Multidimensional Fluorescence Imaging Applied to Biological Tissue

Elson, Daniel S.; Galletly, Neil; Talbot, Clifford; Requejo-Isidro, José; McGinty, James; Dunsby, Christopher; Lanigan, Peter M. P.; Munro, Ian; Benninger, Richard K.P.; Beule, Pieter De; Auksorius, Eigidijus; Hegyi, László; Sandison, Ann; Wallace, Andrew L.; Soutter, Pat; Neil, Mark A. A.; Lever, J. W.; Stamp, Gordon; French, P. M. W.

doi:10.1007/0-387-33016-x_22

Cited by 18 publications

(36 citation statements)

References 119 publications

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“…Possibly, more rigorous data analysis or a time-resolved fluorescence detection could be used to separate the overlapping fluorescences. Interestingly, Elson et al 29 reported differences in fluorescence lifetime images between healthy and diseased human articular cartilage.…”

Section: Discussionmentioning

confidence: 99%

Nondestructive fluorescence-based quantification of threose-induced collagen cross-linking in bovine articular cartilage

et al. 2012

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Abstract. Extensive collagen cross-linking affects the mechanical competence of articular cartilage: it can make the cartilage stiffer and more brittle. The concentrations of the best known cross-links, pyridinoline and pentosidine, can be accurately determined by destructive high-performance liquid chromatography (HPLC). We explore a nondestructive evaluation of cross-linking by using the intrinsic fluorescence of the intact cartilage. Articular cartilage samples from bovine knee joints were incubated in threose solution for 40 and 100 h to increase the collagen crosslinking. Control samples without threose were also prepared. Excitation-emission matrices at wavelengths of 220 to 950 nm were acquired from the samples, and the pentosidine and pyridinoline cross-links and the collagen concentrations were determined using HPLC. After the threose treatment, pentosidine and lysyl pyridinole (LP) concentrations increased. The intrinsic fluorescence, excited below 350 nm, decreased and was related to pentosidine [r ¼ −0.90, 240∕325 nm (excitation/emission)] or LP (r ¼ −0.85, 235∕285 nm) concentrations. Due to overlapping, the changes in emission could not be linked specifically to the recorded cross-links. However, the fluorescence signal enabled a nondestructive optical estimate of changes in the pentosidine and LP cross-linking of intact articular cartilage.

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Section: Discussionmentioning

confidence: 99%

Nondestructive fluorescence-based quantification of threose-induced collagen cross-linking in bovine articular cartilage

et al. 2012

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“…FLIM has been shown to provide label-free contrast between normal and diseased tissue, e.g. [8,[12][13][14], and is widely used to read out fluorescence sensors [15] and Fö rster resonant energy transfer (FRET) to probe molecular interactions [16]. We have previously demonstrated wide-field timegated FLIM endoscopy at low magnification [12] and here we report a confocal FLIM endomicroscope providing optically sectioned sub-cellular resolution in a clinically applicable instrument, which we have applied to imaging biological cells and tissue.…”

Section: Biophotonicsmentioning

confidence: 99%

A fluorescence lifetime imaging scanning confocal endomicroscope

Kennedy

Manning

Elson

et al. 2009

Journal of Biophotonics

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“…Optical methods based on fluorescence lifetime contrast such as time-resolved fluorescence spectroscopy (TRFS) and fluorescence lifetime imaging microscopy (FLIM) can detect biochemical and physiological transformations in tissue 5 and are known to improve the specificity of fluorescence measurements. [6][7][8] Several endogenous biomolecules, including structural proteins (e.g., collagen and elastin) and enzyme cofactors (e.g., NADH and FAD), are responsible for tissue autofluorescence. 9 Changes in the fluorescence properties of tissue are reflective of native fluorophore deposition and, thus, the relative contribution and distribution of individual fluorophores within the tissue.…”

Section: Introductionmentioning

confidence: 99%

Noninvasive Multimodal Evaluation of Bioengineered Cartilage Constructs Combining Time-Resolved Fluorescence and Ultrasound Imaging

Fite

Decaris

Sun

et al. 2011

Tissue Engineering Part C: Methods

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A multimodal diagnostic system that integrates time-resolved fluorescence spectroscopy, fluorescence lifetime imaging microscopy, and ultrasound backscatter microscopy is evaluated here as a potential tool for assessing changes in engineered tissue composition and microstructure nondestructively and noninvasively. The development of techniques capable of monitoring the quality of engineered tissue, determined by extracellular matrix (ECM) content, before implantation would alleviate the need for destructive assays over multiple time points and advance the widespread development and clinical application of engineered tissues. Using a prototype system combining time-resolved fluorescence spectroscopy, FLIM, and UBM, we measured changes in ECM content occurring during chondrogenic differentiation of equine adipose stem cells on 3D biodegradable matrices. The optical and ultrasound results were validated against those acquired via conventional techniques, including collagen II immunohistochemistry, picrosirius red staining, and measurement of construct stiffness. Current results confirm the ability of this multimodal approach to follow the progression of tissue maturation along the chondrogenic lineage by monitoring ECM production (namely, collagen type II) and by detecting resulting changes in mechanical properties of tissue constructs. Although this study was directed toward monitoring chondrogenic tissue maturation, these data demonstrate the feasibility of this approach for multiple applications toward engineering other tissues, including bone and vascular grafts.

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Multidimensional Fluorescence Imaging Applied to Biological Tissue

Cited by 18 publications

References 119 publications

Nondestructive fluorescence-based quantification of threose-induced collagen cross-linking in bovine articular cartilage

Nondestructive fluorescence-based quantification of threose-induced collagen cross-linking in bovine articular cartilage

A fluorescence lifetime imaging scanning confocal endomicroscope

Noninvasive Multimodal Evaluation of Bioengineered Cartilage Constructs Combining Time-Resolved Fluorescence and Ultrasound Imaging

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