fluorescence recovery after photobleaching

Summary

Summary: A method used to study the lateral movement of MEMBRANE PROTEINS and LIPIDS. A small area of a cell membrane is bleached by laser light and the amount of time necessary for unbleached fluorescent marker-tagged proteins to diffuse back into the bleached site is a measurement of the cell membrane's fluidity. The diffusion coefficient of a protein or lipid in the membrane can be calculated from the data. (From Segen, Current Med Talk, 1995).

Top Publications

  1. ncbi Measurement of dynamic protein binding to chromatin in vivo, using photobleaching microscopy
    Robert D Phair
    BioInformatics Services, Rockville, Maryland 20854, USA
    Methods Enzymol 375:393-414. 2004
  2. ncbi Live-cell imaging reveals a stable cohesin-chromatin interaction after but not before DNA replication
    Daniel Gerlich
    Gene Expression Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany
    Curr Biol 16:1571-8. 2006
  3. ncbi Dynamic changes in the epigenomic state and nuclear organization of differentiating mouse embryonic stem cells
    Satoru Kobayakawa
    Graduate School of Life and Environmental Sciences, University of Tsukuba, 1 1 1 Ten noudai, Tsukuba, Ibaraki 305 8572, Japan
    Genes Cells 12:447-60. 2007
  4. pmc Arp2/3 complex interactions and actin network turnover in lamellipodia
    Frank P L Lai
    Cytoskeleton Dynamics Group, Helmholtz Centre for Infection Research, Braunschweig, Germany
    EMBO J 27:982-92. 2008
  5. pmc Analysis of binding reactions by fluorescence recovery after photobleaching
    Brian L Sprague
    Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
    Biophys J 86:3473-95. 2004
  6. ncbi Development and use of fluorescent protein markers in living cells
    Jennifer Lippincott-Schwartz
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Science 300:87-91. 2003
  7. ncbi Myosin-II-dependent localization and dynamics of F-actin during cytokinesis
    Kausalya Murthy
    Department of Biology and Program in Molecular and Cellular Biology, University of Massachusetts, Amherst, 01003, USA
    Curr Biol 15:724-31. 2005
  8. pmc Vesicles carry most exocyst subunits to exocytic sites marked by the remaining two subunits, Sec3p and Exo70p
    Charles Boyd
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
    J Cell Biol 167:889-901. 2004
  9. pmc Mobility of cytoplasmic, membrane, and DNA-binding proteins in Escherichia coli
    Mohit Kumar
    Zentrum fur Molekulare Biologie der Universitat Heidelberg, DKFZ ZMBH Alliance, Heidelberg, Germany
    Biophys J 98:552-9. 2010
  10. pmc Rapid glucocorticoid receptor exchange at a promoter is coupled to transcription and regulated by chaperones and proteasomes
    Diana A Stavreva
    Laboratory of Receptor Biology and Gene Expression, Center for Cancer Research, National Cancer Institute Light Imaging Facility, National Institute for Neurological Disorders and Stroke, Bethesda, Maryland 20892, USA
    Mol Cell Biol 24:2682-97. 2004

Detail Information

Publications312 found, 100 shown here

  1. ncbi Measurement of dynamic protein binding to chromatin in vivo, using photobleaching microscopy
    Robert D Phair
    BioInformatics Services, Rockville, Maryland 20854, USA
    Methods Enzymol 375:393-414. 2004
    ..These methods should prove useful in the further in vivo investigation of the molecular mechanisms involved in genome organization and expression...
  2. ncbi Live-cell imaging reveals a stable cohesin-chromatin interaction after but not before DNA replication
    Daniel Gerlich
    Gene Expression Unit, European Molecular Biology Laboratory, 69117 Heidelberg, Germany
    Curr Biol 16:1571-8. 2006
    ....
  3. ncbi Dynamic changes in the epigenomic state and nuclear organization of differentiating mouse embryonic stem cells
    Satoru Kobayakawa
    Graduate School of Life and Environmental Sciences, University of Tsukuba, 1 1 1 Ten noudai, Tsukuba, Ibaraki 305 8572, Japan
    Genes Cells 12:447-60. 2007
    ..Thus, this experimental system should facilitate studies focusing on relationships between nuclear organization, epigenetic status and cell differentiation...
  4. pmc Arp2/3 complex interactions and actin network turnover in lamellipodia
    Frank P L Lai
    Cytoskeleton Dynamics Group, Helmholtz Centre for Infection Research, Braunschweig, Germany
    EMBO J 27:982-92. 2008
    ..Here, we combine fluorescence recovery after photobleaching (FRAP) of different lamellipodial components with a new method of data analysis to shed light on ..
  5. pmc Analysis of binding reactions by fluorescence recovery after photobleaching
    Brian L Sprague
    Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
    Biophys J 86:3473-95. 2004
    b>Fluorescence recovery after photobleaching (FRAP) is now widely used to investigate binding interactions in live cells...
  6. ncbi Development and use of fluorescent protein markers in living cells
    Jennifer Lippincott-Schwartz
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892, USA
    Science 300:87-91. 2003
    ....
  7. ncbi Myosin-II-dependent localization and dynamics of F-actin during cytokinesis
    Kausalya Murthy
    Department of Biology and Program in Molecular and Cellular Biology, University of Massachusetts, Amherst, 01003, USA
    Curr Biol 15:724-31. 2005
    ..The mechanism(s) that contribute to the spatially and temporally regulated assembly and disassembly of the CR at the cell equator are poorly understood...
  8. pmc Vesicles carry most exocyst subunits to exocytic sites marked by the remaining two subunits, Sec3p and Exo70p
    Charles Boyd
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
    J Cell Biol 167:889-901. 2004
    ..Assembly of the exocyst occurs when the first subset of subunits, delivered on vesicles, joins Sec3p and Exo70p on the plasma membrane. Exocyst assembly serves to both target and tether vesicles to sites of exocytosis...
  9. pmc Mobility of cytoplasmic, membrane, and DNA-binding proteins in Escherichia coli
    Mohit Kumar
    Zentrum fur Molekulare Biologie der Universitat Heidelberg, DKFZ ZMBH Alliance, Heidelberg, Germany
    Biophys J 98:552-9. 2010
    ..Here we combined fluorescence recovery after photobleaching with numerical modeling to analyze mobility of a set of fluorescent protein fusions in the ..
  10. pmc Rapid glucocorticoid receptor exchange at a promoter is coupled to transcription and regulated by chaperones and proteasomes
    Diana A Stavreva
    Laboratory of Receptor Biology and Gene Expression, Center for Cancer Research, National Cancer Institute Light Imaging Facility, National Institute for Neurological Disorders and Stroke, Bethesda, Maryland 20892, USA
    Mol Cell Biol 24:2682-97. 2004
    ..Moreover, our results reveal that longer GR residence times are consistently associated with greater transcriptional output, suggesting a new paradigm in which the rate of rapid exchange provides a means to tune transcriptional levels...
  11. pmc Stress fibers are generated by two distinct actin assembly mechanisms in motile cells
    Pirta Hotulainen
    Institute of Biotechnology, University of Helsinki, Helsinki FI 00014, Finland
    J Cell Biol 173:383-94. 2006
    ..b>Fluorescence recovery after photobleaching analysis revealed that actin filament cross-linking in stress fibers is highly dynamic, ..
  12. pmc Direct measurement of association and dissociation rates of DNA binding in live cells by fluorescence correlation spectroscopy
    Ariel Michelman-Ribeiro
    Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
    Biophys J 97:337-46. 2009
    ..We find that our FCS estimates are quantitatively consistent with our fluorescence recovery after photobleaching (FRAP) measurements on the same VBP domains...
  13. ncbi Dynamics of single mRNPs in nuclei of living cells
    Yaron Shav-Tal
    Departments of Anatomy and Structural Biology and Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA
    Science 304:1797-800. 2004
    ..This observation resolves a controversy, showing that the energetic requirements of nuclear mRNP trafficking are consistent with a diffusional model...
  14. pmc Dynamics of putative raft-associated proteins at the cell surface
    Anne K Kenworthy
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bldg 18T, Rm 101, 18 Library Dr, Bethesda, MD 20895, USA
    J Cell Biol 165:735-46. 2004
    ..The properties of these domains in vivo are unclear. Here, we use fluorescence recovery after photobleaching to test if raft association affects a protein's ability to laterally diffuse large distances ..
  15. pmc Ligand-specific dynamics of the progesterone receptor in living cells and during chromatin remodeling in vitro
    Geetha V Rayasam
    Laboratory of Receptor Biology and Gene Expression, Building 41, B602, National Cancer Institute, National Institutes of Health, 41 Library Dr, Bethesda, MD 20892 5055, USA
    Mol Cell Biol 25:2406-18. 2005
    ..of the interaction of PR with a natural target promoter in living cells through the use of fluorescence recovery after photobleaching (FRAP) analysis and also have characterized the dynamics of the interaction of PR with the mouse ..
  16. pmc Evidence for a common mode of transcription factor interaction with chromatin as revealed by improved quantitative fluorescence recovery after photobleaching
    Florian Mueller
    Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, Bethesda, Maryland 20892, USA
    Biophys J 94:3323-39. 2008
    ..transcription factors with chromatin have been investigated recently using quantitative fluorescence recovery after photobleaching (FRAP)...
  17. ncbi Dynamic nuclear actin assembly by Arp2/3 complex and a baculovirus WASP-like protein
    Erin D Goley
    Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA
    Science 314:464-7. 2006
    ..Recompartmentalizing dynamic host actin may represent a conserved mode of pathogenesis and reflect viral manipulation of normal functions of nuclear actin...
  18. ncbi Polyglutamine protein aggregates are dynamic
    Soojin Kim
    Department of Biochemistry, Molecular Biology and Cell Biology, Rice Institute for Biomedical Research, Northwestern University, Evanston, IL 60208, USA
    Nat Cell Biol 4:826-31. 2002
    ..These studies provide new insights into the composite organization and formation of protein aggregates and show that molecular chaperones are not sequestered into aggregates, but are instead transiently associated...
  19. pmc Human ISWI chromatin-remodeling complexes sample nucleosomes via transient binding reactions and become immobilized at active sites
    Fabian Erdel
    Research Group Genome Organization and Function, Deutsches Krebsforschungszentrum and BIOQUANT, 69120 Heidelberg, Germany
    Proc Natl Acad Sci U S A 107:19873-8. 2010
    ..Due to the relatively high intranuclear remodeler concentrations cellular response times for repositioning a given nucleosome were calculated to be in the range of tens of seconds to minutes...
  20. ncbi Dynamic interaction between BAF and emerin revealed by FRAP, FLIP, and FRET analyses in living HeLa cells
    Takeshi Shimi
    CREST of JST, Kansai Advanced Research Center, CRL, Kobe 651 2492, Japan
    J Struct Biol 147:31-41. 2004
    ..Using fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP), we compared the mobility of BAF to its ..
  21. pmc FRAP and kinetic modeling in the analysis of nuclear protein dynamics: what do we really know?
    Florian Mueller
    Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, Bethesda, MD 20892, USA
    Curr Opin Cell Biol 22:403-11. 2010
    ..in live cells has been analyzed by kinetic modeling procedures applied to experimental data from fluorescence recovery after photobleaching (FRAP)...
  22. pmc Protein diffusion in mammalian cell cytoplasm
    Thomas Kuhn
    Nanoscience Center, Department of Physics, University of Jyvaskyla, Jyvaskyla, Finland
    PLoS ONE 6:e22962. 2011
    ..The cytosol results were found to be in very good agreement with those by FCS...
  23. pmc Cross-validating FRAP and FCS to quantify the impact of photobleaching on in vivo binding estimates
    Timothy J Stasevich
    National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
    Biophys J 99:3093-101. 2010
    ..To address this uncertainty, we compare fluorescence recovery after photobleaching (FRAP) and fluorescence correlation spectroscopy (FCS) measurements of the binding kinetics of a ..
  24. pmc Dosage-sensitive regulation of cohesin chromosome binding and dynamics by Nipped-B, Pds5, and Wapl
    Maria Gause
    Edward A Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO 63104, USA
    Mol Cell Biol 30:4940-51. 2010
    ..We addressed this question by in vivo fluorescence recovery after photobleaching (FRAP) with Drosophila salivary glands...
  25. ncbi Effects of interphase and mitotic phosphorylation on the mobility and location of nucleolar protein B23
    Sandeep S Negi
    Department of Biochemistry, University of Mississippi Medical Center, Jackson, MS 39216, USA
    J Cell Sci 119:3676-85. 2006
    ..results in slower recovery of the mutant compared with the wild-type protein as measured by fluorescence recovery after photobleaching (FRAP)...
  26. ncbi The interaction between Stargazin and PSD-95 regulates AMPA receptor surface trafficking
    Cecile Bats
    Physiologie Cellulaire de la Synapse, UMR 5091 CNRS Institut François Magendie, Universite Bordeaux, Bordeaux 33077, France
    Neuron 53:719-34. 2007
    ..These results propose a model in which the Stargazin-PSD-95 interaction plays a key role to trap and transiently stabilize diffusing AMPARs in the postsynaptic density...
  27. pmc Compartmentalization of androgen receptor protein-protein interactions in living cells
    Martin E van Royen
    Department of Pathology, Josephine Nefkens Institute, Erasmus MC, 3000 CA Rotterdam, Netherlands
    J Cell Biol 177:63-72. 2007
    ..N/C interactions are largely lost when AR transiently binds to DNA, predominantly in foci partly overlapping transcription sites. AR coregulator interactions occur preferentially when ARs are bound to DNA...
  28. pmc Magnitude and direction of vesicle dynamics in growing pollen tubes using spatiotemporal image correlation spectroscopy and fluorescence recovery after photobleaching
    Jérôme Bove
    Departement de sciences biologiques, Universite de Montreal, Montreal, Quebec, Canada H1X 2B2
    Plant Physiol 147:1646-58. 2008
    ..b>Fluorescence recovery after photobleaching confirmed vesicle accumulation in the shoulder of the apex, and it revealed that the extreme ..
  29. ncbi Binding of ATP to UAP56 is necessary for mRNA export
    Krishna P Kota
    Department of Cell Biology S7 214, University of Massachusetts Medical School, Worcester, MA 01655, USA
    J Cell Sci 121:1526-37. 2008
    ..This was confirmed using a point mutant of UAP56 that did not bind ATP. Point mutation of UAP56 to eliminate ATP binding did not affect RNA splicing, but strongly inhibited the export of mRNA to the cytoplasm...
  30. ncbi Membrane mobility and microdomain association of the dopamine transporter studied with fluorescence correlation spectroscopy and fluorescence recovery after photobleaching
    Erika M Adkins
    Molecular Neuropharmacology Group, Department of Neuroscience and Pharmacology and Center for Pharmacogenomics, The Panum Institute, University of Copenhagen, DK 2200 Copenhagen, Denmark
    Biochemistry 46:10484-97. 2007
    ..dopamine transporter (DAT), we employed FCS (fluorescence correlation spectroscopy) and FRAP (fluorescence recovery after photobleaching)...
  31. ncbi Mobility of late endosomal and lysosomal markers on phagosomes analyzed by fluorescence recovery after photobleaching
    Keiko Sugaya
    Department of Health Science, Hamamatsu University School of Medicine, 1 20 1 Handa yama, Higashi ku, Hamamatsu 431 3192, Japan
    Biochem Biophys Res Commun 410:371-5. 2011
    ..We investigated the mobility of Rab7 and LAMP1 on the phagosomes in macrophages by fluorescence recovery after photobleaching (FRAP) analysis...
  32. ncbi The biochemistry of RNA metabolism studied in situ
    Jeffrey A Nickerson
    Department of Cell Biology and Cancer Center, University of Massachusetts Medical School, Worcester, Massachusetts 01655, USA
    RNA Biol 6:25-30. 2009
    ..The complimentary technique of fluorescence recovery after photobleaching (FRAP) measures the rates of binding and unbinding of those molecules in their complexes...
  33. ncbi Quantification of protein-protein and protein-DNA interactions in vivo, using fluorescence recovery after photobleaching
    Gustavo Carrero
    Department of Mathematical and Statistical Sciences, Faculty of Science, University of Alberta, Edmonton, Alberta T6G 2E1, Canada
    Methods Enzymol 375:415-42. 2004
  34. pmc Role of fascin in filopodial protrusion
    Danijela Vignjevic
    Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA
    J Cell Biol 174:863-75. 2006
    ..b>Fluorescence recovery after photobleaching of GFP-tagged wild-type and S39A fascin showed that dephosphorylated fascin underwent rapid ..
  35. pmc Fluorescent recovery after photobleaching (FRAP) analysis of nuclear export rates identifies intrinsic features of nucleocytoplasmic transport
    Francesco Cardarelli
    Center for Nanotechnology Innovation NEST, Istituto Italiano di Tecnologia, Piazza San Silvestro 12 56127 Pisa, Italy
    J Biol Chem 287:5554-61. 2012
    ..These findings suggest differential gating at the NPC level...
  36. ncbi A dynamic view of cellular processes by in vivo fluorescence auto- and cross-correlation spectroscopy
    Kirsten Bacia
    Experimental Biophysics Group, Max Planck Institut fur biophysikalische Chemie, Am Fassberg 11, D 37077, Gottingen, Germany
    Methods 29:74-85. 2003
    ..Practical implications of nonideal conditions including incomplete focus overlap and spectral cross-talk are considered. Recent examples of both auto- and cross-correlation applications demonstrate the potential of FCS for cell biology...
  37. ncbi Accurate quantification of diffusion and binding kinetics of non-integral membrane proteins by FRAP
    Ronen Berkovich
    School of Chemistry, Raymond and Beverley Sackler Faculty of Exact Sciences, Tel Aviv University, Tel Aviv 69978, Israel
    Traffic 12:1648-57. 2011
    ..Despite a fair number of approximate fitting functions for analyzing fluorescence recovery after photobleaching (FRAP) data, no approach was able to cope with the full diffusion-exchange problem...
  38. ncbi Methods for measuring rates of protein binding to insoluble scaffolds in living cells: histone H1-chromatin interactions
    Tanmay Lele
    Department of Pathology, Children s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA
    J Cell Biochem 99:1334-42. 2006
    ..k(ON) and dissociation rate constant k(OFF) to be determined based on data obtained from fluorescence recovery after photobleaching (FRAP) analysis...
  39. pmc Dissecting the binding mechanism of the linker histone in live cells: an integrated FRAP analysis
    Timothy J Stasevich
    Laboratory of Receptor Biology and Gene Expression, National Cancer Institute, US National Institutes of Health, Bethesda, MD 20892 5055, USA
    EMBO J 29:1225-34. 2010
    ..Using a novel fluorescence recovery after photobleaching (FRAP) procedure, we have measured the affinities of these regions individually, in pairs, and ..
  40. pmc Endocytosis is required for E-cadherin redistribution at mature adherens junctions
    Simon de Beco
    Centre de Recherche, Institut Curie, F 75248 Paris, France
    Proc Natl Acad Sci U S A 106:7010-5. 2009
    ..the dynamics of E-cadherin were quantitatively analyzed by a new approach combining 2-photon fluorescence recovery after photobleaching (FRAP) and fast 3D wide-field fluorescence microscopy...
  41. pmc A quantitative approach to analyze binding diffusion kinetics by confocal FRAP
    Minchul Kang
    Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA
    Biophys J 99:2737-47. 2010
    ..We validate this method by measuring kinetic rate constants for the CAAX-mediated binding of Ras to membranes of the endoplasmic reticulum, obtaining binding constants of k(on) ∼ 255/s and k(off) ∼ 31/s...
  42. ncbi How to quantify protein diffusion in the bacterial membrane
    Siet M J L van den Wildenberg
    Department of Physics and Astronomy, VU University, De Boelelaan 1081, 1081HV Amsterdam, The Netherlands
    Biopolymers 95:312-21. 2011
    ..using various experimental techniques, including fluorescence correlation spectroscopy (FCS), fluorescence recovery after photobleaching (FRAP), and particle tracking using single-molecule fluorescence (SMF) microscopy...
  43. pmc Visualizing histone modifications in living cells: spatiotemporal dynamics of H3 phosphorylation during interphase
    Yoko Hayashi-Takanaka
    Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565 0871, Japan
    J Cell Biol 187:781-90. 2009
    ..Visualizing histone modifications in living cells will facilitate future epigenetic and cell regulation studies...
  44. pmc Temporal changes in microvessel leakiness during wound healing discriminated by in vivo fluorescence recovery after photobleaching
    Maria J C Machado
    Centre for Molecular Biosciences, University of Ulster, Cromore Road, Coleraine, Co Londonderry, UK
    J Physiol 589:4681-96. 2011
    ..vessels, angiogenic plexus and blind-ended vessels (BEVs)) was quantified using in vivo fluorescence recovery after photobleaching (FRAP) and linear regression analysis of recovery profiles...
  45. pmc The transcriptional cycle of HIV-1 in real-time and live cells
    Stephanie Boireau
    Institute of Molecular Genetics of Montpellier, Unite Mixte de Recherche 5535, Centre National de la Recherche Scientifique, 34293 Montpellier, France
    J Cell Biol 179:291-304. 2007
    ..The strengths and limitations of this kinetic approach to analyze mRNA biogenesis in living cells are discussed...
  46. pmc The human deafness-associated connexin 30 T5M mutation causes mild hearing loss and reduces biochemical coupling among cochlear non-sensory cells in knock-in mice
    Melanie Schütz
    Institut fuer Genetik, Rheinische Friedrich Wilhelms Universitaet Bonn, Roemerstrasse 164, D 53117 Bonn, Germany
    Hum Mol Genet 19:4759-73. 2010
    ..Our findings link hearing loss to decreased biochemical coupling due to the point-mutated Cx30 in mice...
  47. pmc Kinetic and molecular analysis of nuclear export factor CRM1 association with its cargo in vivo
    Dirk Daelemans
    Rega Institute for Medical Research, Katholieke Universiteit Leuven, Minderbroedersstraat 10, 3000 Leuven, Belgium
    Mol Cell Biol 25:728-39. 2005
    ..Using fluorescence recovery after photobleaching (FRAP), we show that CRM1 moves at rates similar to that of free green fluorescent protein in ..
  48. pmc Global nature of dynamic protein-chromatin interactions in vivo: three-dimensional genome scanning and dynamic interaction networks of chromatin proteins
    Robert D Phair
    BioInformatics Services, Rockville, MD 20854, USA
    Mol Cell Biol 24:6393-402. 2004
    ..We suggest that these properties are crucial for generating high plasticity in genome expression...
  49. pmc Analysis of binding at a single spatially localized cluster of binding sites by fluorescence recovery after photobleaching
    Brian L Sprague
    Laboratory of Receptor Biology and Gene Expression, NCI, National Institutes of Health, Bethesda, Maryland, USA
    Biophys J 91:1169-91. 2006
    ..Binding interactions within such clusters may be analyzed in live cells using models for fluorescence recovery after photobleaching (FRAP)...
  50. pmc Actin-depolymerizing factor and cofilin-1 play overlapping roles in promoting rapid F-actin depolymerization in mammalian nonmuscle cells
    Pirta Hotulainen
    Program in Cellular Biotechnology, Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland
    Mol Biol Cell 16:649-64. 2005
    ..b>Fluorescence recovery after photobleaching analysis and studies with an actin monomer-sequestering drug, latrunculin-A, demonstrated that ..
  51. ncbi Human connexin26 and connexin30 form functional heteromeric and heterotypic channels
    Sabrina W Yum
    Section of Neurology, St Christopher s Hospital For Children, Erie Ave at Front St, Philadelphia, PA 19134, USA
    Am J Physiol Cell Physiol 293:C1032-48. 2007
    ..of calcein (an anionic tracer) transfer was intermediate between their homotypic counterparts by fluorescence recovery after photobleaching. Fluorescence recovery after photobleaching also showed that Cx26 and Cx30 form functional ..
  52. ncbi Targeting of voltage-gated potassium channel isoforms to distinct cell surface microdomains
    Kristen M S O'Connell
    Department of Biomedical Sciences, Colorado State University, Ft Collins, CO 80523, USA
    J Cell Sci 118:2155-66. 2005
    ..1 expressed in HEK cells exhibits a surface distribution similar to that seen in native cells...
  53. pmc Differentiation driven changes in the dynamic organization of Basal transcription initiation
    Giuseppina Giglia-Mari
    Department of Genetics, Erasmus MC, Rotterdam, The Netherlands
    PLoS Biol 7:e1000220. 2009
    ....
  54. ncbi Using FRAP and mathematical modeling to determine the in vivo kinetics of nuclear proteins
    Gustavo Carrero
    Department of Mathematical and Statistical Sciences, University of Alberta, Alberta, Canada
    Methods 29:14-28. 2003
    b>Fluorescence recovery after photobleaching (FRAP) has become a popular technique to investigate the behavior of proteins in living cells...
  55. ncbi Fluorescence recovery after photobleaching studies of lipid rafts
    Anne K Kenworthy
    Dept of Molecular Physiology and Biophysics, Vanderbilt School of Medicine, Nashville, TN 37232, USA
    Methods Mol Biol 398:179-92. 2007
    b>Fluorescence recovery after photobleaching (FRAP) is a microscopy-based technique that can be used to ask how lipid rafts impact protein and lipid diffusion in cells...
  56. pmc Dynamics of the alpha6beta4 integrin in keratinocytes
    Cecile A W Geuijen
    Division of Cell Biology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands
    Mol Biol Cell 13:3845-58. 2002
    ..This clustering ultimately determines whether alpha6beta4 will inhibit cell migration or not...
  57. ncbi Cortical actin turnover during cytokinesis requires myosin II
    Minakshi Guha
    Department of Physiology, University of Massachussetts Medical School, Worcester, 01605, USA
    Curr Biol 15:732-6. 2005
    ..Our observations indicate that myosin II ATPase is not required for the assembly of equatorial cortex during cytokinesis but is essential for its subsequent turnover and remodeling...
  58. pmc Phase coexistence and connectivity in the apical membrane of polarized epithelial cells
    Doris Meder
    Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany
    Proc Natl Acad Sci U S A 103:329-34. 2006
    ..connectivity was assessed by measuring long-range diffusion of several membrane proteins by fluorescence recovery after photobleaching in two polarized epithelial cell lines and one fibroblast cell line...
  59. ncbi Caveolin-1 loss of function accelerates glucose transporter 4 and insulin receptor degradation in 3T3-L1 adipocytes
    Elena González-Muñoz
    Departament de Bioquimica i Biologia Molecular, Facultat de Biologia, Universitat de Barcelona, Institute for Research in Biomedicine IRB Barcelona, Serveis Científico Tècnics, Universitat de Barcelona, 08028 Barcelona, Spain
    Endocrinology 150:3493-502. 2009
    ..We propose that caveolin-1/caveolae control insulin action in adipose cells...
  60. ncbi Gephyrin oligomerization controls GlyR mobility and synaptic clustering
    Martino Calamai
    INSERM U789 Biologie Cellulaire de Synapse and Département de Biologie, Ecole Normale Superieure, 75005 Paris, France
    J Neurosci 29:7639-48. 2009
    ..Using fluorescence recovery after photobleaching, we studied the exchange kinetics of synaptic gephyrin clusters...
  61. pmc Obstructed diffusion in phase-separated supported lipid bilayers: a combined atomic force microscopy and fluorescence recovery after photobleaching approach
    Timothy V Ratto
    Biophysics Graduate Group, Division of Biological Sciences, University of California Davis, One Shields Avenue, Davis, CA 95616, USA
    Biophys J 83:3380-92. 2002
    ..b>Fluorescence recovery after photobleaching was used to measure the lateral diffusion coefficient in single supported bilayers composed of ..
  62. pmc Cdt1 associates dynamically with chromatin throughout G1 and recruits Geminin onto chromatin
    Georgia Xouri
    Laboratory of General Biology, School of Medicine, University of Patras, Patras, Greece
    EMBO J 26:1303-14. 2007
    ..We propose that dynamic Cdt1-chromatin associations and the recruitment of Geminin to chromatin provide spatio-temporal control of the licensing process...
  63. pmc Myofibrillogenesis in skeletal muscle cells in zebrafish
    Joseph W Sanger
    Department of Cell and Developmental Biology, SUNY Upstate Medical University, Syracuse, New York, USA
    Cell Motil Cytoskeleton 66:556-66. 2009
    ..b>Fluorescence Recovery After Photobleaching showed that the exchange of proteins (actin, alpha-actinin, FATZ, myotilin and telethonin) ..
  64. ncbi Dynamic in vivo interaction of DDB2 E3 ubiquitin ligase with UV-damaged DNA is independent of damage-recognition protein XPC
    Martijn S Luijsterburg
    Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, Nuclear Organisation Group, University of Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands
    J Cell Sci 120:2706-16. 2007
    ..We propose a scenario in which DDB2 prepares UV-damaged chromatin for assembly of the NER complex...
  65. pmc Intrinsic dynamic behavior of fascin in filopodia
    YVONNE S ARATYN
    Department of Cell and Molecular Biology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA
    Mol Biol Cell 18:3928-40. 2007
    ..b>Fluorescence recovery after photobleaching analysis revealed that fascin rapidly dissociates from filopodial filaments with a kinetic off-..
  66. pmc Analyzing intracellular binding and diffusion with continuous fluorescence photobleaching
    Malte Wachsmuth
    Division Biophysics of Macromolecules, German Cancer Research Center, D 69120 Heidelberg, Germany
    Biophys J 84:3353-63. 2003
    ..The cells show fluorescent nucleoli, and binding is transient. CP yields residence times for mTTF-I-EGFP of approximately 13 s...
  67. pmc Quantitative kinetic study of the actin-bundling protein L-plastin and of its impact on actin turn-over
    Ziad Al Tanoury
    Laboratory of Cytoskeleton and Cell Plasticity, Life Sciences Research Unit, University of Luxembourg, Luxembourg City, Luxembourg
    PLoS ONE 5:e9210. 2010
    ..Phosphorylation of L-plastin on residue Ser5 increases its F-actin binding activity and is required for L-plastin-mediated cell invasion...
  68. pmc Ras diffusion is sensitive to plasma membrane viscosity
    J Shawn Goodwin
    Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA
    Biophys J 89:1398-410. 2005
    ..Using confocal fluorescence recovery after photobleaching, we examined the diffusion of green fluorescent protein (GFP)-tagged HRas, NRas, and KRas in COS-..
  69. pmc Monitoring chaperone engagement of substrates in the endoplasmic reticulum of live cells
    Erik L Snapp
    Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, 18 Library Drive, Building 18, Room 101, Bethesda, MD 20892, USA
    Proc Natl Acad Sci U S A 103:6536-41. 2006
    ..Our findings provide insight into the normal organization and dynamics of an ER chaperone and characterize the capacity of the ER to maintain homeostasis during acute changes in chaperone activity and availability...
  70. pmc Modeling fluorescence recovery after photobleaching in loaded bone: potential applications in measuring fluid and solute transport in the osteocytic lacunar-canalicular system
    Xiaozhou Zhou
    Department of Mechanical Engineering, Center for Biomedical Engineering Research, University of Delaware, 126 Spencer Laboratory, Newark, DE 19716, USA
    Ann Biomed Eng 36:1961-77. 2008
    ..system is essential for osteocyte viability and function, and it can be measured using fluorescence recovery after photobleaching (FRAP)...
  71. ncbi Knee-loading modality drives molecular transport in mouse femur
    Min Su
    Department of Anatomy and Cell Biology, Indiana University Purdue University, Indianapolis, 46202, USA
    Ann Biomed Eng 34:1600-6. 2006
    ..order to evaluate load-driven molecular transport in a lacunocanalicular network, we conducted fluorescence recovery after photobleaching (FRAP) experiments using lacunae stained with uranine (376 Da)...
  72. ncbi Fibrillarin and Nop56 interact before being co-assembled in box C/D snoRNPs
    Tanguy Lechertier
    Institut Jacques Monod, UMR 7592 CNRS Universités Paris 6 et 7, 2 place Jussieu, 75251 Paris Cedex 05, France
    Exp Cell Res 315:928-42. 2009
    ..In addition, no RNA seems required to maintain fibrillarin-Nop56 interaction...
  73. ncbi Measurement of diffusion within the cell wall in living roots of Arabidopsis thaliana
    Eric M Kramer
    Biology Department, University of Massachusetts, 611 N Pleasant St, Amherst, MA 01003, USA
    J Exp Bot 58:3005-15. 2007
    ..The diffusion coefficient, D(cw), of CF in the cell wall was probed using two protocols: fluorescence recovery after photobleaching and fluorescence loss following perfusion with dye-free solution...
  74. ncbi Trapped in action: direct visualization of DNA methyltransferase activity in living cells
    Lothar Schermelleh
    Ludwig Maximilians University Munich, Department of Biology II, Planegg Martinsried, Germany
    Nat Methods 2:751-6. 2005
    ....
  75. ncbi A cytoskeletal-based perimeter fence selectively corrals a sub-population of cell surface Kv2.1 channels
    Michael M Tamkun
    Department of Biomedical Sciences, Colorado State University, Ft Collins, CO 80523, USA
    J Cell Sci 120:2413-23. 2007
    ..1 clusters are formed by sub-membrane cytoskeletal structures that limit the lateral diffusion of only the sub-population of Kv2.1 channels carrying the appropriate modifications on the Kv2.1 C-terminus...
  76. ncbi Fluorescence recovery after photobleaching reveals the biochemistry of nucleocytoplasmic exchange
    Ranieri Bizzarri
    NEST, Scuola Normale Superiore and Istituto Nanoscienze CNR, Pisa, Italy
    Anal Bioanal Chem 403:2339-51. 2012
    b>Fluorescence recovery after photobleaching (FRAP) can help unveil subtle dynamical and biochemical properties of intracellular components...
  77. ncbi A mathematical model to determine molecular kinetic rate constants under non-steady state conditions using fluorescence recovery after photobleaching (FRAP)
    Tanmay P Lele
    Vascular Biology Program, Karp Family Research Laboratories, 11 127, Children s Hospital and Harvard Medical School, 300 Longwood Avenue, Boston, MA 02115, USA
    Biophys Chem 120:32-5. 2006
    b>Fluorescence recovery after photobleaching (FRAP) analyses of binding and unbinding of molecules that interact with insoluble scaffolds, such as the cytoskeleton and nuclear matrix, in living cells commonly assume that this process is at ..
  78. ncbi Conditions for using FRAP as a quantitative technique--influence of the bleaching protocol
    Dominika O Trembecka
    Faculty of Biochemistry, Biophysics and Biotechnology, Division of Cell Biophysics, Jagiellonian University, Krakow, Poland
    Cytometry A 77:366-70. 2010
    b>Fluorescence recovery after photobleaching (FRAP) is a tool widely used in studies of dynamic behavior of fluorescently-tagged proteins in live cells...
  79. pmc The tight junction protein complex undergoes rapid and continuous molecular remodeling at steady state
    Le Shen
    Department of Pathology, The University of Chicago, Chicago, IL 60637, USA
    J Cell Biol 181:683-95. 2008
    ..tight junction protein dynamics in live confluent Madin-Darby canine kidney monolayers using fluorescence recovery after photobleaching and related methods...
  80. ncbi Fluorescence recovery after photobleaching
    Alex Carisey
    Faculty of Life Sciences, The University of Manchester, Manchester, UK
    Methods Mol Biol 769:387-402. 2011
    This chapter describes the use of microscope-based fluorescence recovery after photobleaching (FRAP)...
  81. ncbi Monitoring dynamic binding of chromatin proteins in vivo by fluorescence recovery after photobleaching
    Florian Mueller
    Groupe Imagerie et Modélisation, Institut Pasteur, CNRS, URA 2582, Paris, France
    Methods Mol Biol 833:153-76. 2012
    b>Fluorescence recovery after photobleaching (FRAP) has now become widely used to investigate nuclear protein binding to chromatin in live cells...
  82. pmc In vitro FRAP reveals the ATP-dependent nuclear mobilization of the exon junction complex protein SRm160
    Stefan Wagner
    Dept of Cell Biology, S7 214, University of Massachusetts Medical School, 55 Lake Ave, Worcester, MA 01655, USA
    J Cell Biol 164:843-50. 2004
    ..binding and mobility of enhanced green fluorescent protein (EGFP)-labeled nuclear proteins by fluorescence recovery after photobleaching in digitonin-permeabilized cells...
  83. pmc Uptake, distribution and diffusivity of reactive fluorophores in cells: implications toward target identification
    Christopher W Cunningham
    Department of Medicinal Chemistry and Pharmaceutical Chemistry, Molecular Graphics and Modeling Laboratory, and Specialized Chemistry Center, University of Kansas, Lawrence, Kansas, USA
    Mol Pharm 7:1301-10. 2010
    ..The intracellular diffusion coefficients were measured using a fluorescence recovery after photobleaching (FRAP)-based method...
  84. ncbi Mobility of model proteins in hydrogels composed of oppositely charged dextran microspheres studied by protein release and fluorescence recovery after photobleaching
    Sophie R Van Tomme
    Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences UIPS, University Utrecht, The Netherlands
    J Control Release 110:67-78. 2005
    ..These results show that these hydrogels are very suitable as injectable matrix for diffusion-controlled delivery of pharmaceutically active proteins...
  85. ncbi Fully automated attenuation measurement and motion correction in FLIP image sequences
    Martijn van de Giessen
    Division of Image Processing LKEB, Leiden University Medical Center, 2300 RC Leiden, The Netherlands
    IEEE Trans Med Imaging 31:461-73. 2012
    ..The proposed method is fully automatic, and runs in approximately one minute per sequence, making it suitable for unsupervised batch processing of large data series...
  86. pmc A generalization of theory for two-dimensional fluorescence recovery after photobleaching applicable to confocal laser scanning microscopes
    Minchul Kang
    Department of Molecular Physiology and Biophysics, Vanderbilt University School of Medicine, Nashville, Tennessee, USA
    Biophys J 97:1501-11. 2009
    b>Fluorescence recovery after photobleaching (FRAP) using confocal laser scanning microscopes (confocal FRAP) has become a valuable technique for studying the diffusion of biomolecules in cells...
  87. pmc Are assumptions about the model type necessary in reaction-diffusion modeling? A FRAP application
    Juliane Mai
    Department of Computational Hydrosystems, UFZ Helmholtz Centre for Environmental Research, Leipzig, Germany
    Biophys J 100:1178-88. 2011
    At present, fluorescence recovery after photobleaching (FRAP) data are interpreted using various types of reaction-diffusion (RD) models: the model type is usually fixed first, and corresponding model parameters are inferred subsequently...
  88. ncbi FRAP analysis of molecular diffusion inside sea-urchin spermatozoa
    Daisuke Takao
    Department of Life Science, Graduate School of Arts and Sciences, The University of Tokyo, Komaba 3 8 1, Meguro, Tokyo 153 8902, Japan
    J Exp Biol 211:3594-600. 2008
    ..Using a FRAP (fluorescence recovery after photobleaching) technique, we determined the diffusion coefficients of fluorescein-derivatives (calcein, ..
  89. pmc A closed-form analytic expression for FRAP formula for the binding diffusion model
    Minchul Kang
    Biophys J 95:L13-5. 2008
    ..b>Fluorescence recovery after photobleaching (FRAP) can be used to determine these kinetic constants in living cells...
  90. ncbi Role of three-dimensional bleach distribution in confocal and two-photon fluorescence recovery after photobleaching experiments
    Davide Mazza
    Laboratory for Advanced Microscopy, Bioimaging, and Spectroscopy MicroSCoBiO Research Center, Department of Physics, University of Genoa, Italy
    Appl Opt 46:7401-11. 2007
    The quantitative analysis of fluorescence perturbation experiments such as fluorescence recovery after photobleaching (FRAP) requires suitable analytical models to be developed...
  91. pmc Fluorescence recovery after photobleaching and photoconversion in multiple arbitrary regions of interest using a programmable array microscope
    Guy M Hagen
    Laboratory of Cellular Dynamics, Max Planck Institute for Biophysical Chemistry, Gottingen, Germany
    Microsc Res Tech 72:431-40. 2009
    ..b>Fluorescence recovery after photobleaching (FRAP) is commonly used for the determination of lateral diffusion constants of membrane ..
  92. ncbi Fluorescence recovery after photobleaching on the confocal laser-scanning microscope: generalized model without restriction on the size of the photobleached disk
    Nick Smisdom
    Hasselt University, Transnational University Limburg, Biomedical Research Institute, School of Life Sciences, Agoralaan Building C, Diepenbeek 3590, Belgium
    J Biomed Opt 16:046021. 2011
    b>Fluorescence recovery after photobleaching (FRAP) carried out on a confocal laser-scanning microscope (CLSM) performs well for photobleached disks that are large compared to the resolution of the bleaching beam...
  93. pmc Dominant-negative mechanism of leukemogenic PAX5 fusions
    N Kawamata
    Hematology Oncology, Cedars Sinai Medical Center UCLA School of Medicine, Los Angeles, CA, USA
    Oncogene 31:966-77. 2012
    ..PAX5-C20S is a tetramer, which interacts extraordinarily stably with chromatin as determined by Fluorescence Recovery After Photobleaching in living cells...
  94. pmc Evaluation of pulsed-FRAP and conventional-FRAP for determination of protein mobility in prokaryotic cells
    Jacek T Mika
    Department of Biochemistry, Groningen Biomolecular Science and Biotechnology Institute, Netherlands Proteomics Centre and Zernike Institute for Advanced Materials, University of Groningen, Groningen, The Netherlands
    PLoS ONE 6:e25664. 2011
    Macromolecule mobility is often quantified with Fluorescence Recovery After Photobleaching (FRAP)...
  95. pmc Fatty acid 2-hydroxylase mediates diffusional mobility of Raft-associated lipids, GLUT4 level, and lipogenesis in 3T3-L1 adipocytes
    Lin Guo
    Department of Internal Medicine, Center for Human Nutrition, Washington University School of Medicine, St Louis, Missouri 63110, USA
    J Biol Chem 285:25438-47. 2010
    ..of FA2H appeared to increase raft lipid mobility as it significantly accelerated the rates of fluorescence recovery after photobleaching measurements of lipid rafts labeled with Alexa 488-conjugated cholera toxin subunit B...
  96. pmc FRAP analysis of membrane-associated proteins: lateral diffusion and membrane-cytoplasmic exchange
    Nathan W Goehring
    Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
    Biophys J 99:2443-52. 2010
    Obtaining quantitative kinetic parameters from fluorescence recovery after photobleaching (FRAP) experiments generally requires a theoretical analysis of protein mobility and appropriate solutions for FRAP recovery derived for a given ..
  97. ncbi Effects of recombinant protein expression on green fluorescent protein diffusion in Escherichia coli
    Kristin M Slade
    Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599 3290, USA
    Biochemistry 48:5083-9. 2009
    b>Fluorescence recovery after photobleaching was used to measure the diffusion coefficient of green fluorescent protein (GFP, 27 kDa) in Escherichia coli in the presence or absence of four coexpressed proteins: cytoplasmic maltose binding ..
  98. ncbi Gap junctional intercellular communication capacity by gap-FRAP technique: a comparative study
    Muriel Abbaci
    Faculte de Medecine, Nancy University, Vandoeuvre les Nancy, France
    Biotechnol J 2:50-61. 2007
    ..in monolayer cultures, characterized by different patterns of phosphorylated Cx43, we used a fluorescence recovery after photobleaching (FRAP) technique, and compared two tracers, 5(6)-carboxyfluorescein diacetate (CFDA) and calcein ..
  99. ncbi Intranuclear ataxin1 inclusions contain both fast- and slow-exchanging components
    David L Stenoien
    Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030, USA
    Nat Cell Biol 4:806-10. 2002
    ..Here, we used fluorescence recovery after photobleaching (FRAP) to examine the intranuclear dynamics of polyglutamine-expanded ataxin1 and inclusion-..
  100. pmc Sorting of lipids and proteins in membrane curvature gradients
    A Tian
    Department of Chemistry, University of Pennsylvania, Philadelphia, Pennsylvania, USA
    Biophys J 96:2676-88. 2009
    ..The sorting of Cholera toxin subunit B is rationalized by statistical models. We discuss the implications of our findings for intracellular sorting mechanisms...
  101. pmc Investigating complexity of protein-protein interactions in focal adhesions
    Tanmay P Lele
    Department of Chemical Engineering, University of Florida, Gainesville, FL 32611, USA
    Biochem Biophys Res Commun 369:929-34. 2008
    ..Here, we used the fluorescence recovery after photobleaching (FRAP) technique, combined with mathematical modeling and scaling analysis to quantify ..

Research Grants64

  1. Function and Expression of Connexins in the pre-Botzinger Complex
    JONATHAN KELTY; Fiscal Year: 2007
    ..carbenoxolone, or CBX) and a control agent (glycyrrhizic acid or GZA), will be evaluated by a fluorescence recovery after photobleaching (FRAP) microscopy technique...
  2. Novel TIRF microscopy analyzing trafficking & signaling at the cell cortex
    Derek Toomre; Fiscal Year: 2009
    ..Consistent with the recovery act's goal, this funding will provide full-time summer employment for 4 undergraduate students and accelerate scientific achievement of the parent DP2 award...
  3. STRUCTURE AND FUNCTION OF PAXILLIN
    Christopher E Turner; Fiscal Year: 2012
    ..proteins in fibroblasts and utilize confocal fluorescence time-lapse microscopy, combined with Fluorescence Recovery after Photobleaching (FRAP) and Fluorescence Resonance Energy Transfer (FRET) analysis to evaluate cell morphology, ..
  4. The Role of Connexin32 in the Pathogensis of CMTX
    STEVEN SIMON SCHERER; Fiscal Year: 2013
    ..mutant to form functional GJ plaques will be investigated - by immunostaining, scrape loading, fluorescence recovery after photobleaching (FRAP), and electrophysiology...
  5. Role of Cofilin and Wnt Signaling in Neural Crest Development
    O apos NEIL WIGGAN; Fiscal Year: 2013
    ..Dynamic F-actin remodeling will be assessed by techniques such as fluorescence recovery after photobleaching. Furthermore, by use of commonly utilized biochemical procedures we will establish the means by ..
  6. Dynamics of Transcriptional Regulation
    Elena Zelin; Fiscal Year: 2011
    ..Establish the effects of p23, GCN5, and HDAC1 on transcription factor dynamics in vivo. The fluorescence recovery after photobleaching (FRAP) technique will be used in conjunction with engineered cell lines that contain hsp70 loci ..
  7. Physiology of Class III PI 3-kinase Signaling 2
    Jonathan M Backer; Fiscal Year: 2013
    ..Aim 2 examines the dynamics of hVps34 complex formation, both at the whole cell level and, using fluorescence recovery after photobleaching (FRAP), on the autophagosomal membrane...
  8. Advanced Immunocytochemistry, In Situ Hybridization and Live Cell Imaging Course
    TERRI I GRODZICKER; Fiscal Year: 2013
    ..to localize, quantify, and analyze proteins in living cells using standard fluorescence, fluorescence recovery after photobleaching, and resonance energy transfer...
  9. Nikon TiE Motorized Microscope System For 3D Image Acquisition, Deconvolution, Li
    Robert S Fuller; Fiscal Year: 2010
    ..sensitivity and resolution for 3D image acquisition, deconvolution, and live cell time lapse and fluorescence recovery after photobleaching (FRAP)...
  10. Real-time microviscosity measurement tools for the cell
    Mark A Haidekker; Fiscal Year: 2010
    ..Presently, viscosity on the microscopic scale is determined by fluorescence anisotropy, fluorescence recovery after photobleaching (FRAP), or magnetic nanoparticles...
  11. Genesis of Epithelial Cell Lumens
    Jennifer L Harder; Fiscal Year: 2013
    ..knockdown and mutagenesis, 3-dimensional cyst assay, custom cell surface trafficking assay, and fluorescence recovery after photobleaching. In Aim #2, she will evaluate an as yet unexplored mechanism of controlling polarity, that cells ..
  12. Spinning Disk Confocoal Microscope
    Judy Drazba; Fiscal Year: 2010
    ..will also provide the ability to do fluorescence resonance energy transfer (FRET) measurements, fluorescence recovery after photobleaching (FRAP) measurements and photoactivation, thereby substantially advancing their NIH-funded ..
  13. Optical studies of the cone photoreceptor synapse
    Richard H Kramer; Fiscal Year: 2013
    ..will use fluorescent markers of synaptic vesicles to measure vesicle mobility on the ribbon with Fluorescence Recovery After Photobleaching (FRAP) and Fluorescence Correlation Spectroscopy (FCS)...
  14. Molecular Mechanisms of PTH-Mediated Trafficking in Renal Tubular Cells
    Judith T Blaine; Fiscal Year: 2013
    ..in living cells will be examined using total internal reflection fluorescence microscopy and fluorescence recovery after photobleaching. These imaging techniques will be complemented by investigation of the effects of PTH on the ..
  15. The Role of Small Ankyrin 1 in the Sarcoplasmic Reticulum in Skeletal Muscle
    Andrew P Ziman; Fiscal Year: 2011
    ..flexor digitorum brevis muscle of the rat, including assays of the continuity of the stores by fluorescence recovery after photobleaching (FRAP), and Ca2+ release by activators of the ryanodine receptor and by electrical stimulation;..
  16. Analysis of Z-band assembly and maintenance in living skeletal muscle cells
    JOSEPH WILLIAM SANGER; Fiscal Year: 2013
    ..FRAP (Fluorescence Recovery After Photobleaching) experiments will measure the changing dynamics of key proteins in the z-bodies and Z-bands ..
  17. An Animal Model for Cornelia de Lange Syndrome
    DALE L DORSETT; Fiscal Year: 2012
    ..how changes in cohesion factors affect cohesin chromosome-binding dynamics in vivo using fluorescence recovery after photobleaching. It is hoped that insights from these studies will shed light on the mechanisms by which small ..
  18. SIGNAL TRANSDUCTION AT FERTILIZATION
    Laurinda A Jaffe; Fiscal Year: 2012
    ..live follicle-enclosed oocytes with fluorescent tracers, and analysis by 2-photon microscopy and fluorescence recovery after photobleaching. These studies will be complemented by investigations of the phosphorylation of connexin 43 on ..
  19. Control of Vascular Cell Motility by CaMKII
    Harold A Singer; Fiscal Year: 2013
    ..b>Fluorescence recovery after photobleaching (FRAP) analysis of GFP-tagged focal adhesion components will be used to assess CaMKII4-dependent ..
  20. Biophysics Cell Membrane Probe
    William E Brownell; Fiscal Year: 2010
    ..The design is modular so that a variety of user specific experiments such as calcium imaging, fluorescence recovery after photobleaching or fluorescence resonance energy transfer may be easily accommodated...
  21. MECHANICAL LOADING AND BONE
    Hiroki Yokota; Fiscal Year: 2013
    ..strains using electronic speckle pattern interferometry as well as molecular transport using fluorescence recovery after photobleaching;(3) conducting bone histomorphometry to evaluate ex vivo data;and (4) examining load-driven ..
  22. ZEISS LSM710 SCANNING CONFOCAL MICROSCOPE
    Hazel L Sive; Fiscal Year: 2010
    ..The LSM710 can perform FRAP (Fluorescence Recovery After Photobleaching) to analyze molecular dynamics;calcium imaging is simple and targeted uncaging of fluorescent ..
  23. Reticulon Function in ER Morphology
    Gia Voeltz; Fiscal Year: 2013
    ..For these studies, we will be using fluorescence microscopy, immunoprecipitations, fluorescence recovery after photobleaching, and electron microscopy with immunogold labeling...
  24. In vivo and crude extract analysis of polyQ aggregation intermediates
    Robert Fairman; Fiscal Year: 2012
    ..This technique will be used, combined with FRAP (fluorescence recovery after photobleaching, carried out using a confocal microscope), to probe the aggregation states of polyQ-containing ..
  25. Solute Transport in the Bone Lacunar-Canalicular System
    Liyun Wang; Fiscal Year: 2012
    ..To this end, we recently developed a new imaging method based on Fluorescence Recovery After Photobleaching (FRAP) that allows measurement of solute movement in the bone LCS in situ and in real-time (Wang ..
  26. Regulation of ENaC by phosphoinositides, MARCKS, and the cytoskeleton
    ABDEL AYUBE ALLI; Fiscal Year: 2012
    ..We will perform Fluorescence Recovery after Photobleaching (FRAP) to determine the role of the actin cytoskeleton in the formation of a membrane signaling ..
  27. Biophysical Basis of Organization and Dynamics of Prestin Membrane Complexes
    Robert M Raphael; Fiscal Year: 2013
    ..including fluorescence resonance energy transfer (FRET), fluorescence lifetime imaging (FLIM), fluorescence recovery after photobleaching (FRAP) and single molecule imaging...
  28. MOLECULAR MECHANISM OF PHOTORECEPTOR G PROTEIN SIGNALING
    Nikolai O Artemyev; Fiscal Year: 2013
    ..The mutant Gat1 models will be examined with EGFP imaging, immunofluorescence, and Fluorescence Recovery After Photobleaching (FRAP) analysis of lateral and longitudinal diffusion...
  29. Regulation of activity-dependent ProSAp2 synaptic dynamics
    MAGALI HOLT ROWAN; Fiscal Year: 2011
    ..Live-cell time-lapse confocal microscopy and Fluorescence Recovery After Photobleaching (FRAP) will be used here to assess the dynamic exchange properties of ProSAP2 in response to ..
  30. Columbia AFM System Equipment Grant
    Clark T Hung; Fiscal Year: 2010
    ..module provides photoactivation or bleaching and simultaneous image collection, enabling fluorescence recovery after photobleaching (FRAP) and molecular uncaging applications...
  31. P2X channel mobility in the plasma membrane of neurons
    Esther Richler; Fiscal Year: 2011
    ..The specific goals of this proposal are 1) to use fluorescence recovery after photobleaching (FRAP) and single particle tracking (SPT) of Quantum dot labelled P2X2 receptees to probe the ..
  32. Automated Pre-screening System for in meso Membrane protein crystallization
    VADIM GENNADYEVICH CHEREZOV; Fiscal Year: 2012
    ..We propose to develop an automated system for carrying out Fluorescence Recovery after Photobleaching (FRAP) studies of membrane proteins in meso...
  33. Automated Pre-screening System for in meso Membrane protein crystallization
    VADIM GENNADYEVICH CHEREZOV; Fiscal Year: 2011
    ..We propose to develop an automated system for carrying out Fluorescence Recovery after Photobleaching (FRAP) studies of membrane proteins in meso...
  34. Tumor cell and microenvironment changes causing antiangiogenic therapy resistance
    Manish Aghi; Fiscal Year: 2013
    ..novel in vivo models of anti-angiogenic therapy resistance and an innovative application of fluorescence recovery after photobleaching (FRAP) to correlate integrin mobility and turnover in focal adhesions with drug resistance...
  35. INTRACELLULAR TRANSPORT OF AMPHIPATHS
    Bruce Luxon; Fiscal Year: 2000
    ..and a fluorescent bile acid analog dansyl taurocholate, will be measured by two dimensional fluorescence recovery after photobleaching (FRAP)...
  36. Multiphoton Laser Scanning Microscope: Zeiss LSM710 NLO
    Olga Kovbasnjuk; Fiscal Year: 2010
    ..including advanced imaging applications which include Forster Resonance Energy Transfer (FRET), Fluorescence Recovery After Photobleaching (FRAP), deconvolution, Second Harmonic Generation (SHG), quantitative imaging protocols for pH-..
  37. GPCR signaling complexes in living cells
    NEVIN ALAN LAMBERT; Fiscal Year: 2010
    ..cells, and the lateral mobility of potentially interacting proteins is measured by monitoring fluorescence recovery after photobleaching. We will use this method together with standard electrophysiological techniques to test specific ..
  38. Functional and Pharmacological Implications of mGluR Heteromerization
    Paul J Kammermeier; Fiscal Year: 2013
    ..together to form dimeric, tetrameric, or higher order multimeric receptors using multi-photon Fluorescence Recovery After Photobleaching (FRAP) and Fluorescence Correlation Spectroscopy (FCS), and 4) To verify the physiological ..
  39. Hair-Bundle Lipids
    Peter Gillespie; Fiscal Year: 2009
    ..In Aim 2, using fluorescence recovery after photobleaching, we will determine dynamics of the bulk hair-bundle membrane as well as the dynamics of ..
  40. Acquisition of a Confocal Microscope for R.M Bock Laboratories
    KEVIN WILLIAM ELICEIRI; Fiscal Year: 2010
    ..fluorescence studies in 2D and 3D to more advanced applications such as photoactivation, fluorescence recovery after photobleaching (FRAP), and fluorescence resonance energy transfer (FRET)...
  41. Novel Epigenetic Marks in Mitosis
    KENNETH S contact ZARET; Fiscal Year: 2010
    ..Our recent mechanistic studies using fluorescence recovery after photobleaching (FRAP) show that FoxA and PARP binding to chromatin is far more stable than that of most other ..
  42. SPERM MEMBRANE CHANGES WITH MATURATION AND CAPACITATION
    David Wolf; Fiscal Year: 1990
    ..to understand how these restrictions change with maturation and capacitation, we propose to use fluorescence recovery after photobleaching, intensified video fluorescence microscopy, and rapid freeze deep etch electron microscopy to ..
  43. Confocal Microscopy Imaging System
    Murad Ookhtens; Fiscal Year: 2007
    ..such as live cell and time-lapse imaging, imaging specimens labeled with 3 or 4 fluorophores, fluorescence recovery after photobleaching, or fluorescence resonance energy transfer, which are sorely needed by our user base...
  44. BIOMEDICAL RESEARCH SUPPORT
    SAMUEL SHACKS; Fiscal Year: 1991
    ..the laser scanning confocal microscope system will complement existing instrumentation for fluorescence recovery after photobleaching (FRAP) and multiparameter digitized video microscopy (MDVM) and will substantially extend our ..
  45. LASER SCANNING CONFOCAL MICROSCOPE
    Peter Gillespie; Fiscal Year: 2009
    ..Olympus, Zeiss 510 Meta, and Nikon C1si, we selected Olympus because of use of its ability to fluorescence recovery after photobleaching (FRAP) with excellent time resolution;quality of images obtained during demonstrations;its ..
  46. MYOFIBRILLOGENESIS IN LIVING CARDIAC MUSCLE CELLS
    Joseph Sanger; Fiscal Year: 2006
    ..Images of live cells will be analyzed by optical techniques, such as Fluorescence Recovery after Photobleaching (FRAP), confocal and deconvolution microscopy...
  47. MITOSIS IN NORMAL AND NEOPLASTIC CELLS
    BILL BRINKLEY; Fiscal Year: 2006
    ..Additional experiments and novel methodology will be introduced such as Fluorescence Recovery After Photobleaching (FRAP) to track the movement and dynamics of bioluminescent proteins in live cells and new FIAsH-..
  48. ZEISS LSM 510 CONFOCAL MICROSCOPE SYSTEM
    Richard Kurten; Fiscal Year: 2005
    ..FRET, multiple color live cell tracking experiments, organelle-specific photobleaching (FRAP, fluorescence recovery after photobleaching and FLIP, fluorescence loss in photobleaching) as well as excellent standard confocal microscopy ..
  49. Olympus FV1000 Confocal Fluorescence Microscope
    James Goldenring; Fiscal Year: 2007
    ..unique capabilities for live cell investigations including enhanced experimental approaches to Fluorescence Recovery after Photobleaching (FRAP), imaging in the ultraviolet range, and the ability to utilize photoactivatable probes for ..
  50. RED CELL AQUAPORIN-1 WATER TRANSPORT PROTEIN
    Peter Agre; Fiscal Year: 2002
    ..distributions of AQP1 in red cell membranes will be undertaken with anti-Co by measurement of fluorescence recovery after photobleaching. Similar studies will be performed on enzymatically modified red cells and red cells from ..
  51. Asymmetric division as a morphological checkpoint for B. subtilis sporulation
    TINYA FLEMING; Fiscal Year: 2009
    ..Protein-protein interactions at the septum may also regulate aF and aE activity. The use of Fluorescence Recovery After Photobleaching (FRAP) will identify changes in protein mobility, which varies as a result of changes in protein ..
  52. Trafficking of NaPilla in brush border microvilli
    Judith Blaine; Fiscal Year: 2006
    ..The proposed studies will use TIR-FM microscopy coupled with fluorescence recovery after photobleaching (FRAP) and image correlation spectroscopy (ICS) techniques to examine how alterations in ..
  53. Ligand Binding Kinetics and Cell Adhesion by Fc Receptor
    Cheng Zhu; Fiscal Year: 2005
    ..Measure the kinetics of Fc-gamma-R-IgG interactions by a new contact area FRAP (fluorescence recovery after photobleaching) method...
  54. Membrane Polarization and Endothelial Cell Motility
    Paul Fox; Fiscal Year: 2007
    ..that basic fibroblast growth factor increases EC plasma membrane microviscosity as measured by fluorescence recovery after photobleaching (FRAP)...
  55. R21: DNA Electrophoresis on nanostructured surfaces
    Dilip Gersappe; Fiscal Year: 2006
    ..b>Fluorescence recovery after photobleaching (FRAP) coupled with Linear Dichroism detection (FDLD) will be used to measure surface relaxation ..
  56. Leica AOBS SP2 confocal for the MCB/DBS Imaging Facility
    Jodi Nunnari; Fiscal Year: 2004
    ..First is the inability to perform key experiments, including 1) fluorescence recovery after photobleaching (FRAP), 2) photo-activation of fluorophores in defined regions of interest, and 3) time-resolved ..
  57. BIOMEDICAL RESEARCH SUPPORT
    John Reid; Fiscal Year: 1991
    ..ACAS 570 features vital to this research include capability to measure fluorescence recovery after photobleaching, adherent live cell sorting, laser cross-linking, quantitative fluorescence photometry, laser ..
  58. MEMBRANE LIPID ORDER AND LYMPHOCYTE FUNCTION
    Robert Schlegel; Fiscal Year: 1992
    ..b>Fluorescence recovery after photobleaching will be employed to assess whether increases agglutinability of thymocytes with loosely-packed ..
  59. UPDATE IMAGING CORE FACILITY
    Katherine Luby Phelps; Fiscal Year: 1992
    ..Its four workstations for digital microscopy, laser confocal microscopy, fluorescence recovery after photobleaching (FRAP) and off-line image processing are extensively used by several laboratories...
  60. Spinning Microlens Confocal Microscope
    William Mohler; Fiscal Year: 2004
    ..dye-tunable photobleaching laser system is included for high-time/space-resolution studies of fluorescence recovery after photobleaching (FRAP), as well as dequenching measurements of fluorescence resonance energy transfer (FRET)...
  61. PURCHASE OF AN INTERACTIVE LASER CYTOMETER
    Randolph Noelle; Fiscal Year: 1993
    ..within cells by confocal imaging; 5) determine the rates and extent of antigen translocation by fluorescence recovery after photobleaching and 6) examine the biological and biochemical responses of individual cells through the use of ..
  62. OPTICAL ANALYSIS OF SYNAPTIC VESICLE RECYCLING
    WILLIAM BETZ; Fiscal Year: 2009
    ..Both Fluorescence Recovery After Photobleaching (FRAP) and Image Correlation Spectroscopy (ICS) will be employed to quantify vesicle ..
  63. Characterization of mammalian stress granules
    Paul J Anderson; Fiscal Year: 2010
    ..These aims will be accomplished by using fluorescence recovery after photobleaching analysis to quantify movement of mRNA and proteins in and out of SGs and PBs...
  64. STRUCTURAL, KINETIC, AND CELLULAR MAPPING OF THE NAT+ CH
    KIMON ANGELIDES; Fiscal Year: 1990
    ..To examine the mobility of Na+ channels during neuronal differentiation, fluorescence recovery after photobleaching will be used in an in vitro myelination system of purified spinal cord neurons and Schwann cells ..