Lab Use of Qdots Specified

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In a step toward earlier disease diagnosis and personalized medicine, detailed protocols have been developed for using bioconjugated quantum dots -- luminescent nanoparticles linked to biological molecules -- to track biomarkers in cells and tissues.

Using prostate cancer specimens, researchers at Emory University and the Georgia Institute of Technology have confirmed that bioconjugated quantum dots (qdots) are effective in simultaneously identifying multiple molecular biomarkers in cancer tissue. The qdots have shown great promise as tools for disease diagnosis and treatment, but until now their medical use has been limited by the lack of specific instructions for clinicians.

The new protocols detail how to prepare, process and quantify these tiny particles, allowing them to be used as biomarkers. The technology is a variation of immunohistochemistry, the laboratory staining process commonly used by pathologists to identify proteins in a tissue section from a cancer patient.
Shuming Nie, a distinguished professor in the Wallace H. Coulter Department of Biomedical Engineering at Georgia Institute of Technology and Emory University, in the laboratory. Nie and colleagues have developed detailed protocols for using bioconjugated quantum dots to track molecular biomarkers in cells and tissues, a step toward earlier disease diagnosis and personalized medicine. (Photo courtesy Georgia Institute of Technology)
The scientists developed detailed protocols for using the technology, including antibody conjugation, preparation of tissue specimens, multicolor quantum staining, image processing and biomarker quantification. They also have developed bioinformatics and software tools for automated feature extraction and biomarker quantification.

The Emory-Georgia Tech team was led by Shuming Nie, a distinguished professor in the Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University, and by May Dongmei Wang, assistant professor in the Coulter Department and Georgia Cancer Coalition Distinguished Scholar.

"We have now resolved a major bottleneck in the use of multicolor quantum-dot probes for cancer immunohistostaining,” said Nie. “Quantum dot probes used in tissue diagnosis are considered to be one of the most important and clinically relevant applications for cancer technology in the near term. We believe that this technology will be ultimately useful in correlating a panel of biomarkers with disease progression and therapeutic response.”

“Personalized medicine is poised to transform health care over the next several decades,” said Wang, director of the bioinformatics and biocomputing core in the Emory-Georgia Tech Nanotechnology Center. “New diagnostic and prognostic tools will increase our ability to predict the likely outcomes of drug therapy. Essential to this endeavor is the use of bioinformatics and systems biology to link each individual’s molecule profile with disease diagnosis and treatment decisions.”

Nanoparticles, which can be as tiny as 100,000 times smaller than the width of a human hair, have special “quantum” properties, including changes in color according to minute differences in size. Bioconjugated qdots are collections of different-sized nanoparticles embedded in tiny beads made of polymer material. In a process called “multiplexing,” they can be finely tuned to a myriad of lumninescent colors that can tag a multitude of different protein biomarkers or genetic sequences in cells or tissues.

Because the qdots have a cadmium core, scientists have been concerned about their potential toxicity if infused into the bloodstream of patients. Using them in the laboratory to detect biomarkers in cells and tissues outside the human body eliminates this concern. Qdots also have advantages over traditional dyes and stains often used in imaging. They are more brightly fluorescent, they resist photobleaching and they can emit a broad range of colors simultaneously.

These properties make bioconjugated qdots highly promising for improving the sensitivity of disease diagnosis in the laboratory, and they are particularly important for detecting and analyzing cancer biomarkers that are present at low concentrations or in small numbers of cells. Biomarkers include altered or mutant genes, RNA, proteins, lipids, carbohydrates and small metabolite molecules.

"Aggressive cancer behaviors also could be better understood and rapidly predicted using these kinds of biomarkers,” said Nie. “By defining the interrelationships between biomarkers, it could be possible to diagnose and determine cancer prognosis based on a patient’s molecular profile, leading to personalized and predictive medicine.”

The Emory and Georgia Tech scientists also believe the bioconjugated qdots will be useful in detection of bioterrorism agents such as anthrax, plague, botulism and viral hemorrhagic fevers.

Wang’s group at Georgia Tech has developed a number of software tools for cancer nanotechnology, leading a major effort in “bio-nano-info” integration for personalized medicine.

The research was supported by a Bioengineering Research Partnership award and a Centers of Cancer Nanotechnology Excellence (CCNE) award, both from the National Cancer Institute. The work was also supported by the Microsoft eScience Program, the Georgia Cancer Coalition (GCC) and the Georgia Research Alliance (GRA).

Key faculty investigators at Emory included Leland Chung, professor of urology; Dr. Ruth O’Regan, associate professor of hematology and oncology and Georgia Cancer Coalition Distinguished Scholar; Dr. John Petros, professor of urology; and Dr. Jonathan Simons professor of biomedical engineering, hematology and oncology.

The research guidelines and results were published in the May 3 issue of Nature Protocols.

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Published: May 2007
A biomarker, short for biological marker, refers to a measurable and quantifiable indicator of a biological condition, process, or response. Biomarkers can be substances or characteristics that are objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes, or pharmacological responses to therapeutic interventions. These markers are often used in medical and scientific contexts to assess health status, diagnose diseases, monitor treatment outcomes,...
Immunohistochemistry (IHC) is a method for detecting antigens or haptens in cells of a tissue section by using labeled antibodies to bind specifically to their antigens. IHC is used for disease diagnosis, biological research and in drug development. Using IHC visualized tumor markers such as enzymes, oncogenes and tumor-specific antigens, physicians can diagnose if a tumor is benign or malignant and determine the stage and grade of a cancer.
The combination of two or more signals for transmission along a single wire, path or carrier. In most optical communication systems this is referred to as wavelength division multiplexing, in which the combination of different signals for transmission are imbedded in multiple wavelengths over a single optical channel. The optical channel is a fiber optic cable or any other standard optical waveguide.
An SI prefix meaning one billionth (10-9). Nano can also be used to indicate the study of atoms, molecules and other structures and particles on the nanometer scale. Nano-optics (also referred to as nanophotonics), for example, is the study of how light and light-matter interactions behave on the nanometer scale. See nanophotonics.
A small object that behaves as a whole unit or entity in terms of it's transport and it's properties, as opposed to an individual molecule which on it's own is not considered a nanoparticle.. Nanoparticles range between 100 and 2500 nanometers in diameter.
The technology of generating and harnessing light and other forms of radiant energy whose quantum unit is the photon. The science includes light emission, transmission, deflection, amplification and detection by optical components and instruments, lasers and other light sources, fiber optics, electro-optical instrumentation, related hardware and electronics, and sophisticated systems. The range of applications of photonics extends from energy generation to detection to communications and...
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