Award Abstract #0100828
SGER - Atomic Force Microscopy System with Single Molecule Fluorescence Capabilites

NSF Org:	DBI Division of Biological Infrastructure
Initial Amendment Date:	December 5, 2000
Latest Amendment Date:	October 9, 2002
Award Number:	0100828
Award Instrument:	Standard Grant
Program Manager:	Gerald Selzer DBI Division of Biological Infrastructure BIO Directorate for Biological Sciences
Start Date:	December 15, 2000
Expires:	May 31, 2003 (Estimated)
Awarded Amount to Date:	$99756
Investigator(s):	Yuri Lyubchenko ylyubchenko@unmc.edu (Principal Investigator)
Sponsor:	Arizona State University ORSPA TEMPE, AZ 85287 480/965-5479
NSF Program(s):	INSTRUMENTAT & INSTRUMENT DEVP
Field Application(s):	0510301 Structure & Function
Program Reference Code(s):	BIOT, 9237, 9184
Program Element Code(s):	1108

ABSTRACT



Single molecule studies can provide unique information on the structure and dynamics of

biological molecules and cellular complexes that is impossible to obtain using traditional

structural techniques. These studies, to be performed at conditions close to physiological

environments, open the way for looking directly at the function of individual biological

molecules and their complexes. Scanning probe microscopy and atomic force microscope

(AFM) in particular has proven to be a very useful technique permitting static and dynamic

studies of molecules at nanometer range resolution. However, the requirement for the sample to

be bound to the surface restricts considerably the dynamical studies. A breakthrough has

recently been made in the real-time observation of dynamics of molecular complexes at the

single-molecule level using single-photon sensitive fluorescence microscopes. However, the

resolution of this technique is limited by the wavelength of the light and does not reveal

structural details of molecules. The combination of AFM and a single molecule fluorescence

(SMF) microscope into one instrument would enable us to identify structural details of

biological systems at the molecular level by AFM and to use the SMF microscope to observe

directly the dynamics of an experimental system at a the single molecule level. Importantly, the

dynamics can be performed on molecular systems structurally characterized by AFM and

localized on a comparatively large surface area. Therefore, the major objective of this proposal

is to build AFM/SMF system based on the currently available Bioscope AFM/fluorescence

microscope instrument. The sensitivity of the fluorescent system will be tested using specially

designed samples. In addition, potential pitfalls of integrated AFM will be identified and

analyzed for solutions.

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