481 results found
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
McGinty J, Taylor HB, Chen L, et al., 2011, In vivo fluorescence lifetime optical projection tomography, Biomedical Optics Express, Vol: 2, Pages: 1340-1350, ISSN: 2156-7085
We demonstrate the application of fluorescence lifetime optical projection tomography (FLIM-OPT) to in vivo imaging of lysC:GFP transgenic zebrafish embryos (Danio rerio). This method has been applied to unambiguously distinguish between the fluorescent protein (GFP) signal in myeloid cells from background autofluorescence based on the fluorescence lifetime. The combination of FLIM, an inherently ratiometric method, in conjunction with OPT results in a quantitative 3-D tomographic technique that could be used as a robust method for in vivo biological and pharmaceutical research, for example as a readout of Förster resonance energy transfer based interactions.
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
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 C gamma 2, MOLECULAR AND CELLULAR BIOLOGY, Vol: 31, Pages: 1240-1251, ISSN: 0270-7306
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, Vol: 12, Pages: 609-626
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.
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.
Ushakov DS, Caorsi V, Ibanez-Garcia D, et al., 2011, Response of Rigor Cross-bridges to Stretch Detected by Fluorescence Lifetime Imaging Microscopy of Myosin Essential Light Chain in Skeletal Muscle Fibers, JOURNAL OF BIOLOGICAL CHEMISTRY, Vol: 286, Pages: 842-850, ISSN: 0021-9258
Talbot CB, Patalay R, Munro I, et al., 2011, A multispectral FLIM tomograph for in vivo imaging of skin cancer, Conference on Multiphoton Microscopy in the Biomedical Sciences XI, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
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
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
French PMW, 2011, Fluorescence lifetime imaging: FLIM for cell biology, drug discovery and label-free diagnosis
I will present FLIM technology to read out biomolecular interactions across the scales from labelled proteins in solution and in cells through automated plate readers to imaging disease models and endoscopic diagnosis using autofluorescence. © 2011 OSA: BODA/NTM/OMP/OTA.
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
McGinty J, Talbot C, Owen D, et al., 2011, Fluorescence Lifetime Imaging Microscopy, Endoscopy and Tomography, Editors: Boas, Pitris, Ramanujam, ISBN: 1420090364
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.
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
Ibanez-Garcia D, Requejo-Isidro J, Webb MR, et al., 2010, Fluorescence Lifetime Imaging Reveals that the Environment of the ATP Binding Site of Myosin in Muscle Senses Force, BIOPHYSICAL JOURNAL, Vol: 99, Pages: 2163-2169, ISSN: 0006-3495
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
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
Sun G, Tarasov AI, McGinty JA, et al., 2010, LKB1 deletion with the RIP2.Cre transgene modifies pancreatic beta-cell morphology and enhances insulin secretion in vivo, AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM, Vol: 298, Pages: E1261-E1273, ISSN: 0193-1849
Talbot C, McGinty J, McGhee E, et al., 2010, Fluorescence Lifetime Imaging and Metrology for Biomedicine, Handbook of Photonics for Biomedical Science, ISBN: 978-1439806289
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
Sun G, Tarasov AI, McGinty J, et al., 2010, Ablation of AMP-activated protein kinase alpha 1 and alpha 2 from mouse pancreatic beta cells and RIP2.Cre neurons suppresses insulin release in vivo, DIABETOLOGIA, Vol: 53, Pages: 924-936, ISSN: 0012-186X
Soloviev VY, McGinty J, Tahir KB, et al., 2010, Tomographic imaging of fluorescence resonance energy transfer in highly scattering media, SPIE Photonics West, Publisher: SPIE, ISSN: 1605-7422
McGinty J, Stuckey DW, Tahir KB, et al., 2010, tomoFLIM - fluorescence lifetime projection tomography, Conference on Three-Dimensional and Multidimensional Microscopy - Image Acquisition and Processing XVII, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Robinson T, Manning HB, Dunsby C, et al., 2010, Investigating fast enzyme-DNA kinetics using multidimensional fluorescence imaging and microfluidics, Conference on Microfluidics, BioMEMS, and Medical Microsystems VIII, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Soloviev VY, McGinty J, Tahir KB, et al., 2010, Tomographic imaging of Fluorescence Resonance Energy Transfer in highly light scattering media, Conference on Biomedical Applications of Light Scattering IV, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
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