Instrumentation to rapidly acquire fluorescence wavelength-time matrices of biological tissues

Lloyd, William R.; Wilson, Robert H.; Chang, Ching Wei; Gillispie, Gregory D.; Mycek, Mary Ann

doi:10.1364/boe.1.000574

Cited by 17 publications

(7 citation statements)

References 27 publications

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“…However, their pulse-width is two orders of magnitude larger than modelocked fiber lasers. Semiconductor lasers also have been applied to fluorescence spectroscopy is 5,8,36 but their pJ pulse energy and limited wavelengths of operation no shorter to 375 nm make them unsuitable for fluorescence spectroscopy applications in collagen.…”

Section: B Data Acquisition Speedmentioning

confidence: 99%

“…Thus, for a more effective use of fluorescence lifetime(s) information, measurements of spectrally resolved fluorescence decay characteristics are required. For tissue diagnosis, this has typically been achieved through the use of either a scanning monochromator 1,8 or a set of band-pass filters 3,9 that can resolve the sample emission spectrum sequentially before each spectral intensity is time-resolved. The common methods for resolving the emission decay characteristics include time-correlated single photon counting (TCSPC), 10,11 time-gated, 12,13 and pulse sampling.…”

Section: Introductionmentioning

confidence: 99%

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

Yankelevich

Liu

et al. 2014

Review of Scientific Instruments

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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 Laguerreexpansion 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. © 2014 AIP Publishing LLC. [http://dx

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Section: B Data Acquisition Speedmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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

Yankelevich

Liu

et al. 2014

Review of Scientific Instruments

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“…FLIM has also been employed to image NADH distribution in live progenitor stem cells (Wright et al, ). When FLIM images are acquired at multiple wavelengths—i.e., multispectral FLIM (Becker et al, ; Owen et al, ; Lloyd et al, )—functional information from multiple fluorophores can be analyzed simultaneously. With this technique, NAD(P)H and FAD, two biologically relevant molecules related to cellular metabolism, can be preferentially detected with emission less than ∼500 nm for NAD(P)H and greater than ∼500 nm for FAD (Skala et al, ).…”

Section: Fluorescence Lifetime Imaging Microscopymentioning

confidence: 99%

FRAP, FLIM, and FRET: Detection and analysis of cellular dynamics on a molecular scale using fluorescence microscopy

Santos

Chang

Mycek

et al. 2015

Molecular Reproduction Devel

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The combination of fluorescent-probe technology plus modern optical microscopes allows investigators to monitor dynamic events in living cells with exquisite temporal and spatial resolution. Fluorescence recovery after photobleaching (FRAP), for example, has long been used to monitor molecular dynamics both within cells and on cellular surfaces. Although bound by the diffraction limit imposed on all optical microscopes, the combination of digital cameras and the application of fluorescence intensity information on large-pixel arrays have allowed such dynamic information to be monitored and quantified. Fluorescence lifetime imaging microscopy (FLIM), on the other hand, utilizes the information from an ensemble of fluorophores to probe changes in the local environment. Using either fluorescence-intensity or lifetime approaches, fluorescence resonance energy transfer (FRET) microscopy provides information about molecular interactions, with Ångstrom resolution. In this review, we summarize the theoretical framework underlying these methods and illustrate their utility in addressing important problems in reproductive and developmental systems.

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“…Wavelength-resolved curves are user-input within the fixed scanning window to 300 to 800 nm and resolution of ≥ 0.01 nm. Previously determined system signal-to-noise ratio was determined to be 100 (with 25 or greater laser pulses averaged per waveform, fluorophore concentration ≥ 1 µM [2]). …”

Section: Instrumentation -Clinically-compatible Specialized Transientmentioning

confidence: 99%

Fluorescence wavelength-time matrix acquisition for biomedical tissue diagnostics

et al. 2011

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A specialized transient digitizer system was developed for spectroscopic collection of fluorescence wavelength-time matrices (WTMs) from biological tissues. The system is compact, utilizes fiber optic probes for clinical compatibility, and offers rapid collection of high signal-to-noise ratio (>100) time-and wavelength-resolved fluorescence. The system is compatible with excitation sources operating in excess of 25 kHz. Wavelength-resolved measurement range is 300-800 nm with ≥ 0.01 nm steps. Time-resolved measurement depth is 128 ns with fixed 0.2 ns steps. The information-rich WTM data provides comprehensive fluorescence sensing capabilities, as demonstrated on tissue simulating phantoms. Extracting wavelength-resolved fluorophore lifetimes illustrates the potential of using the technology to resolve exogenous or endogenous fluorophore contributions in tissue samples in a clinical setting for tissue diagnostics and monitoring.

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Instrumentation to rapidly acquire fluorescence wavelength-time matrices of biological tissues

Cited by 17 publications

References 27 publications

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

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

FRAP, FLIM, and FRET: Detection and analysis of cellular dynamics on a molecular scale using fluorescence microscopy

Fluorescence wavelength-time matrix acquisition for biomedical tissue diagnostics

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