The researchers characterize their new technique as a neat solution to the "needle in a haystack" problem of nanoscale microscopy, but it's more like the difference between finding the coffee table in a darkened room either by walking around until you fall over it, or using a flashlight. In a new paper,* a group from JILA—a joint venture of the National Institute of Standards and Technology (NIST) and the University of Colorado—finds tiny assemblies of biomolecules for subsequent detailed imaging by combining precision laser optics with atomic force microscopy.
The atomic force microscope (AFM) has become one of the standard tools of nanotechnology. The concept is deceptively simple. A needle—not unlike an old-fashioned phonograph stylus, but much smaller with a tip at most only a couple of atoms wide—moves across the surface of the specimen. A laser measures tiny deflections of the tip as it is pushed or pulled by atomic scale forces, such as electrostatic forces or chemical attraction. Scanning the tip back and forth across the sample yields a three-dimensional image of the surface. The resolution can be astonishing—in some cases showing individual atoms, a resolution a thousand times smaller than the best optical microscopes can achieve.
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 laser—and detection technique—is 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." ###
* A.B. Churnside, G.M. King and T.T. Perkins. Label-free optical imaging of membrane patches for atomic force microscopy. Optics Express. Vol. 18, No. 23. Nov. 8, 2010.
** The team used "purple membrane," which is cell membrane from certain single-cell organisms and contains bacteriorhodopsin, a protein that captures light energy. Bacteriorhodopsin is embedded in purple membrane and is a common protein for research in the biosciences.
Contact: Michael Baum email@example.com 301-975-2763 National Institute of Standards and Technology (NIST)