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LIFA Widefield

作者: Kevin Peng    发布于: 2014-06-21 00:31    点击:

Fast frequency-domain FLIM microscope attachment


The Lambert Instruments Fluorescence lifetime Attachment (LIFA) is a fast frequency-domain fluorescence lifetime imaging microscopy system compatible with Leica, Nikon, Olympus and Zeiss widefield fluorescence microscopes.

You can see the LIFA at one of our Test Centers.



The LIFA - Lambert Instruments Fluorescence lifetime Attachment - is a camera-based fast fluorescence lifetime imaging microscopy system that operates in the frequency domain and is compatible with Leica, Nikon, Olympus, TILL and Zeiss fluorescence microscopes.

The well-established homodyne detection technology together with the massive parallelism of the state-of-the-art intensified camera allows near instantaneous acquisition of full field lifetime images. The widefield system includes a Multi-LED modulated light source. Its high-power LEDs can be modulated in a broad frequency range, resulting in good lifetime sensitivity and high accuracy. The components of the LIFA are the TRiCAM modulated intensified CCD camera, a dual signal generator, and the LI-FLIM software package.

The LIFA is easy to install - within the hour - and very easy to operate. The LIFA is in addition well suited for high-content screening applications. The LIFA system has been judged "easy and highly quantitative" for a.o. FLIM-FRET studies.


Hardware features

  • Fast fluorescence lifetime imaging microscopy, up to two lifetime images per second
  • Non-phototoxic illumination offered by the Multi-LED (NEW)
  • Lifetime sensitivity 0-300 ns
  • Lifetime accuracy 30 ps r.m.s.
  • High quantum efficiency with the Gen III GaAs image intensifier (Optional)

Software features

  • FRET efficiency mapping
  • Time-lapses
  • Multi-frequency acquisition and analysis for separation of multiple lifetimes
  • Polar (Phasor) plot inspection and separation of multiple lifetimes
  • Easy integration into specialized image analysis pipelines through ActiveX
  • Export to Metamorph, ImageJ, Matlab
  • Images can be exported as BMP, TIFF
  • Statistics and histograms can be exported to MS Excel


Lambert Instruments provides the following LIFA models, which cover a wide range in spectral sensitivity and in lifetime sensitivity. Each model is based on a specific high-resolution Gen II or Gen III image intensifier, see also our fluorescence lifetime imaging microscopy properties of modulated intensifiers document.

Model

Photocathode

Modulation frequency

Lifetime sensitivity

LIFA Gen II S20 S20 1-120 MHz 0-300 ns
LIFA Gen II S25 S25 1-120 MHz 0-300 ns
LIFA Gen III GaAs GaAs 1-120 MHz 0-300 ns
LIFA X Gen II S20 S20 0-100 kHz, 1-120 MHz 0 ns - 1 ms
LIFA X Gen II S25 S25 0-100 kHz, 1-120 MHz 0 ns - 1 ms
LIFA X Gen III GaAs GaAs 0-100 kHz, 1-120 MHz 0 ns - 1 ms
LIFA P Gen II S20 S20 0-100 kHz 100 ns - 1 ms
LIFA P Gen II S25 S25 0-100 kHz 100 ns - 1 ms
LIFA P Gen III GaAs GaAs 0-100 kHz 100 ns - 1 ms
LIFA P Gen III GaAsP GaAsP 0-100 kHz 100 ns - 1 ms

 

Photocathode spectral sensitivity

Spectral sensivity as a function of wavelength for each of the photocathodes is shown below.

Spectral sensitivity of photocathodes

For more information, please refer to our technologies page on the operating principle of image intensifiers for lifetime imaging.

Options

 

Support for automated microscopes

Nikon Ti Eclipse.

Support for XYZ stages


The LIFA has the following applications:

  • Molecular interactions
  • Protein conformation
  • Biosensors
  • Oxygen concentration imaging in cells and tissue
  • NADH/FAD fluorescence dynamics
  • Viscosity imaging
  • Membrane dynamics
  • Membrane trafficking
  • LED inspection
  • Crude oil characterisation





Selected LIFA Publications

Abankwa, D., et al., The efficacy of Raf kinase recruitment to the GTPase H-ras depends on H-ras membrane conformer-specific nanoclustering, J Biol Chem. (2014)

Goedhart J. et al., Structure-guided evolution of cyan fluorescent proteins towards a quantum yield of 93%, Nature Communications (2012), 3:751

José Pena, E., et al.,Citrus psorosis and Mirafiori lettuce big-vein ophiovirus coat proteins localize to the cytoplasm and self interact in vivo, Virus Research (2012)

Byrne, R.D., et al., Dynamics of PLCγ and Src Family Kinase 1 Interactions during Nuclear Envelope Formation Revealed by FRET-FLIM, PLoS One. (2012), 7(7): e40669

Zhang, H., et al., Regulation of AMPA receptor surface trafficking and synaptic plasticity by a cognitive enhancer and antidepressant molecule, Molecular Psychiatry advance online publication 26 June 2012

Pereira, A.M., et al., Integrin-Dependent Activation of the JNK Signaling Pathway by Mechanical Stress, PLoS One. (2011), 6(12): e26182

Praus, A., et al., Cellular uptake of modified oligonucleotides enhanced by porphyrins studied by time-resolved microspectrofluorimetry and fluorescence imaging techniques, Journal of Molecular Structure 993 (2011) 316-318

Zhao Q, Young IT, de Jong JG., Photon budget analysis for fluorescence lifetime imaging microscopy, J Biomed Opt. 2011 Aug;16(8):086007

Klarenbeek, JB, A mTurquoise-Based cAMP Sensor for Both FLIM and Ratiometric Read-Out Has Improved Dynamic Range, PLoS One. 2011 Apr 29;6(4):e19170

Dragavon J, et al., Fluorescence lifetime imaging to quantify sub-cellular oxygen measurements in live macrophage during bacterial invasion, Proc. SPIE 7910, 791019 (2011); doi:10.1117/12.875430

Svensson FR, et al., Ruthenium(II) Complex Enantiomers as Cellular Probes for Diastereomeric Interactions in Confocal and Fluorescence Lifetime Imaging Microscopy, J. Phys. Chem. Lett. (2011) 2:397–401

Huntosova V., et al., Interaction dynamics of hypericin with low-density lipoproteins and U87-MG cells, International Journal of Pharmaceutics 389 (2010) 32-40

Vos MJ, et al., HSPB7 is the most potent polyQ aggregation suppressor within the HSPB family of molecular chaperones, Human Molecular Genetics (2010) 19(23):4677-93 Kozer, N., et al., Creation and biophysical characterization of a high-affinity, monomeric EGF receptor ectodomain using fluorescent proteins, Biochemistry (2010) 49(35):7459-66

Hageman, J., et al., A DNAJB Chaperone Subfamily with HDAC-dependent Activities Suppresses Toxic Protein Aggregation. Molecular Cell (2010) 37(3):355-69

Abankwa D, et al., Ras membrane orientation and nanodomain localization generate isoform diversity. Proc Natl Acad Sci (2010) 107(3):1130-5

Bastiani M, et al., MURC/Cavin-4 and cavin family members form tissue-specific caveolar complexes. J Cell Biol (2009) 185(7):1259-73

Aymeric Leray A., et al., Optimized protocol of a frequency domain fluorescence lifetime imaging microscope for FRET measurements, Microscopy Research and Technique (2009) 72(5) 371-379

Hafrén J., et al., Fluorescence lifetime imaging microscopy study of wood fibers. J Wood Sci (2009) 55(3) 236-239

Schlachter S., et al., mhFLIM: Resolution of heterogeneous fluorescence decays in widefield lifetime microscopy. Optics Express (2009) 17(3):1557-70

Valdembri D, et al., Neuropilin-1/GIPC1 signaling regulates alpha5beta1 integrin traffic and function in endothelial cells. PLoS Biol. (2009) 27:7(1):e25

Gadella TW Jr., FRET and FLIM techniques, 33. Imprint: Elsevier, ISBN-13: 978-0-08-054958-3. (2008) 560 pages

Langel FD, et al., Multiple protein domains mediate interaction between Bcl10 and Malt1, J. Biol. Chem., (2008) 283(47):32419-31

Clayton AH. , The polarized AB plot for the frequency-domain analysis and representation of fluorophore rotation and resonance energy homotransfer. J Microsc. (2008) 232(2):306-12

Clayton AH, et al., Predominance of activated EGFR higher-order oligomers on the cell surface. Growth Factors (2008) 20:1

Plowman et al., Electrostatic Interactions Positively Regulate K-Ras Nanocluster Formation and Function. Molecular and Cellular Biology (2008) 4377–4385

Belanis L, et al., Galectin-1 Is a Novel Structural Component and a Major Regulator of H-Ras Nanoclusters. Molecular Biology of the Cell (2008) 19:1404–1414

Van Manen HJ, Refractive index sensing of green fluorescent proteins in living cells using fluorescence lifetime imaging microscopy. Biophys J. (2008) 94(8):L67-9

Van der Krogt GNM, et al., A Comparison of Donor-Acceptor Pairs for Genetically Encoded FRET Sensors: Application to the Epac cAMP Sensor as an Example, PLoS ONE, (2008) 3(4):e1916

Dai X, et al., Fluorescence intensity and lifetime imaging of free and micellar-encapsulated doxorubicin in living cells. Nanomedicine. (2008) 4(1):49-56

Elder A, et al., Theoretical investigation of the photon efficiency in frequency-domain fluorescence lifetime imaging microscopy. J Opt Soc Am A Opt Image Sci Vis. (2008) 25(2):452-62.

Berdiev BK, et al., Molecular proximity of CFTR and ENaC assessed by fluorescence resonance energy transfer, J. Biol. Chem., (2007) 282(50):36481-88

Domingo B, et al., Imaging FRET standards by steady-state fluorescence and lifetime methods, Microsc Res Tech. (2007) 70(12):1010-21

Matthews SM, et al., Quantitative kinetic analysis in a microfluidic device using frequency-domain fluorescence lifetime imaging, Anal Chem. (2007) 79(11):4101-9

Tian T, et al., Plasma membrane nanoswitches generate high-fidelity Ras signal transduction, Nat Cell Biol. (2007) 9(8):905-14

Clayton AHA, et al., Unligated epidermal growth factor receptor forms higher order oligomers within microclusters on A431 cells that are sensitive to tyrosine kinase inhibitor binding. Biochemistry (2007) 46(15):4589-97

Elder AD, et al, Calibration of a wide-field frequency-domain fluorescence lifetime microscopy system using light emitting diodes as light sources, Journal of Microscopy (2006) 224(Pt2):166-80

Elder AD, et al., Application of frequency-domain Fluorescence Lifetime Imaging Microscopy as a quantitative analytical tool for microfluidic devices, Optics Express (2006) 14:5456-5467

Dai X, et al., A spectroscopic study of the self-association and inter-molecular aggregation behaviour of pH-responsive poly(l-lysine iso-phtalamide), Polymer (2006) 47(8):2689-2698

Clayton AHA, et al, Ligand-induced dimer-tetramer transition during the activation of the cell surface epidemal growth factor receptor-a multidimensional microscopy analysis, Journal of Biological Chemistry (2005) 280(34):30392-30399

Van Rheenen J, et al.,PIP2 signaling in lipid domains: a critical re-evaluation, The EMBO Journal (2005) 24(9):1664–1673

Hanley QS and Clayton AHA, AB-plot assisted determination of fluorophore mixtures in a fluorescence lifetime microscope using spectra or quenchers, Journal of Microscopy (2005) 218(1):62-7

Zwart W, et al., Spatial separation of HLA-DM/HLA-DR interactions within MIIC and phagosome-induced immune escape, Immunity (2005), 22(2):221-233

Ponsioen B, et al., Detecting cAMP-induced Epac activation by fluorescence resonance energy transfer: Epac as a novel cAMP indicator, EMBO reports (2004) 5(12):1176–1180

May M, An easy upgrade to fluorescence lifetime imaging, BioPhotonics International (2004) 20-21

Stoop KWJ, et al., Measuring FRET in living cells with FLIM, 8th Chinese Peptide Symposium, Kunming China, (2004) July 3-6

Van Geest LK and Stoop KWJ, FLIM on a wide field fluorescence microscope, Letters in Peptide Science (2003) 10(5-6):501-510

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