spectroscopy for near real-time tissue diagnosis. , 2008, Optics letters.

[21]  Laura Marcu,et al.  Multispectral fluorescence lifetime imaging system for intravascular diagnostics with ultrasound guidance: in vivo validation in swine arteries , 2014, Journal of biophotonics.

[22]  Kenneth G. Spears,et al.  Hydrogen bond strengths from solvent-dependent lifetimes of Rose Bengal dye , 1978 .

[23]  Cristina Kurachi,et al.  Noninvasive evaluation of oral lesions using depth‐sensitive optical spectroscopy , 2009, Cancer.

[24]  J. M. Morris,et al.  Picosecond Fluorescence Studies of Xanthene Dyes , 1977 .

[25]  Shuna Cheng,et al.  Flexible endoscope for continuous in vivo multispectral fluorescence lifetime imaging. , 2013, Optics letters.

[26]  A. Fletcher,et al.  Fluorescence quantum yields of some rhodamine dyes , 1982 .

[27]  T Joshua Pfefer,et al.  Temporally and spectrally resolved fluorescence spectroscopy for the detection of high grade dysplasia in Barrett's esophagus , 2003, Lasers in surgery and medicine.

[28]  Laura Marcu,et al.  Fluorescence Lifetime Techniques in Medical Applications , 2012, Annals of Biomedical Engineering.

[29]  D. Schweitzer,et al.  In vivo measurement of time-resolved autofluorescence at the human fundus. , 2004, Journal of biomedical optics.

[30]  M. Mycek,et al.  Handbook of Biomedical Fluorescence , 2003 .

[31]  W S Grundfest,et al.  Time-resolved Fluorescence Spectra of Arterial Fluorescent Compounds: Reconstruction with the Laguerre Expansion Technique , 2000, Photochemistry and photobiology.

[32]  E. Bakienė,et al.  Characterization of biological materials by frequency-domain fluorescence lifetime measurements using ultraviolet light-emitting diodes , 2008 .

[33]  Laura Marcu,et al.  Design, construction, and validation of a rotary multifunctional intravascular diagnostic catheter combining multispectral fluorescence lifetime imaging and intravascular ultrasound , 2012, Journal of biomedical optics.

[34]  D Fujimoto,et al.  Isolation and characterization of a fluorescent material in bovine achilles tendon collagen , 1977 .

[35]  J. Penn,et al.  Evaluation of single-photon-counting measurements of excited-state lifetimes. , 1982, Proceedings of the National Academy of Sciences of the United States of America.