Publications
300 results found
Burgin E, Salehi-Reyhani A, Barclay M, et al., 2014, Absolute quantification of protein copy number using a single-molecule-sensitive microarray, ANALYST, Vol: 139, Pages: 3235-3244, ISSN: 0003-2654
Lenz MO, Sinclair HG, Savell A, et al., 2014, 3-D stimulated emission depletion microscopy with programmable aberration correction, Journal of Biophotonics, Vol: 7, Pages: 29-36, ISSN: 1864-063X
We present a stimulated emission depletion (STED) microscope that provides 3‐D super resolution by simultaneous depletion using beams with both a helical phase profile for enhanced lateral resolution and an annular phase profile to enhance axial resolution. The 3‐D depletion point spread function is realised using a single spatial light modulator that can also be programmed to compensate for aberrations in the microscope and the sample. We apply it to demonstrate the first 3‐D super‐resolved imaging of an immunological synapse between a Natural Killer cell and its target cell.
Sonnefraud Y, Sivan Y, Sinclair HG, et al., 2014, Nanoparticle-assisted STED, theory, and experimental demonstration, Conference on Nanoimaging and Nanospectroscopy II, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Soltan A, Zhao H, Chaudet L, et al., 2014, An 8100 pixel optoelectronic array for optogenetic retinal prosthesis, IEEE Biomedical Circuits and Systems Conference (BioCAS), Publisher: IEEE, Pages: 352-355, ISSN: 2163-4025
- Author Web Link
- Cite
- Citations: 7
Schrems A, Phillips J, Casey D, et al., 2014, The grab-and-drop protocol: a novel strategy for membrane protein isolation and reconstitution from single cells, ANALYST, Vol: 139, Pages: 3296-3304, ISSN: 0003-2654
- Author Web Link
- Cite
- Citations: 8
Salehi-Reyhani A, Sharma S, Burgin E, et al., 2014, Scaling advantages and constraints in miniaturized capture assays for single cell protein analysis (vol 13, pg 2066, 2013), LAB ON A CHIP, Vol: 14, Pages: 3430-3430, ISSN: 1473-0197
Manivannan N, Neil MAA, Balachandran W, 2013, Optical alignment of pixelated 4<i>f</i> optical system using multiplexed filter, APPLIED OPTICS, Vol: 52, Pages: 7812-7820, ISSN: 1559-128X
- Author Web Link
- Cite
- Citations: 2
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.
Clegg JH, Neil MAA, 2013, Double pass, common path method for arbitrary polarization control using a ferroelectric liquid crystal spatial light modulator, OPTICS LETTERS, Vol: 38, Pages: 1043-1045, ISSN: 0146-9592
- Author Web Link
- Cite
- Citations: 26
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).
Neil MAA, 2013, Optical manipulation of chemically functionalised emulsions and suspensions for single cell analysis
Optical traps are used to manipulate chemically modified micro-emulsions and particles in micro-suspension for single cell manipulation and analysis. Material extraction from and introduction to cells is possible by modifying the droplet or particle coating. © 2013 The Optical Society (OSA).
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).
Hargreaves AL, Kirby AK, Bain CD, et al., 2013, An optical platform for the production, trapping, manipulation and visualization of ultra-low interfacial tension emulsion droplets, Conference on Optical Trapping and Optical Micromanipulation X, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
- Author Web Link
- Cite
- Citations: 1
Fafchamps LJ, Neil MAA, Juskaitis R, 2013, GPU-based Image Registration in Aperture Correlation Microscopy, and Reflection Mode Correlation Microscopy, Conference on Three-Dimensional and Multidimensional Microscopy - Image Acquisition and Processing XX, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
Salehi-Reyhani A, Sharma S, Burgin E, et al., 2013, Scaling Advantages and Constraints in Miniaturized Capture Assays for Single Cell Protein Analysis, Lab on A Chip, Vol: 13, Pages: 2066-2074, ISSN: 1473-0197
Measuring protein expression in single cells is the basis of single cell proteomics. The sensitivity and dynamic range of a single cell immunoassay should ideally be such that proteins that are expressed between 1 – 106 copies per cell can be detected and counted. We have investigated the effect of miniaturizing antibody microarrays by reducing capture spot sizes from 100 μm to 15 μm using dip pen nanolithography. We demonstrate that protocols developed for printing and passivating antibody capture spots using conventional pin based contact printing can be directly transferred to dip-pen lithography whilst retaining the capture activity per unit area. Using a simple kinetic model, we highlight how the limit of detection and dynamic range of a sandwich immunoassay, respectively, increase and decrease when spot size is reduced. However, we show that reducing spot size is more effective than increasing assay chamber volume when seeking to multiplex such a microfluidic immunoassay. Although, we make particular reference to single cell microfluidic immunoassays, the topics discussed here are applicable to capture assays in general.
Chaudet L, Neil M, Degenaar P, et al., 2013, Development of Optics with Micro-LED Arrays for Improved Opto-electronic Neural Stimulation, Conference on Optogenetics - Optical Methods for Cellular Control, Publisher: SPIE-INT SOC OPTICAL ENGINEERING, ISSN: 0277-786X
- Author Web Link
- Cite
- Citations: 8
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.
Wylie D, Casey D, Phillips J, et al., 2012, Novel nanotechnologies for multiple spatially and temporally resolved live single cell membrane sampling and analysis, FREE RADICAL BIOLOGY AND MEDICINE, Vol: 53, Pages: S127-S128, ISSN: 0891-5849
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, Talbot C, Alexandrov Y, et al., 2012, Basal cell carcinoma evaluation and tumour margin assessment using multiphoton tomography and fluorescence lifetime imaging, 92nd Annual Meeting of the British-Association-of-Dermatologists, Publisher: WILEY-BLACKWELL, Pages: 78-79, ISSN: 0007-0963
- Author Web Link
- Cite
- Citations: 1
Manivannan N, Neil MAA, Balachandran W, 2012, Optical correlator-neural networks hybrid system for automatic angle measurement of two-dimensional objects, OPTICAL ENGINEERING, Vol: 51, ISSN: 0091-3286
- Author Web Link
- Cite
- Citations: 1
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
- Author Web Link
- Cite
- Citations: 30
Neil MAA, Manivannan N, 2012, Analysis of coherent optical filter and correlator systems using pixellated spatial light modulators, IET OPTOELECTRONICS, Vol: 6, Pages: 43-51, ISSN: 1751-8768
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
- Author Web Link
- Cite
- Citations: 18
Lanigan PMP, Munro I, Grace EJ, et al., 2012, Dynamical hologram generation for high speed optical trapping of smart droplet microtools, Biomedical Optics Express, Vol: 3, Pages: 1609-1609
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.
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
- Author Web Link
- Cite
- Citations: 27
This data is extracted from the Web of Science and reproduced under a licence from Thomson Reuters. You may not copy or re-distribute this data in whole or in part without the written consent of the Science business of Thomson Reuters.