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AFM positioning: Shining light on a needle in a haystack
Date:11/10/2010

ually are "soft" things like proteins or membranes that can be damaged by an uncontrolled collision with the tip. One solution has been to "label" the target molecule with a small fluorescent compound or quantum dot, so that it lights up and is easy to find, but that means chemically altering the subject, which may not be desirable.

Instead, the JILA team opted to use a flashlight. Building upon an earlier innovation for stabilizing the position of an AFM tip, the group uses a tightly focused, low-power laser beam to optically scan the area, identifying target locations by minute changes in the scattered light. This laser is scanned across the sample to form an image, analogous to forming an AFM image.

The same laserand detection techniqueis used to locate the AFM tip. Hence, the laser serves as a common frame of reference and it's relatively straightforward to align the optical and the AFM image. In experiments with patches of cell membrane from single-cell organisms,** the group has demonstrated that they can locate these protein complexes and align the AFM tip with a precision of about 40 nanometers. Relying solely on scattered light, their technique requires no prior chemical labeling or modification of the target molecules.

"You solve a couple of problems," says NIST physicist Thomas Perkins. "You solve the problem of finding the object you want to study, which is sort of a needle in a haystack problem. You solve the problem of not contaminating your tip. And, you solve the problem of not crashing your tip into what you were looking for. This prevents damaging your tip and, for soft biological targets, not damaging your sample." And, he says, it's much more efficient. "From a practical perspective, instead of my grad student starting to do real science at 4 p.m., she can start doing science at 10 a.m."


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Contact: Michael Baum
michael.baum@nist.gov
301-975-2763
National Institute of Standards and Technology (NIST)
Source:Eurekalert  

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