Novel TIRF microscopy analyzing trafficking & signaling at the cell cortex

Summary

Principal Investigator: Derek Toomre
Abstract: In 3-month paid research experiences, over two summers, 4 talented undergraduate engineering and computer science students will be recruited to contribute in a substantial way to the progress of sub-aims of my parent New Innovator (DP2) Award; the DP2's main specific aims are to develop novel microscopes (Aim 1) and analytical tools (Aim 2) to study membrane trafficking at the cell cortex (Aim 3). Specifically, students will work on one of two projects: i) Design and construct and improve electronic circuits to control the galvometric XY mirror on our multi-angle FRAP/TIRFM (Aim 1B) and/or new auto-sensing calibration devices (Aim 2) or ii) Develop models or software to visualize and track vesicles in 3D by multi-angle TIRFM (Aim 2 ). Their advances will directly benefits goals of the DP2 and the new electronics and software's performance will be benchmarked and iteratively improved. Training and oversight will come from the PI and the DP2-supported senior scientist, Dr. Polejaev. Students will benefit from the infrastructure of the Yale 'CINEMA'lab imaging center, which is under the PI's directorship. Two outstanding US engineering students have already been identified, Noah Pestana and Isaac Anderson, both of whom have expressed a high interest in participating this summer on projects (i) and (ii), respectively. In the second summer, a particular emphasis will be made to recruit minority undergraduate students through the Yale 'STARS'program for minorities. All proposed activity are within the scope of the parent DP2 and capitalizes on early successes and will accelerate the tempo of two of the approved specific aims. 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.
Funding Period: 2009-08-03 - 2010-08-31
more information: NIH RePORT

Top Publications

  1. pmc Both daughter cells traffic and exocytose membrane at the cleavage furrow during mammalian cytokinesis
    John W Goss
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
    J Cell Biol 181:1047-54. 2008
  2. pmc Live-cell imaging of exocyst links its spatiotemporal dynamics to various stages of vesicle fusion
    Felix Rivera-Molina
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
    J Cell Biol 201:673-80. 2013
  3. pmc A network of interactions enables CCM3 and STK24 to coordinate UNC13D-driven vesicle exocytosis in neutrophils
    Yong Zhang
    Department of Pharmacology, Yale School of Medicine, New Haven, CT 06520, USA Department of Vascular Biology and Therapeutic Program, Yale School of Medicine, New Haven, CT 06520, USA
    Dev Cell 27:215-26. 2013
  4. pmc Deciphering subcellular processes in live imaging datasets via dynamic probabilistic networks
    Kresimir Letinic
    Department of Neurobiology, Yale University School of Medicine, New Haven, CT 06510, USA
    Bioinformatics 26:2029-36. 2010
  5. ncbi 3-D reconstruction of microtubules from multi-angle total internal reflection fluorescence microscopy using Bayesian framework
    Qian Yang
    Department of Electrical Engineering, Yale University, New Haven, CT 06511, USA
    IEEE Trans Image Process 20:2248-59. 2011
  6. pmc Dual-mode of insulin action controls GLUT4 vesicle exocytosis
    Yingke Xu
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
    J Cell Biol 193:643-53. 2011
  7. pmc The Legionella pneumophila effector DrrA is sufficient to stimulate SNARE-dependent membrane fusion
    Kohei Arasaki
    Section of Microbial Pathogenesis, Yale University School of Medicine, New Haven, CT 06526, USA
    Cell Host Microbe 11:46-57. 2012
  8. pmc Caveolae, fenestrae and transendothelial channels retain PV1 on the surface of endothelial cells
    Eugene Tkachenko
    Department of Medicine, University of California, San Diego, California, United States of America
    PLoS ONE 7:e32655. 2012
  9. pmc A phosphoinositide switch controls the maturation and signaling properties of APPL endosomes
    Roberto Zoncu
    Department of Cell Biology, Kavli Institute For Neuroscience and Program in Cellular Neuroscience, Neurodegeneration and Repair, Yale University School of Medicine, New Haven, CT 06510, USA
    Cell 136:1110-21. 2009
  10. ncbi Using light to see and control membrane traffic
    Yingke Xu
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520 8002, USA
    Curr Opin Chem Biol 15:822-30. 2011

Scientific Experts

  • RADU STAN
  • Derek Toomre
  • Qian Yang
  • Yingke Xu
  • Felix Rivera-Molina
  • Yong Zhang
  • Eugene Tkachenko
  • Kohei Arasaki
  • Kresimir Letinic
  • Roberto Zoncu
  • John W Goss
  • Dianqing Wu
  • Titus J Boggon
  • Kun Gao
  • Weijun Pan
  • Xiaofeng Niu
  • Wang Min
  • Jiasheng Zhang
  • Wenwen Tang
  • Haifeng Zhang
  • Sophie J Deharvengt
  • Paul F Pilch
  • Yan Xu
  • Olga Sideleva
  • Marcus R Luciano
  • Caitlin L McGarry
  • Craig R Roy
  • John Chidlow
  • Dan Tse
  • William C Sessa
  • Jonathan S Bogan
  • Alexander Karpikov
  • Thomas J Melia
  • Chenfei Yu
  • Charisse M Orme
  • Bradley R Rubin
  • Andrew Barthel
  • Rafael Sebastian
  • Pietro De Camilli
  • Michelle Pirruccello
  • Daniel M Balkin
  • Rushika M Perera

Detail Information

Publications10

  1. pmc Both daughter cells traffic and exocytose membrane at the cleavage furrow during mammalian cytokinesis
    John W Goss
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
    J Cell Biol 181:1047-54. 2008
    ..In contrast, quantitative analysis of midbody abscission showed inheritance of the midbody remnant by one daughter cell, indicating that cytokinesis is composed of both symmetrical and asymmetrical stages...
  2. pmc Live-cell imaging of exocyst links its spatiotemporal dynamics to various stages of vesicle fusion
    Felix Rivera-Molina
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA
    J Cell Biol 201:673-80. 2013
    ..Additionally, Sec8 dynamically repositioned to the site of membrane expansion, suggesting that it may respond to local cues during early cell polarization...
  3. pmc A network of interactions enables CCM3 and STK24 to coordinate UNC13D-driven vesicle exocytosis in neutrophils
    Yong Zhang
    Department of Pharmacology, Yale School of Medicine, New Haven, CT 06520, USA Department of Vascular Biology and Therapeutic Program, Yale School of Medicine, New Haven, CT 06520, USA
    Dev Cell 27:215-26. 2013
    ..Together, these findings reveal a function of the STK24 and CCM3 complex in the regulation of ligand-stimulated exocytosis. ..
  4. pmc Deciphering subcellular processes in live imaging datasets via dynamic probabilistic networks
    Kresimir Letinic
    Department of Neurobiology, Yale University School of Medicine, New Haven, CT 06510, USA
    Bioinformatics 26:2029-36. 2010
    ..Live cell imaging reveals changes in the cellular states, but current simple approaches extract only minimal information of a static snapshot...
  5. ncbi 3-D reconstruction of microtubules from multi-angle total internal reflection fluorescence microscopy using Bayesian framework
    Qian Yang
    Department of Electrical Engineering, Yale University, New Haven, CT 06511, USA
    IEEE Trans Image Process 20:2248-59. 2011
    ..The presented method has the potential to provide a reliable tool for 3-D reconstruction and tracking of microtubules...
  6. pmc Dual-mode of insulin action controls GLUT4 vesicle exocytosis
    Yingke Xu
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520, USA
    J Cell Biol 193:643-53. 2011
    ..Our results support a model whereby insulin promotes exocytic flux primarily by releasing an intracellular brake, but also by accelerating plasma membrane fusion and switching vesicle traffic between two distinct circuits...
  7. pmc The Legionella pneumophila effector DrrA is sufficient to stimulate SNARE-dependent membrane fusion
    Kohei Arasaki
    Section of Microbial Pathogenesis, Yale University School of Medicine, New Haven, CT 06526, USA
    Cell Host Microbe 11:46-57. 2012
    ....
  8. pmc Caveolae, fenestrae and transendothelial channels retain PV1 on the surface of endothelial cells
    Eugene Tkachenko
    Department of Medicine, University of California, San Diego, California, United States of America
    PLoS ONE 7:e32655. 2012
    ....
  9. pmc A phosphoinositide switch controls the maturation and signaling properties of APPL endosomes
    Roberto Zoncu
    Department of Cell Biology, Kavli Institute For Neuroscience and Program in Cellular Neuroscience, Neurodegeneration and Repair, Yale University School of Medicine, New Haven, CT 06510, USA
    Cell 136:1110-21. 2009
    ..These findings reveal a surprising plasticity of the early endocytic pathway. Importantly, PI3P functions as a switch to dynamically regulate maturation and signaling of APPL endosomes...
  10. ncbi Using light to see and control membrane traffic
    Yingke Xu
    Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06520 8002, USA
    Curr Opin Chem Biol 15:822-30. 2011
    ..Herein we highlight the utility and promise of these strategies and discuss their current limitations...