Publications
300 results found
Coda S, Kennedy GT, Thompson A, et al., 2011, FLUORESCENCE LIFETIME IMAGING FOR LABEL-FREE CONTRAST OF GASTROINTESTINAL DISEASES, International School of Physics "Enrico Fermi", Course CLXXXI "Microscopy Applied to Biophotonics"
INTRODUCTION: Autofluorescence (AF) has been used to distinguish between normal and diseased tissue, but its molecular basis is still unclear and making quantitative intensity measurements is challenging. Fluorescence lifetime imaging (FLIM) measures the decay rate of the autofluorescent signal from tissue, providing quantitative AF contrast. FLIM has been recently implemented by our group in three endoscopic instruments consisting of a confocal laser endomicroscope, a wide-field fibre-optic endoscope and a single point fibre-optic probe. FLIM has the potential to report on tissue structure and function in real-time during endoscopy, providing a label-free means to detect the early onset of diseases that cause changes in tissue AF. We are developing these 3 modalities for in vivo clinical application, supported by ex vivo studies on freshly-biopsied/resected GI tissues.AIMS & METHODS: The aim of this work is to translate our FLIM instrumentation from the optical bench to in vivo clinical application. AF from 43 endoscopic samples from different GI sites was excited using a conventional confocal FLIM microscope in the range 405-420nm, which is compatible with our FLIM endoscopes, and which is the range needed to excite a number of important endogenous GI tissue fluorophores such as porphyrins, flavins, collagen and elastin. The samples were collected from patients undergoing endoscopy, transported to the FLIM laboratory to be imaged and then submitted for histopathology. The following disorders were investigated: Barrett’s oesophagus, gastric cancer, ulcerative colitis, Crohn’s disease, adenomatous polyps and colon cancer. The accuracy of FLIM in discriminating dysplastic/cancerous samples from normal tissue has been tested by measuring the Area Under the Curve (AUC).RESULTS: Our preliminary data show that premalignant or neoplastic samples display either shorter or longer fluorescence lifetime than that of normal tissue. Increased lifetime val
Boruah BR, Love GD, Neil MAA, 2011, Interferometry using binary holograms without high order diffraction effects, OPTICS LETTERS, Vol: 36, Pages: 2357-2359, ISSN: 0146-9592
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- Citations: 15
Coda S, Kennedy G, Thompson A, et al., 2011, FLUORESCENCE LIFETIME IMAGING OF GASTROINTESTINAL CANCERS, European-Society-for-Medical-Oncology (ESMO) 13th World Congress on Gastrointestinal Cancer, Publisher: OXFORD UNIV PRESS, Pages: v65-v66, ISSN: 0923-7534
Thompson AJ, Paterson C, Neil MAA, et al., 2011, Adaptive phase compensation for ultracompact laser scanning endomicroscopy, OPTICS LETTERS, Vol: 36, Pages: 1707-1709, ISSN: 0146-9592
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- Citations: 68
Grippon S, Zhao Q, Robinson T, et al., 2011, Differential modes of DNA binding by mismatch uracil DNA glycosylase from Escherichia coli: implications for abasic lesion processing and enzyme communication in the base excision repair pathway, Nucleic Acids Research, Vol: 39, Pages: 2593-2603, ISSN: 1362-4962
Mismatch uracil DNA glycosylase (Mug) fromEscherichia coli is an initiating enzyme in thebase-excision repair pathway. As with other DNAglycosylases, the abasic product is potentiallymore harmful than the initial lesion. Since Mug isknown to bind its product tightly, inhibitingenzyme turnover, understanding how Mug bindsDNA is of significance when considering how Muginteracts with downstream enzymes in the baseexcisionrepair pathway. We have demonstrateddifferential binding modes of Mug between its substrateand abasic DNA product using both band shiftand fluorescence anisotropy assays. Mug binds itsproduct cooperatively, and a stoichiometric analysisof DNA binding, catalytic activity and saltdependenceindicates that dimer formation is offunctional significance in both catalytic activity andproduct binding. This is the first report ofcooperativity in the uracil DNA glycosylase superfamilyof enzymes, and forms the basis of productinhibition in Mug. It therefore provides a new perspectiveon abasic site protection and the findingsare discussed in the context of downstream lesionprocessing and enzyme communication in the baseexcision repair pathway.
Everett KL, Buehler A, Bunney TD, et al., 2011, Membrane Environment Exerts an Important Influence on Rac-Mediated Activation of Phospholipase Cγ2, MOLECULAR AND CELLULAR BIOLOGY, Vol: 31, Pages: 1240-1251, ISSN: 0270-7306
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- Citations: 21
Salehi-Reyhani A, Kaplinsky J, Burgin E, et al., 2011, A first step towards practical single cell proteomics: a microfluidic antibodycapture chip with TIRF detection, Lab Chip, Vol: 11, Pages: 1256-1261
We have developed a generic platform to undertake the analysis of protein copy number from singlecells. The approach described here is ‘all-optical’ whereby single cells are manipulated into separate analysis chambers using an optical trap; single cells are lysed by a shock wave caused by laser-induced microcavitation, and the protein released from a single cell is measured by total internal reflection microscopy as it is bound to micro-printed antibody spots within the device. The platform was tested using GFP transfected cells and the relative precision of the measurement method was determined to be 88%. Single cell measurements were also made on a breast cancer cell line to measure the relative levels of unlabelled human tumour suppressor protein p53 using a chip incorporating an antibody sandwich assay format. These results suggest that this is a viable method for measuring relative protein levels in single cells.
Kumar S, Alibhai D, Margineanu A, et al., 2011, FLIM FRET technology for drug discovery: automated multiwell-plate high-content analysis, multiplexed readouts and application in situ, ChemPhysChem: a European journal of chemical physics and physical chemistry, Vol: 12, Pages: 609-626, ISSN: 1439-4235
A fluorescence lifetime imaging (FLIM) technology platform intendedto read out changes in Fçrster resonance energy transfer(FRET) efficiency is presented for the study of protein interactionsacross the drug-discovery pipeline. FLIM provides arobust, inherently ratiometric imaging modality for drug discoverythat could allow the same sensor constructs to betranslated from automated cell-based assays through smalltransparent organisms such as zebrafish to mammals. To thisend, an automated FLIM multiwell-plate reader is described forhigh content analysis of fixed and live cells, tomographic FLIMin zebrafish and FLIM FRET of live cells via confocal endomicroscopy.For cell-based assays, an exemplar application readingout protein aggregation using FLIM FRET is presented, andthe potential for multiple simultaneous FLIM (FRET) readoutsin microscopy is illustrated.
Galletly N, McGinty J, Munro I, et al., 2011, Fluorescence lifetime imaging of liver cancer, 107th Annual Meeting of the American-Gastroenterlogical Association, Publisher: W B Saunders Co-Elsevier Inc
Margineanu A, Laine R, Kumar S, et al., 2011, Multiplexed Time Lapse Fluorescence Lifetime Readouts in an Optically Sectioning Time-Gated Imaging Microscope, 55th Annual Meeting of the Biophysical-Society, Publisher: CELL PRESS, Pages: 183-183, ISSN: 0006-3495
Thompson A, Manning H, Brydegaard M, et al., 2011, Hyperspectral fluorescence lifetime fibre probe spectroscopy for use in the study and diagnosis of osteoarthritis and skin cancer, SPIE Photonics West 2011, Publisher: Society of Photo-optical Instrumentation Engineers (SPIE), ISSN: 1996-756X
We present the application of two fibre-optic-coupled time-resolved spectrofluorometers and a compact steady-state diffuse reflected light/fluorescence spectrometer to in vivo and ex vivo studies of skin cancer and osteoarthritis. In a clinical study of skin cancer, 27 lesions on 25 patients were investigated in vivo before surgical excision of the region measured. Preliminary analysis reveals a statistically significant decrease in the autofluorescence lifetime of basal cell carcinomas compared to neighbouring healthy tissue. A study of autofluorescence signals associated with the onset of osteoarthritis indicates autofluorescence lifetime changes associated with collagen degradation.
McGovern B, Drakakis E, Neil M, et al., 2011, Individually addressable optoelectronic arrays for optogenetic neural stimulation, Annual IEEE Biomedical Circuits and Systems Conference (BioCAS) - Engineering Tomorrow's Healthcare, Publisher: IEEE, Pages: 329-332, ISSN: 2163-4025
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- Citations: 4
McGinty J, Talbot C, Owen D, et al., 2011, Fluorescence Lifetime Imaging Microscopy, Endoscopy and Tomography, Editors: Boas, Pitris, Ramanujam, ISBN: 1420090364
Talbot CB, Patalay R, Munro I, et al., 2011, A multispectral FLIM tomograph for <i>in vivo</i> imaging of skin cancer, Conference on Multiphoton Microscopy in the Biomedical Sciences XI, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
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- Citations: 3
Lenz MO, Brown ACN, Auksorius E, et al., 2011, A STED-FLIM microscope applied to imaging the Natural Killer cell immune synapse, Conference on Multiphoton Microscopy in the Biomedical Sciences XI, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
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- Citations: 5
Patalay R, Talbot C, Alexandrov Y, et al., 2011, Non-invasive imaging of skin cancer with fluorescence lifetime imaging using two photon tomography, Conference on Clinical and Biomedical Spectroscopy and Imaging II, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
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- Citations: 3
Alibhai D, Kumar S, Kelly D, et al., 2011, An automated wide-field, time-gated, optically sectioning, fluorescence lifetime imaging multiwell plate reader for high-content analysis of protein-protein interactions, Conference on Three-Dimensional and Multidimensional Microscopy - Image Acquisition and Processing XVIII, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Manivannan N, Neil MAA, 2011, Automatic angle measurement of a 2D object using optical correlator-neural networks hybrid system, Conference on Optical Pattern Recognition XXII, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Kennedy GT, Coda S, Thompson AJ, et al., 2011, Fluorescence Lifetime Imaging Endoscopy, Conference on Endoscopic Microscopy VI, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
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- Citations: 3
Manivannan N, Neil MAA, 2010, Optical correlator-neural network hybrid system for many patterns recognition, Pages: 249-251
In this paper it is proposed and demonstrated optical correlator-neural network architecture for many pattern recognition. An intelligent training scheme is employed to train the hybrid system so that similarities in the input patterns are used efficiently. Both computer simulation and experimental results are presented for recognition of characters from the English alphabetic. ©2010 IEEE.
McGinty J, Stuckey D, Laine R, et al., 2010, Time-domain fluorescence lifetime optical projection tomography
We present a platform for measuring the fluorescence lifetime distribution in mesoscopic samples (~0.1-1cm) based on optical projection tomography and time-gated imaging. This is applied to optically cleared embryos expressing a calcium sensing FRET probe. © OSA / BIOMED/DH 2010.
McGovern B, Berlinguer-Palmini R, Grossman N, et al., 2010, A new individually addressable micro-LED array for photogenetic neural stimulation, IEEE Trans. Biomedical Circuits and Systems, Vol: In press
Purbhoo MA, Liu H, Oddos S, et al., 2010, Dynamics of sub-synaptic vesicles and surface microclusters at the T cell immunological synapse, Annual Congress of the British-Society-for-Immunology, Publisher: WILEY-BLACKWELL PUBLISHING, INC, Pages: 175-175, ISSN: 0019-2805
McGovern B, Palmini RB, Grossman N, et al., 2010, A New Individually Addressable Micro-LED Array for Photogenetic Neural Stimulation, IEEE Transactions on Biomedical Circuits and Systems, Vol: 4, Pages: 469-476, ISSN: 1932-4545
McGinty J, Galletly NP, Dunsby C, et al., 2010, Wide-field fluorescence lifetime imaging of cancer, BIOMEDICAL OPTICS EXPRESS, Vol: 1, Pages: 627-640, ISSN: 2156-7085
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- Citations: 77
Owen DM, Oddos S, Kumar S, et al., 2010, High plasma membrane lipid order imaged at the immunological synapse periphery in live T cells, MOLECULAR MEMBRANE BIOLOGY, Vol: 27, Pages: 178-189, ISSN: 0968-7688
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- Citations: 64
Purbhoo MA, Liu H, Oddos S, et al., 2010, Dynamics of Subsynaptic Vesicles and Surface Microclusters at the Immunological Synapse, SCIENCE SIGNALING, Vol: 3, ISSN: 1945-0877
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- Citations: 101
McGovern BP, Palmini RB, Grossman N, et al., 2010, Towards an Optogenetic Retinal Prosthesis, Publisher: ASSOC RESEARCH VISION OPHTHALMOLOGY INC, ISSN: 0146-0404
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- Citations: 1
Grossman N, Poher V, Grubb MS, et al., 2010, Multisite optical excitation using ChR2 and micro-LED array, Journal of Neural Engineering, Vol: 7, ISSN: 1741-2560
McGinty J, Stuckey D, Laine R, et al., 2010, Time-domain fluorescence lifetime optical projection tomography
We present a platform for measuring the fluorescence lifetime distribution in mesoscopic samples (~0.1-1cm) based on optical projection tomography and time-gated imaging. This is applied to optically cleared embryos expressing a calcium sensing FRET probe. © OSA / BIOMED/DH 2010.
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