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Biplane FPALM super resolution microscope

30 March 2009: Dr. Brian T. Bennett and Dr. Joerg Bewersdorf (Yale Shool of Medicine) announce completion of a BiPlane FPALM super-resolution microscope in the laboratory of Dr. Erik Jorgensen at the University of Utah
Biplane Instrumentation and Design
  

Brian Bennett
Research Assistant Professor
Biology Department
University of Utah
Bennett@biology.utah.edu

See the scope in action

Joerg Bewersdorf
Assistant Professor
Department of Cell Biology
Yale School of Medicine
Joerg.Bewersdorf@yale.edu

University of Utah, Department of Biology - Dr. Brian T. Bennett (Assistant Research Professor in the laboratory of Dr. Erik Jorgensen), in collaboration with Dr. Joerg Bewersdorf (Assistant Professor, Department of Cell Biology, Yale University), completed the assembly of a super-resolution FPALM, Biplane microscope.  This instrument is a counterpart to the recently completed system at The Jackson Laboratory. This version improves the user interface and operation through a unique adaptation of the Agilent/Till Photonics Imic microscope. Biplane microscopy provides three-dimensional sub-100nm resolution (~30X30X75nm) of thick samples without scanning. The method employs the dual plane detection system of Biplane microscopes to achieve super-resolution in the z-axis.  Biplane methodology is combined with fluorescent photoactivatable localization microscopy (FPALM) to enable 3D sub-diffraction resolution without scanning.


Bennett, Bewersdorf, Knight and Hess publications:
Bennett, B.T., Bewersdorf, J., and Knight, K.L. (2009) Immunofluorescence Imaging of DNA Damage Response Proteins: Optimizing Protocols for Super-resolution Microscopy. Journal Methods (In Press).

Juette, M.F., Gould, T.J., Lessard, M.D., Mlodzianoski, M.J., Nagpure, B.S., Bennett, B .T., Samuel T., Hess, S.T., and Bewersdorf, J. (2008) Three-Dimensional sub-100 nm Resolution Fluorescence Microscopy of Thick Samples. Nature Methods 5, 527-529

Bewersdorf, J., Bennett, B.T., & Knight, K.L. (2006) H2AX and gamma-H2AX Chromatin Structures Revealed by 4Pi Microscopy and Their Response to DNA Damage. Proc. Natl. Acad. Sci., U.S.A. 103, 18137-18142.

Bennett, B.T. and Knight, K.L. (2005) Cellular Localization of Human Rad51C and Regulation of Ubiquitin-mediated Proteolysis of Rad51. J. Cell. Biochem. 96, 1095-1109

Forget, A.L., Bennett, B.T. and Knight, K.L. (2004) Xrcc3 is Recruited to DNA Double Strand Breaks Early and Independent of Rad51. J. Cell. Biochem. 93, 429-436

Forget, A.L.*, Loftus, M.*, McGrew, D.A., Bennett, B.T. & Knight, K.L. (2007) The Human Rad51 ATP Site Mutant K133A is Functional for DNA Double-strand Repair in Human Cells. Biochemistry 46, 3566-3575.

T. J. Gould., V.V. Verkhusha., and S.T. Hess. (2009) Imaging Biological Structures with Fluorescence Photoactivation Localization Microscopy, Nature Protocols 4: 291-308

S.T. Hess. (2009) Red Lights, Camera, Photoactivation! Nature Methods 6: 124-125

T.J. Gould and S. T. Hess. (2008) Nanoscale Imaging of Intracellular Fluorescent Proteins: Breaking the Diffraction Barrier. in Biophysical Tools for Biologists, Volume 2: Methods in Vivo, H. W. Detrich editor, Methods in Cell Biology 89: 329-358

T. J. Gould., M.S. Gunewardene., M.V. Gudheti., V.V. Verkhusha., S.R. Yin., J.A. Gosse., and S.T. Hess. (2008) Imaging Molecular Positions and Anisotropies. Nature Methods 5: 1027-30

S. T. Hess., T. J. Gould., M. V. Gudheti., S.A. Maas., K.D. Mills., and J. Zimmerberg. (2007) Dynamic Clustered Distribution of Hemagglutinin Resolved at 40 nm in Living Cell Membranes Discriminates Between Raft Theories. PNAS 104: 17370-5

S. T. Hess., T.P.K. Girirajan., and M.D. Mason. (2006) Ultra-High Resolution Imaging by Fluorescence Photoactivation Localization Microscopy. Biophysical Journal 91: 4258-4272


Collaborators

Sam Hess
Assistant Professor of Physics
University of Maine
Sam Hess' group
U Maine biophysics group

UNIVERSITY OF UTAH

JORGENSEN HOMEPAGE

Kendall Knight
Professor
Department of Biochemistry and Molecular Pharmacology
UMass Medical School