Publications
550 results found
Xavier GDS, Mondragon A, Mitchell R, et al., 2013, Defective glucose homeostasis in mice inactivated selectively for <i>Tcf7l2</i> in the adult beta cell with an Ins1-controlled Cre, 49th Annual Meeting of the European-Association-for-the-Study-of-Diabetes (EASD), Publisher: SPRINGER, Pages: S142-S142, ISSN: 0012-186X
Warren SC, Margineanu A, Alibhai D, et al., 2013, Rapid Global Fitting of Large Fluorescence Lifetime Imaging Microscopy Datasets, PLOS ONE, Vol: 8, ISSN: 1932-6203
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- Citations: 147
Seidenari S, Arginelli F, Dunsby C, et al., 2013, Multiphoton Laser Tomography and Fluorescence Lifetime Imaging of Melanoma: Morphologic Features and Quantitative Data for Sensitive and Specific Non-Invasive Diagnostics, PLOS ONE, Vol: 8, ISSN: 1932-6203
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- Citations: 59
Alibhai D, Kelly DJ, Warren S, et al., 2013, Automated fluorescence lifetime imaging plate reader and its application to Forster resonant energy transfer readout of Gag protein aggregation, Journal of Biophotonics, Vol: 6, Pages: 398-408, ISSN: 1864-0648
Fluorescence lifetime measurements can provide quantitativereadouts of local fluorophore environment andcan be applied to biomolecular interactions via Fo¨ rsterresonant energy transfer (FRET). Fluorescence lifetimeimaging (FLIM) can therefore provide a high contentanalysis (HCA) modality to map protein-protein interactions(PPIs) with applications in drug discovery, systemsbiology and basic research. We present here an automatedmultiwell plate reader able to perform rapid unsupervisedoptically sectioned FLIM of fixed and livebiological samples and illustrate its potential to assayPPIs through application to Gag protein aggregationduring the HIV life cycle. We demonstrate both heteroFRETand homo-FRET readouts of protein aggregationand report the first quantitative evaluation of a FLIMHCA assay by generating dose response curves throughaddition of an inhibitor of Gag myristoylation. Z0 factorsexceeding 0.6 are realised for this FLIM FRET assay.Fluorescence lifetime plate map with representativeimages of high and low FRET cells and correspondingdose response plot.
Arginelli F, Manfredini M, Bassoli S, et al., 2013, High resolution diagnosis of common nevi by multiphoton laser tomography and fluorescence lifetime imaging, SKIN RESEARCH AND TECHNOLOGY, Vol: 19, Pages: 194-204, ISSN: 0909-752X
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- Citations: 9
Chen L, Andrews N, Kumar S, et al., 2013, Simultaneous angular multiplexing optical projection tomography at shifted focal planes, OPTICS LETTERS, Vol: 38, Pages: 851-853, ISSN: 0146-9592
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- Citations: 16
Manfredini M, Arginelli F, Dunsby C, et al., 2013, High-resolution imaging of basal cell carcinoma: a comparison between multiphoton microscopy with fluorescence lifetime imaging and reflectance confocal microscopy, SKIN RESEARCH AND TECHNOLOGY, Vol: 19, Pages: E433-E443, ISSN: 0909-752X
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- Citations: 21
Coda S, Kelly DJ, Lagarto JL, et al., 2013, Autofluorescence lifetime imaging and metrology for medical research and clinical diagnosis
We report the development of instrumentation to utilise autofluorescence lifetime for the study and diagnosis of disease including cancer and osteoarthritis. ©2013 The Optical Society (OSA).
Kelly DJ, Alibhai D, Warren S, et al., 2013, An automated flim multiwell plate reader for high content analysis
We report an automated fluorescence lifetime imaging multiwell plate reader for high content analysis, capable of subcellular mapping of protein interactions. This instrument can acquire FLIM data from 96 wells in less than 15 minutes.©2013 The Optical Society (OSA).
Warren S, Kimberley C, Margineanu A, et al., 2013, Flim-fret of cell signalling in chemotaxis
We demonstrate the application of Fluorescence Lifetime Imaging (FLIM) to read out Förster resonant energy transfer (FRET) based biosensors for studying the spatio-temporal dynamics of signalling pathways in cells undergoing chemotaxis. ©2013 The Optical Society (OSA).
Manning HB, Nickdel MB, Yamamoto K, et al., 2013, Detection of cartilage matrix degradation by autofluorescence lifetime, MATRIX BIOLOGY, Vol: 32, Pages: 32-38, ISSN: 0945-053X
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- Citations: 31
Roper JC, Yerolatsitis S, Birks TA, et al., 2013, Minimising group index variations in a multicore endoscope fibre, Conference on Lasers and Electro-Optics (CLEO), Publisher: IEEE, ISSN: 2160-9020
Martins M, Warren S, Kimberley C, et al., 2012, Activity of PLCε contributes to chemotaxis of fibroblasts towards PDGF, JOURNAL OF CELL SCIENCE, Vol: 125, Pages: 5758-5769, ISSN: 0021-9533
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- Citations: 15
Laine R, Stuckey DW, Manning H, et al., 2012, Fluorescence Lifetime Readouts of Troponin-C-Based Calcium FRET Sensors: A Quantitative Comparison of CFP and mTFP1 as Donor Fluorophores, PLOS ONE, Vol: 7, ISSN: 1932-6203
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- Citations: 20
Seidenari S, Arginelli F, Dunsby C, et al., 2012, Multiphoton laser tomography and fluorescence lifetime imaging of basal cell carcinoma: morphologic features for non-invasive diagnostics, EXPERIMENTAL DERMATOLOGY, Vol: 21, Pages: 831-836, ISSN: 0906-6705
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- Citations: 32
Xavier GDS, Mondragon A, Sun G, et al., 2012, Abnormal glucose tolerance and insulin secretion in pancreas-specific <i>Tcf7l2</i>-null mice, DIABETOLOGIA, Vol: 55, Pages: 2667-2676, ISSN: 0012-186X
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- Citations: 69
Patalay R, Talbot C, Alexandrov Y, et al., 2012, Multiphoton Multispectral Fluorescence Lifetime Tomography for the Evaluation of Basal Cell Carcinomas, PLOS One, Vol: 7, ISSN: 1932-6203
We present the first detailed study using multispectral multiphoton fluorescence lifetime imaging to differentiate basal cell carcinoma cells (BCCs) from normal keratinocytes. Images were acquired from 19 freshly excised BCCs and 27 samples of normal skin (in & ex vivo). Features from fluorescence lifetime images were used to discriminate BCCs with a sensitivity/specificity of 79%/93% respectively. A mosaic of BCC fluorescence lifetime images covering >1 mm2 is also presented, demonstrating the potential for tumour margin delineation.Using 10,462 manually segmented cells from the image data, we quantify the cellular morphology and spectroscopic differences between BCCs and normal skin for the first time. Statistically significant increases were found in the fluorescence lifetimes of cells from BCCs in all spectral channels, ranging from 19.9% (425–515 nm spectral emission) to 39.8% (620–655 nm emission). A discriminant analysis based diagnostic algorithm allowed the fraction of cells classified as malignant to be calculated for each patient. This yielded a receiver operator characteristic area under the curve for the detection of BCC of 0.83.We have used both morphological and spectroscopic parameters to discriminate BCC from normal skin, and provide a comprehensive base for how this technique could be used for BCC assessment in clinical practice.
Brown AC, Oddos S, Dobbie IM, et al., 2012, Correction: Remodelling of Cortical Actin Where Lytic Granules Dock at Natural Killer Cell Immune Synapses Revealed by Super-Resolution Microscopy., PLoS Biol, Vol: 10
[This corrects the article on p. e1001152 in vol. 9.].
Patalay R, Chu A, Dunsby C, et al., 2012, A noninvasive imaging study of skin using two photon microscopy of cellular autofluorescence, 70th Annual Meeting of the American-Academy-of-Dermatology (AAD), Publisher: MOSBY-ELSEVIER, Pages: AB83-AB83, ISSN: 0190-9622
Chen L, McGinty J, Taylor HB, et al., 2012, Incorporation of an experimentally determined MTF for spatial frequency filtering and deconvolution during optical projection tomography reconstruction, OPTICS EXPRESS, Vol: 20, Pages: 7323-7337, ISSN: 1094-4087
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- Citations: 15
Thompson AJ, Coda S, Sorensen MB, et al., 2012, In vivo measurements of diffuse reflectance and time-resolved autofluorescence emission spectra of basal cell carcinomas, JOURNAL OF BIOPHOTONICS, Vol: 5, Pages: 240-254, ISSN: 1864-063X
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- Citations: 30
Sardini A, Stuckey DW, McGinty J, et al., 2012, In Vivo Investigation of Calpain Activity by Lifetime Imaging of Genetically Encoded FRET Sensors, BIOPHYSICAL JOURNAL, Vol: 102, Pages: 159A-159A, ISSN: 0006-3495
Chen L, McGinty J, Taylor HB, et al., 2012, Improved OPT reconstructions based on the MTF and extension to FLIM-OPT
We demonstrate the improved reconstruction of OPT datasets by incorporating the measured MTF in the reconstruction process. We also extend OPT to FLIM-OPT and demonstrate its use for imaging live zebrafish embryos displaying autofluorescence. © 2012 OSA.
Esseling M, Kemper B, Antkowiak M, et al., 2012, Multimodal biophotonic workstation for live cell analysis, JOURNAL OF BIOPHOTONICS, Vol: 5, Pages: 9-13, ISSN: 1864-063X
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- Citations: 18
Antkowiak M, Torres-Mapa ML, McGinty J, et al., 2012, Towards gene therapy based on femtosecond optical transfection, BIOPHOTONICS: PHOTONIC SOLUTIONS FOR BETTER HEALTH CARE III, Vol: 8427, ISSN: 0277-786X
Seidenari S, Arginelli F, Bassoli S, et al., 2012, Multiphoton laser microscopy and fluorescence lifetime imaging for the evaluation of the skin., Dermatol Res Pract, Vol: 2012
Multiphoton laser microscopy is a new, non-invasive technique providing access to the skin at a cellular and subcellular level, which is based both on autofluorescence and fluorescence lifetime imaging. Whereas the former considers fluorescence intensity emitted by epidermal and dermal fluorophores and by the extra-cellular matrix, fluorescence lifetime imaging (FLIM), is generated by the fluorescence decay rate. This innovative technique can be applied to the study of living skin, cell cultures and ex vivo samples. Although still limited to the clinical research field, the development of multiphoton laser microscopy is thought to become suitable for a practical application in the next few years: in this paper, we performed an accurate review of the studies published so far, considering the possible fields of application of this imaging method and providing high quality images acquired in the Department of Dermatology of the University of Modena.
Soloviev VY, McGinty J, Stuckey DW, et al., 2011, Förster resonance energy transfer imaging in vivo with approximated radiative transfer equation, Applied Optics, Vol: 50, Pages: 6583-6590
We describe a new light transport model, which was applied to three-dimensional lifetime imaging of Förster resonance energy transfer in mice in vivo. The model is an approximation to the radiative transfer equation and combines light diffusion and ray optics. This approximation is well adopted to wide-field time-gated intensity-based data acquisition. Reconstructed image data are presented and compared with results obtained by using the telegraph equation approximation. The new approach provides improved recovery of absorption and scattering parameters while returning similar values for the fluorescence parameters.
Patalay R, Talbot C, Alexandrov Y, et al., 2011, Non-invasive imaging of skin cancer with fluorescence lifetime imaging using two photon tomography, Optics InfoBase Conference Papers
Multispectral fluorescence lifetime imaging (FLIM) using two photon microscopy as a non-invasive technique for the diagnosis of skin lesions is described. Skin contains fluorophores including elastin, keratin, collagen, FAD and NADH. This endogenous contrast allows tissue to be imaged without the addition of exogenous agents and allows the in vivo state of cells and tissues to be studied. A modified DermaInspect® multiphoton tomography system was used to excite autofluorescence at 760 nm in vivo and on freshly excised ex vivo tissue. This instrument simultaneously acquires fluorescence lifetime images in four spectral channels between 360-655 nm using time-correlated single photon counting and can also provide hyperspectral images. The multispectral fluorescence lifetime images were spatially segmented and binned to determine lifetimes for each cell by fitting to a double exponential lifetime model. A comparative analysis between the cellular lifetimes from different diagnoses demonstrates significant diagnostic potential. © 2011 SPIE-OSA.
Patalay R, Talbot C, Alexandrov Y, et al., 2011, Quantification of cellular autofluorescence of human skin using multiphoton tomography and fluorescence lifetime imaging in two spectral detection channels, Biomedical Optics Express, Vol: 2, Pages: 3295-3308, ISSN: 2156-7085
We explore the diagnostic potential of imaging endogenous fluorophores using two photon microscopy and fluorescence lifetime imaging (FLIM) in human skin with two spectral detection channels. Freshly excised benign dysplastic nevi (DN) and malignant nodular Basal Cell Carcinomas (nBCCs) were excited at 760 nm. The resulting fluorescence signal was binned manually on a cell by cell basis. This improved the reliability of fitting using a double exponential decay model and allowed the fluorescence signatures from different cell populations within the tissue to be identified and studied. We also performed a direct comparison between different diagnostic groups. A statistically significant difference between the median mean fluorescence lifetime of 2.79 ns versus 2.52 ns (blue channel, 300-500 nm) and 2.08 ns versus 1.33 ns (green channel, 500-640 nm) was found between nBCCs and DN respectively, using the Mann-Whitney U test (p < 0.01). Further differences in the distribution of fluorescence lifetime parameters and inter-patient variability are also discussed.
Brown ACN, Oddos S, Dobbie IM, et al., 2011, Remodelling of cortical actin where lytic granules dock at natural killer cell immune synapses revealed by super-resolution microscopy, PLoS Biology, Vol: 9, Pages: 1-18, ISSN: 1544-9173
Natural Killer (NK) cells are innate immune cells that secrete lytic granules to directly kill virus-infected or transformed cells across an immune synapse. However, a major gap in understanding this process is in establishing how lytic granules pass through the mesh of cortical actin known to underlie the NK cell membrane. Research has been hampered by the resolution of conventional light microscopy, which is too low to resolve cortical actin during lytic granule secretion. Here we use two high-resolution imaging techniques to probe the synaptic organisation of NK cell receptors and filamentous (F)-actin. A combination of optical tweezers and live cell confocal microscopy reveals that microclusters of NKG2D assemble into a ring-shaped structure at the centre of intercellular synapses, where Vav1 and Grb2 also accumulate. Within this ring-shaped organisation of NK cell proteins, lytic granules accumulate for secretion. Using 3D-structured illumination microscopy (3D-SIM) to gain super-resolution of ∼100 nm, cortical actin was detected in a central region of the NK cell synapse irrespective of whether activating or inhibitory signals dominate. Strikingly, the periodicity of the cortical actin mesh increased in specific domains at the synapse when the NK cell was activated. Two-colour super-resolution imaging revealed that lytic granules docked precisely in these domains which were also proximal to where the microtubule-organising centre (MTOC) polarised. Together, these data demonstrate that remodelling of the cortical actin mesh occurs at the central region of the cytolytic NK cell immune synapse. This is likely to occur for other types of cell secretion and also emphasises the importance of emerging super-resolution imaging technology for revealing new biology.
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