482 results found
Manning HB, Owen DM, Auksorius E, et al., 2007, Applications of rapid time-gated hyperspectral FLIM: Live cell imaging of membrane order and 6-D microscopy, ISSN: 1605-7422
We describe the characterisation of a hyperspectral fluorescence lifetime imaging microscope that exploits high-speed time-gated imaging technology and a tunable continuum source for 6-D fluorescence imaging. This line-scanning confocal microscope can record the full spectral-temporal (i.e. excitation-emission-lifetime) fluorescence matrix at each pixel in a three dimensional (x-y-z) sample. This instrument has been applied to biological samples including model membranes and live cells labelled with the phase-sensitive membrane dye di-4-ANEPPDHQ, for which significant variation of lifetime with emission wavelength is observed. © 2007 SPIE-OSA.
Grant DM, McGinty J, McGhee EJ, et al., 2007, High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events, Optics Express, Vol: 15, Pages: 16656-16673
We present a time domain optically sectioned fluorescence lifetime imaging (FLIM) microscope developed for high-speed live cell imaging. This single photon excited system combines wide field parallel pixel detection with confocal sectioning utilizing spinning Nipkow disc microscopy. It can acquire fluorescence lifetime images of live cells at up to 10 frames per second (fps), permitting high-speed FLIM of cell dynamics and protein interactions with potential for high throughput cell imaging and screening applications. We demonstrate the application of this FLIM microscope to real-time monitoring of changes in lipid order in cell membranes following cholesterol depletion using cyclodextrin and to the activation of the small GTP-ase Ras in live cells using FRET. © 2007 Optical Society of America.
Grant DM, McGinty J, McGhee EJ, et al., 2007, High speed optically sectioned fluorescence lifetime imaging permits study of live cell signaling events, OPTICS EXPRESS, Vol: 15, Pages: 15656-15673, ISSN: 1094-4087
Benninger RK, Hofmann O, Onfelt B, et al., 2007, Fluorescence-Lifetime Imaging of DNA-Dye Interactions within Continuous-Flow Microfluidic Systems., Angew Chem Int Ed Engl, Vol: 46, Pages: 8536-8536, ISSN: 1433-7851
Soloviev VY, Tahir KB, McGinty J, et al., 2007, Fluorescence lifetime imaging by using time-gated data acquisition, APPLIED OPTICS, Vol: 46, Pages: 7384-7391, ISSN: 1559-128X
Gong Z, Zhang HX, Gu E, et al., 2007, Matrix-addressable micropixellated InGaN light-emitting diodes with uniform emission and increased light output, IEEE TRANSACTIONS ON ELECTRON DEVICES, Vol: 54, Pages: 2650-2658, ISSN: 0018-9383
Owen DM, Neil MAA, French PMW, et al., 2007, Optical techniques for imaging membrane lipid microdomains in living cells, SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY, Vol: 18, Pages: 591-598, ISSN: 1084-9521
Kumar S, Dunsby C, De Beule PAA, et al., 2007, Multifocal multiphoton excitation and time correlated single photon counting detection for 3-D fluorescence lifetime imaging, Optics Express, Vol: 15, Pages: 12548-12561
We report a multifocal multiphoton time-correlated single photon counting (TCSPC) fluorescence lifetime imaging (FLIM) microscope system that uses a 16 channel multi-anode PMT detector. Multiphoton excitation minimizes out-of-focus photobleaching, multifocal excitation reduces non-linear in-plane photobleaching effects and TCSPC electronics provide photon-efficient detection of the fluorescence decay profile. TCSPC detection is less prone to bleaching- and movementinduced artefacts compared to wide-field time-gated or frequency-domain FLIM. This microscope is therefore capable of acquiring 3-D FLIM images at significantly increased speeds compared to single beam multiphoton microscopy and we demonstrate this with live cells expressing a GFP tagged protein. We also apply this system to time-lapse FLIM of NAD(P)H autofluorescence in single live cells and report measurements on the change in the fluorescence decay profile following the application of a known metabolic inhibitor.
Poher V, Zhang HX, Kennedy GT, et al., 2007, Optical sectioning microscopes with no moving parts using a micro-stripe array light emitting diode, OPTICS EXPRESS, Vol: 15, Pages: 11196-11206, ISSN: 1094-4087
Garcia DI, Lanigan P, Webb M, et al., 2007, Fluorescence lifetime imaging to detect actomyosin states in mammalian muscle sarcomeres, BIOPHYSICAL JOURNAL, Vol: 93, Pages: 2091-2101, ISSN: 0006-3495
Soloviev VY, McGinty J, Tahir KB, et al., 2007, Fluorescence lifetime tomography of live cells expressing enhanced green fluorescent protein embedded in a scattering medium exhibiting background autofluorescence, OPTICS LETTERS, Vol: 32, Pages: 2034-2036, ISSN: 0146-9592
Soloviev VY, Tahir KB, McGinty J, et al., 2007, Fluorescence lifetime imaging through turbid media reconstructed in the Fourier domain using time-gated imaging data, European Conference on Biomedical Optics, Publisher: SPIE
Hegyi L, Talbot C, Monaco C, et al., 2007, Fluorescence lifetime imaging of unstained human atherosclerotic plaques., The 14th International Symposium on Atherosclerosis
Kennedy GT, Elson DS, Munro I, et al., 2007, Fluorescence lifetime imaging using light emitting diodes, Conference on Three-Dimensional and Multidimensional Microscopy - Image Acquisition and Processing XIV, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Ushakov DS, Konitsiotis A, Garcia DI, et al., 2007, Investigation of myosin essential light chain in skeletal muscle fibres by fluorescence lifetime imaging microscopy., 51st Annual Meeting of the Biophysical-Society, Publisher: BIOPHYSICAL SOCIETY, Pages: 296A-296A, ISSN: 0006-3495
Benninger RKP, Hofmann O, Onfelt B, et al., 2007, Fluorescence-lifetime imaging of DNA-dye interactions within continuous-flow microfluidic systems, ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, Vol: 46, Pages: 2228-2231, ISSN: 1433-7851
Owen DM, Manning HB, de Beule P, et al., 2007, Development of a hyperspectral fluorescence lifetime imaging microscope and its application to tissue imaging, Conference on Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues V, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Poher V, Kennedy GT, Elson DS, et al., 2007, Microscopy using micro-pixellated light emitting diodes, Conference on Three-Dimensional and Multidimensional Microscopy - Image Acquisition and Processing XIV, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Manning HB, Owen DM, Auksorius E, et al., 2007, Applications of rapid time-gated hyperspectral FLIM: live cell imaging of membrane order and 6-D microscopy, Conference on Confocal, Multiphoton and Nonlinear Microscopic Imaging III, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Soloviev VY, Tahir KB, McGinty J, et al., 2007, Fluorescence lifetime imaging through turbid media reconstructed in the Fourier domain using time-gated imaging data, Conference on Diffuse Optical Imaging of Tissue, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Benninger RKP, Hofmann O, Onfelt B, et al., 2007, Fluorescence-lifetime imaging of DNA-Dye interactions within continuous-flow microfluidic systems (vol 46, pg 8525, 2007), ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, Vol: 46, Pages: 8536-8536, ISSN: 1433-7851
Poher V, Kennedy G T, Elson D S, et al., 2007, Microscopy using micropixellated light emitting diodes, THREE-DIMENSIONAL AND MULTIDIMENSIONAL MICROSCOPY: IMAGE ACQUISITION AND PROCESSING XIV, BIOS
De Beule PAA, Dunsby C, Owen DM, et al., 2007, A novel hyperspectral lifetime probe for autofluorescence - art. no. 643303, Conference on Optical Fibers and Sensors for Medical Diagnostics and Treatment Applications VII, Pages: 43303-43303
The application of autofluorescence in non-invasive medical diagnostics could have great potential. Two major drawbacks inherent to this approach are low signal levels compared to those from exogenous fluorescent probes and complexity caused by the multiplicity of fluorescent biomolecules in tissue. Here we present a new optical system that is based on single channel detection via all optical fiber and call measure the fluorescence emission spectrum and fluorescence lifetime simultaneously for excitation wavelengths of 355 and 435nm. Single channel measurements integrate the signal normally available in all imaging setup and therefore have a better signal-to-noise ratio. Resolving both the fluorescence emission spectrum and fluorescence lifetime provides the opportunity to discriminate multiple fluorophores. This instrument is intended for NAD(P)H and flavin measurements for the dynamic monitoring of cellular metabolism and optical measurements of cancerous tissue. Initial results from a study of live cells and a clinical study of human skin lesions are presented.
Owen D M, Manning H B, de Beule P, et al., 2007, Rapid hyperspectral fluorescence lifetime imaging, IMAGING, MANIPULATION AND ANALYSIS OF BIOMOLECULES, CELLS, AND TISSUES V, BIOS, Publisher: SPIE
Onfelt B, Nedvetzki S, Benninger RKP, et al., 2006, Structurally distinct membrane nanotubes between human macrophages support long-distance vesicular traffic or surfing of bacteria, JOURNAL OF IMMUNOLOGY, Vol: 177, Pages: 8476-8483, ISSN: 0022-1767
Lekadir, Karim, Elson, et al., 2006, Tissue Characterization using Dimensionality Reduction and Fluorescence Imaging, Medical Image Computing and Computer Assisted Intervention, Publisher: Medical Image Computing and Computer Assisted Intervention
Treanor B, Lanigan PMP, Kumar S, et al., 2006, Microclusters of inhibitory killer immunoglobulin like receptor signaling at natural killer cell immunological synapses, Journal of Cell Biology, Vol: 174, Pages: 153-161, ISSN: 1540-8140
We report the supramolecular organization of killer Ig–like receptor (KIR) phosphorylation using a technique applicable to imaging phosphorylation of any green fluorescent protein–tagged receptor at an intercellular contact or immune synapse. Specifically, we use fluorescence lifetime imaging (FLIM) to report Förster resonance energy transfer (FRET) between GFP-tagged KIR2DL1 and a Cy3-tagged generic anti-phosphotyrosine monoclonal antibody. Visualization of KIR phosphorylation in natural killer (NK) cells contacting target cells expressing cognate major histocompatibility complex class I proteins revealed that inhibitory signaling is spatially restricted to the immune synapse. This explains how NK cells respond appropriately when simultaneously surveying susceptible and resistant target cells. More surprising, phosphorylated KIR was confined to microclusters within the aggregate of KIR, contrary to an expected homogeneous distribution of KIR signaling across the immune synapse. Also, yellow fluorescent protein–tagged Lck, a kinase important for KIR phosphorylation, accumulated in a multifocal distribution at inhibitory synapses. Spatial confinement of receptor phosphorylation within the immune synapse may be critical to how activating and inhibitory signals are integrated in NK cells.
Zhang HX, Gu E, Jeon CW, et al., 2006, Microstripe-array InGaN light-emitting diodes with individually addressable elements, IEEE PHOTONICS TECHNOLOGY LETTERS, Vol: 18, Pages: 1681-1683, ISSN: 1041-1135
Owen DM, Lanigan PMP, Dunsby C, et al., 2006, Fluorescence lifetime imaging provides enhanced contrast when imaging the phase-sensitive dye di-4-ANEPPDHQ in model membranes and live cells, BIOPHYSICAL JOURNAL, Vol: 90, Pages: L80-L82, ISSN: 0006-3495
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