481 results found
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
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
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.
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
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
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
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 PLC epsilon contributes to chemotaxis of fibroblasts towards PDGF, JOURNAL OF CELL SCIENCE, Vol: 125, Pages: 5758-5769, ISSN: 0021-9533
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.
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
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
Xavier GDS, Mondragon A, Sun G, et al., 2012, Abnormal glucose tolerance and insulin secretion in pancreas-specific Tcf7l2-null mice, DIABETOLOGIA, Vol: 55, Pages: 2667-2676, ISSN: 0012-186X
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
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
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
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
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
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, ISSN: 1544-9173
Benati E, Bellini V, Borsari S, et al., 2011, Quantitative evaluation of healthy epidermis by means of multiphoton microscopy and fluorescence lifetime imaging microscopy, SKIN RESEARCH AND TECHNOLOGY, Vol: 17, Pages: 295-303, ISSN: 0909-752X
Talbot CB, Patalay R, Munro I, et al., 2011, Application of ultrafast gold luminescence to measuring the instrument response function for multispectral multiphoton fluorescence lifetime imaging., Opt Express, Vol: 19, Pages: 13848-13861
When performing multiphoton fluorescence lifetime imaging in multiple spectral emission channels, an instrument response function must be acquired in each channel if accurate measurements of complex fluorescence decays are to be performed. Although this can be achieved using the reference reconvolution technique, it is difficult to identify suitable fluorophores with a mono-exponential fluorescence decay across a broad emission spectrum. We present a solution to this problem by measuring the IRF using the ultrafast luminescence from gold nanorods. We show that ultrafast gold nanorod luminescence allows the IRF to be directly obtained in multiple spectral channels simultaneously across a wide spectral range. We validate this approach by presenting an analysis of multispectral autofluorescence FLIM data obtained from human skin ex vivo.
Talbot CB, Patalay R, Munro I, et al., 2011, Application of ultrafast gold luminescence to measuring the instrument response function for multispectral multiphoton fluorescence lifetime imaging, OPTICS EXPRESS, Vol: 19, Pages: 13848-13861, ISSN: 1094-4087
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
McGinty J, Stuckey DW, Soloviev VY, et al., 2011, In vivo fluorescence lifetime tomography of a FRET probe expressed in mouse, Biomedical Optics Express, Vol: 2, Pages: 1907-1917, ISSN: 2156-7085
Förster resonance energy transfer (FRET) is a powerful biological tool for reading out cell signaling processes. In vivo use of FRET is challenging because of the scattering properties of bulk tissue. By combining diffuse fluorescence tomography with fluorescence lifetime imaging (FLIM), implemented using wide-field time-gated detection of fluorescence excited by ultrashort laser pulses in a tomographic imaging system and applying inverse scattering algorithms, we can reconstruct the three dimensional spatial localization of fluorescence quantum efficiency and lifetime. We demonstrate in vivo spatial mapping of FRET between genetically expressed fluorescent proteins in live mice read out using FLIM. Following transfection by electroporation, mouse hind leg muscles were imaged in vivo and the emission of free donor (eGFP) in the presence of free acceptor (mCherry) could be clearly distinguished from the fluorescence of the donor when directly linked to the acceptor in a tandem (eGFP-mCherry) FRET construct.
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