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Semiconductor Nanocrystals: The Next Thing In Fluorescent Probes

Date:
September 28, 1998
Source:
Lawrence Berkeley National Laboratory
Summary:
Some of the more shadowy secrets of biology may soon be illuminated through the use of a new type of fluorescent probe developed by scientists with the U.S. Department of Energy's Lawrence Berkeley National Laboratory and the University of California at Berkeley.

BERKELEY, CA -- Some of the more shadowy secrets of biology may soon be illuminated through the use of a new type of fluorescent probe developed by scientists with the U.S. Department of Energy's Lawrence Berkeley National Laboratory and the University of California at Berkeley.

A joint LBNL-UCB research team led by Paul Alivisatos and Shimon Weiss has announced the development of nanometer-sized crystals of semiconductors, such as cadmium selenide and cadmium sulfide, that can be used as fluorescent probes for the study of biological materials. These semiconductor nanocrystals offer a distinct advantage over conventional dye-molecules in that they emit multiple colors of light, which means they can be used to label and measure several biological markers simultaneously.

The unique optical properties of these semiconductor nanocrystals also hint at the possibility of observing changes that take place in labeled biological systems, such as living cells, over a period of time. This research was reported in the September 25, 1998 issue of the journal Science.

Alivisatos is a leader in the production by chemical means of semiconductor nanocrystals, simple inorganic solids consisting of a hundred to a hundred thousand atoms. He holds a joint appointment as a UCB professor of chemistry and a senior staff scientist with Berkeley Lab's Materials Sciences Division (MSD). Weiss, a staff scientist with MSD, is an authority on single molecule fluorescence and spectroscopy. He approached Alivisatos with the proposal for this joint study.

Other members of the team who co-authored the paper in Science were Marcel Bruchez Jr., who also holds a joint LBNL-UCB appointment, and Mario Moronne and Peter Gin, with Berkeley Lab's Life Sciences Division.

"Form follows function" is the golden rule in cell biology, which is why microscopy has been the heart and soul of this research field and fluorescent-labeling one of its most widely used tools. In fluorescent labeling, markers, usually antibodies that attach themselves to specific proteins, are tagged with dye-molecules that fluoresce or emit a specific color of light when stimulated by laser light, usually from a confocal microscope.

"Sometimes, in order to fully characterize a sample, a population of cells, for example, you need to look at combinations of markers," says Alivisatos. Such measurements require multiple-color light emissions which are difficult to obtain with conventional dye molecules.

"Ideal probes for multi-color experiments should emit at spectrally resolvable energies, should have a narrow, symmetric emission spectrum, and the whole family should be excitable at a single wavelength," the authors of the Science paper wrote.

Semiconductor nanocrystals met these demands in a "dual emission from single excitation" labeling experiment on mouse tissue cells called 3T3 fibroblasts. A core nanocrystal of cadmium selenide was enclosed within a shell of cadmium sulfide to boost the amount of fluorescence and reduce photochemical degradation. This core-shell combo was then enclosed within a shell of silica for water solubility and biocompatibility.

With earlier work by Alivisatos having shown that the color of light emitted by a semiconductor nanocrystal depends upon its size, the mouse cells were labeled with two different sizes of core-shell nanocrystals. It was also known that modifying the surface of the silica shell can be used to selectively control its attachment to components within a cell. In this case, the smaller nanocrystals (two nanometers), which fluoresced green, were modified to penetrate the nucleus of each cell, and the larger nanocrystals (four nanometers), which emitted red light, were modified so that they would attach themselves to actin filaments along the outer cell membrane.

Using wide-field microscopy, the green and red labels were clearly visible to the naked eye and could be photographed in true color with an ordinary camera. Confocal microscopy images showed that cell nuclei had been penetrated with the green probes and the actin fibers had been stained red. After repeated scans, the nanocrystal labels showed far less photobleaching than would occurred in the control sample labeled with conventional dye molecules.

"The development of semiconductor nanocrystals for biological labeling gives biologists an entire new class of fluorescent probes for which no small organic molecule equivalent exists," the authors of the Science paper wrote. "These nanocrystal probes can be complementary and in some cases may be superior to existing fluorophores."

The authors also assert that, compared with conventional fluorophores, semiconductor nanocrystals have a "narrow, tunable, symmetric emission spectrum, and are photochemically stable." These features, along with a relatively long fluorescence lifetime (hundreds of nanoseconds) indicate that, in addition to serving as direct probes, semiconductor nanocrystals could also be used as "sensitizers" for traditional dye-molecule probes, meaning they transfer their excitation energy to the dye-molecule.

An earlier research team led by Weiss demonstrated that this energy sensitizing phenomenon, known as fluorescence resonance energy transfer (FRET), when it takes place between a single donor and a single receptor, could allow for the labeling and observation of dynamic events such as conformational changes in a protein.

"The use of semiconductor nanocrystals should allow us to do unique FRET experiments," says Weiss. "For example, labeled molecules could be made to emit different colors at different times of an event."

Weiss and Alivisatos and their co-authors also believe that semiconductor nanocrystals could be applied to x-ray- and electron-based imaging techniques, and could serve as tunable infrared dyes for detecting fluorescence in, among other things, blood samples.

Berkeley Lab is a U.S. Department of Energy national laboratory located in Berkeley, California. It conducts unclassified scientific research and is managed by the University of California.


Story Source:

The above story is based on materials provided by Lawrence Berkeley National Laboratory. Note: Materials may be edited for content and length.


Cite This Page:

Lawrence Berkeley National Laboratory. "Semiconductor Nanocrystals: The Next Thing In Fluorescent Probes." ScienceDaily. ScienceDaily, 28 September 1998. <www.sciencedaily.com/releases/1998/09/980928072432.htm>.
Lawrence Berkeley National Laboratory. (1998, September 28). Semiconductor Nanocrystals: The Next Thing In Fluorescent Probes. ScienceDaily. Retrieved August 31, 2014 from www.sciencedaily.com/releases/1998/09/980928072432.htm
Lawrence Berkeley National Laboratory. "Semiconductor Nanocrystals: The Next Thing In Fluorescent Probes." ScienceDaily. www.sciencedaily.com/releases/1998/09/980928072432.htm (accessed August 31, 2014).

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