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testing is needed, researchers now feel confident that the technique and the detection sensor can detect precancerous conditions three to four years before onset of the disease. Working in collaboration with industry, researchers expect to develop an instrument in several years for rapid automated computerized screening of larger populations of smokers, miners, and other people at risk of developing lung cancer. Because lung cancer is the leading cancer killer in both men and women, a successful screening procedure would dramatically lower the nation s mortality rate from the disease. A similar procedure has potential for the analysis of Pap smears, now done by technicians who screen slides one at a time and can misread them. In addition to a successful screening program, LANL hopes to develop an effective treatment for early lung cancer. The National Cancer Institute will help investigate and implement both diagnostic and therapeutic programs. Researchers are performing basic cell studies on animals to investigate the effect of porphyrin on lung cancer cells. They are also exploring the use of the LANL-designed porphyrin attached to radioactive copper to kill early cancer cells in the lung in a single search-and-destroy mission. The porphyrin not only seeks out cancer cells but also has a molecular structure, similar to a hollow golf ball, that can take certain metals into its center. A small amount of radioactive copper 67 placed inside the porphyrin should destroy tumors the size of pinhead, as well as function as a tracer.
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Ultrasensitive Sensors for Single-Molecule Detection
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Enhancing the sensitivity of research instruments has long been a goal of physicists, biologists, and chemists. With the single-molecule detector, researchers have achieved the ultimate in this pursuit: detection of a single fluorescent molecule in a liquid. The new instrument a thousand times more sensitive than existing commercial detectors brings new capabilities to areas of science and technology that affect lives in many ways, from DNA sequencing, biochemical analysis, and virus studies to identifying environmental pollutants. For some time, scientists have observed individual molecules trapped in a vacuum, where they are isolated and relatively easy to find. However, many important biological and chemical processes occur in a liquid environment, where many billions of other molecules surround the molecules of interest. Developing a single-molecule detector that operates in such an environment presented a difficult challenge. The observation technique involves attaching fluorescent dye molecules to the molecules of interest and then exciting the dye molecule by passing them in solution to a rapidly pulsed laser beam and detecting the subsequent faint light, or photons, they emit. The fluorescent lifetimes of the molecules are much shorter than the time the
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FIGURE 6.31 solution.
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molecules spend in the laser beam; therefore each molecule is reexcited many times and yields many fluorescent photons. The signature of the passing molecule is the burst of photons that occurs when the molecule is passing the laser beam (Fig. 6.31). A lens, or microscope objective, and a slit are arranged to image the photons from a small region around the laser beam waist onto a microchannel plate photomultiplier (MCPP) that counts individual photons. The intense excitation light from the laser is blocked from reaching the MCPP by a bandpass spectral filter, which is centered near the peak fluorescent wavelength. The excitation light consists of extremely short pulses, each about 70 trillionths of a second. The dye molecule does not emit light until a billionth of a second after excitation, so the flash of laser light fades before the feeble molecular glow occurs. For reliable identification of individual molecules, the technique maximizes the number of the detected photons and minimizes the number of background photons. Although some background remains, the technique registers over 85 percent of the fluorescent molecules. Developed to aid in sequencing chromosomes for the Human Genome Project, the sensor detector s high activity would allow DNA sequencing rates hundreds of times faster than those obtainable with present techniques. It would also eliminate the need for radioactive materials, gels, and electrophoresis solutions, which often create disposal problems, and it is expected to help make DNA sequencing a routine diagnostic and clinical tool. One eventual benefit of the DNA research may be rapid screening for any of 3500 known genetic diseases such as diabetes, cystic fibrosis, and Alzheimer s disease. The ultrasensitive detector can be used to find and quantify minute amounts of chemicals, enzymes, and viruses in the blood and monitor the dispersal of extremely low concentrations of environmental pollutants. The device may also be useful in studying the interaction of viruses and their binding sites. It may be possible to
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