Award Abstract #0721913
MRI: Development of Simultaneous Single Molecule Fluorescence and Atomic Force Microscopy

NSF Org:	DBI Division of Biological Infrastructure
Initial Amendment Date:	August 13, 2007
Latest Amendment Date:	August 13, 2007
Award Number:	0721913
Award Instrument:	Standard Grant
Program Manager:	Steven E. Ellis DBI Division of Biological Infrastructure BIO Directorate for Biological Sciences
Start Date:	September 1, 2007
Expires:	August 31, 2010 (Estimated)
Awarded Amount to Date:	$709945
Investigator(s):	Yale Goldman goldmany@mail.med.upenn.edu (Principal Investigator) Dennis Discher (Co-Principal Investigator)
Sponsor:	University of Pennsylvania Research Services Philadelphia, PA 19104 215/898-7293
NSF Program(s):	MAJOR RESEARCH INSTRUMENTATION
Field Application(s):
Program Reference Code(s):	BIOT, 9184, 7465, 1189
Program Element Code(s):	1189

ABSTRACT

This award is for the development of a new instrument capable of single-molecule fluorescence measurements of angular (polarization), translational, and conformational motions collected during Atomic Force Microscopy (AFM) imaging and manipulation of individual biomolecules and their complexes. AFM is the only technique that can image wet, native samples with nanometer resolution, and it has also emerged as a key technology for manipulation-type studies of single molecules and complexes, including protein and RNA folding-unfolding under tension, and studies of enzyme activity under compressive confinement. Fluorescence microscopy has also progressed over the last decade to single molecule localization considerably beyond the classical diffraction limit, and structural dynamics by intramolecular distance, spatial orientation and mobility. The goal of this award is to design and develop a new instrument that will report the internal structural changes of single macromolecules and surface layers by single molecule fluorescence microscopy, simultaneous with application and detection of relevant mechanical forces and distances by AFM. The long term objective for this new microscope is to increase fundamental understanding of assembly, folding and function of macromolecules and surface layers.

Beyond research discoveries made possible with the new microscope, the broader impacts of the award include training, outreach and communication. Students and post-doctoral fellows will be integrated into the mechanical, optical, electronic and software development. The cross-disciplinary experience of conceiving a new technology, bringing it to practical fruition, and applying it directly to a current scientific problem cannot be obtained in any other way. The new microscope will be in the Penn Nano-Bio Interface Center, which provides community outreach that educates under-represented groups and high school students and teachers.

PUBLICATIONS PRODUCED AS A RESULT OF THIS RESEARCH

Brown AE, Hategan A, Safer D, Goldman YE, Discher DE.. "Cross-correlated TIRF/AFM reveals asymmetric distribution of force-generating heads along self-assembled, "synthetic" myosin filaments.," Biophysical Journal, v.96, 2009, p. 1952.

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