A hyperspectral fluorescence lifetime probe for skin cancer diagnosis

Beule, Pieter De; Dunsby, Chris; Galletly, Neil; Stamp, Gordon W.; Chu, A.C.; Anand, Uma; Anand, Praveen; Benham, C.D.; Naylor, Alan; French, Paul M. W.

doi:10.1063/1.2818785

Cited by 89 publications

(72 citation statements)

References 38 publications

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“…The time-resolved spectrometer (B) has been described previously [42,45] and was configured as shown in Figure 2. Two pulsed laser sources were used for excitation: a frequency tripled ultrafast Yb : glass fibre laser (UVPower355, Fianium Ltd., UK), which provided 10 ps pulses at 355 nm with a repetition rate of 37.1 MHz; and a diode laser (LDH-P-C-440B, PicoQuant GmbH, Germany), which emitted 50-150 ps pulses at 445 nm with an adjustable repetition rate of up to 40 MHz.…”

Section: Time-resolved Fibre-optic Probe Spectrofluorometer (B)mentioning

confidence: 99%

“…Instruments developed to monitor both fluorescence lifetime and spectra of tissue include the work of Glanzmann et al [40], Pitts et al [41], de Beule et al [42] and Sun et al [43]. There are also systems capable of measuring other optical properties, such as diffuse reflectance, in addition to the fluorescence, e.g.…”

Section: Introductionmentioning

confidence: 99%

“…There have been several recent examples of the application of such instruments to skin including previous work by our group on ex vivo tissue that showed significant differences between the fluorescence lifetimes of healthy and diseased tissue in non-melanoma skin cancer [24,42]. In addition, Rajaram et al studied non-melanoma skin cancer in vivo using a combination of diffuse reflectance and fluorescence spectroscopy [33], while Blackwell et al used an instrument recording fluorescence decays in four spectral channels to compare the fluorescence signature of human skin from diabetic and non-diabetic test subjects in vivo [44].…”

Section: Introductionmentioning

confidence: 99%

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In vivo measurements of diffuse reflectance and time‐resolved autofluorescence emission spectra of basal cell carcinomas

Thompson

Coda

Sørensen

et al. 2012

Journal of Biophotonics

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Journal of BIOPHOTONICSWe present a clinical investigation of diffuse reflectance and time-resolved autofluorescence spectra of skin cancer with an emphasis on basal cell carcinoma. A total of 25 patients were measured using a compact steady-state diffuse reflectance/fluorescence spectrometer and a fibre-optic-coupled multispectral time-resolved spectrofluorometer. Measurements were performed in vivo prior to surgical excision of the investigated region. Singular value decomposition was used to reduce the dimensionality of steady state diffuse reflectance and fluorescence spectra. Linear discriminant analysis was then applied to the measurements of basal cell carcinomas (BCCs) and used to predict the tissue disease state with a leave-one-out methodology. This approach was able to correctly diagnose 87% of the BCCs. With 445 nm excitation a decrease in the spectrally averaged fluorescence lifetime was observed between normal tissue and BCC lesions with a mean value of 886 ps. Furthermore, the fluorescence lifetime for BCCs was lower than that of the surrounding healthy tissue in all cases and statistical analysis of the data revealed that this decrease was significant (p ¼ 0.002).Schematic diagrams of the two spectrometers showing the steady state spectrometer measurement head (top) and the optical layout of the time-resolved system (bottom).

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Section: Time-resolved Fibre-optic Probe Spectrofluorometer (B)mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In vivo measurements of diffuse reflectance and time‐resolved autofluorescence emission spectra of basal cell carcinomas

Thompson

Coda

Sørensen

et al. 2012

Journal of Biophotonics

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“…Obtaining reliable results from biological tissues with MSP approach is then greatly simplified and becomes accessible to nonexperts without using the complex and time consuming fitting procedure generally associated with SLIM image analysis. It is thus conceivable to apply our MSP approach to facilitate tumor detections for instance, which are performed until now with simultaneous time-and wavelength-resolved spectroscopy due to the complexity of the data analysis (28,29). Thanks to the spectrally and temporally resolved visual segmentation of our MSP approach, we might indeed envisage to realize a fast and visual diagnostic of cancerous tumor imaging without risk of error.…”

Section: Discussionmentioning

confidence: 99%

Three‐dimensional polar representation for multispectral fluorescence lifetime imaging microscopy

et al. 2009

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Multispectral fluorescence lifetime imaging microscopy is a promising and powerful technique for discriminating multiply labeled samples and for detecting molecular interactions inside thick, heterogeneous, and light-scattering milieu such as tissue. The fast and correct analysis of the spectral and lifetime images constitutes a major challenge, which requires a high level of expertise. We present here a new approach that considerably simplifies this analysis avoiding complex fitting algorithm strategies and permitting a fast and visual graphical representation of the fluorescence lifetimes. By transforming the experimental data from time domain to frequency domain for each spectral channel, we calculate the multispectral polar representation and demonstrate its interest on multiply fluorescent labeled sample. We further apply it on Förster resonance energy transfer (FRET) experiments and demonstrate that FRET measurements with a high level of precision can be performed. With addition of emission wavelength as third dimension in the polar representation, autofluorescence emitted by the sample is thus clearly identified. Analysis artifacts induced by the sample or by fitting algorithm choice become then totally inexistent. ' 2009 International Society for Advancement of Cytometry Key terms multispectral FLIM; SLIM; biological tissue; molecular interactions; FRET; phasor IN addition to intensity and wavelength, lifetime is a supplementary source of contrast in fluorescence microscopy, which greatly increases access to the biological information in living cells. Fluorescence Lifetime Imaging Microscopy (FLIM) has indeed been successfully applied to differentiate spectrally undistinguishable molecular species (1) and to map local modifications in labeled samples in terms of ion concentration (2), pH (3), and oxygen (4). FLIM has also been widely used to explore conformational change of proteins (5) or to separate interacting and non-interacting molecular fractions in Förster Resonance Energy Transfer (FRET) experiments (6,7).However, because of the low number of collected photons per pixel and the variability of the molecular environment inside a living cell, FLIM is usually limited to two lifetime components in the same pixel. This restriction makes it difficult to extract biologically relevant information from autofluorescent, heterogeneous, lightscattering samples such as biological tissue. Because of optical properties of tissue, a dedicated temporally and spectrally resolved system (called SLIM for Spectral and Lifetime Imaging Microscopy) is necessary for example to be able to explore molecular interactions on a nanometric scale (with FRET experiments).Temporal and spectral measurements can be performed using two approaches, either frequency domain (8,9) or time domain (10-12) measurements. In both cases, the determination of lifetimes and contributions of each molecular species are mainly achieved by fitting the collected data at each pixel using equations with multiple

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“…It is to be expected that the exterior surface, e.g. skin cancer (38), the gastrointestinal tract, e.g. cancer of the mouth, esophagus, stomach, colon, and the outer surfaces of organs in the abdomen, e.g.…”

Section: Current (Pre)clinical Applicationmentioning

confidence: 99%

Advances in cellular, subcellular, and nanoscale imaging in vitro and in vivo

Wessels¹,

Yamauchi

Hoffman

et al. 2010

Cytometry Pt A

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This review focuses on technical advances in fluorescence microscopy techniques including laser scanning techniques, fluorescence-resonance energy transfer (FRET) microscopy, fluorescence lifetime imaging (FLIM), stimulated emission depletion (STED)-based super-resolution microscopy, scanning confocal endomicroscopes, thinsheet laser imaging microscopy (TSLIM), and tomographic techniques such as early photon tomography (EPT) as well as on clinical laser-based endoscopic and microscopic techniques. We will also discuss the new developments in the field of fluorescent dyes and fluorescent genetic reporters that enable new possibilities in high-resolution and molecular imaging both in in vitro and in vivo. Small animal and tissue imaging benefit from the development of new fluorescent proteins, dyes, and sensing constructs that operate in the far red and near-infrared spectrum. ' 2010 International Society for Advancement of Cytometry

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A hyperspectral fluorescence lifetime probe for skin cancer diagnosis

Cited by 89 publications

References 38 publications

In vivo measurements of diffuse reflectance and time‐resolved autofluorescence emission spectra of basal cell carcinomas

In vivo measurements of diffuse reflectance and time‐resolved autofluorescence emission spectra of basal cell carcinomas

Three‐dimensional polar representation for multispectral fluorescence lifetime imaging microscopy

Advances in cellular, subcellular, and nanoscale imaging in vitro and in vivo

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