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6.29.1 Sensor for Detecting Minute Quantities of Biological Materials
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A new device based on laser-excited fluorescence provides unparalleled detection of biological materials for which only minuscule samples may be available. This device, invented at the Ames Laboratories, received a 1991 R&D 100 award. The Ames Microfluor detector was developed to meet a need for an improved detection technique, driven by important new studies of the human genome, abuse substances, toxins, DNA adduct formation, and amino acids, all of which may be available only in minute amounts. Although powerful and efficient methods have been developed for separating biological mixtures in small volumes (i.e., capillary electrophoresis), equally powerful techniques for subsequent detection and identification of these mixtures have been lacking. The Microfluor detector combines very high sensitivities with the ability to analyze very small volumes. The instrument design is based on the principle that many important biomaterials are fluorescent, while many other biomaterials, such as peptides and oligonucleotides, can be made to fluoresce by adding a fluorescent tag. When a sample-filled capillary tube is inserted into the Microfluor detector and is irradiated by a laser beam, the sample will fluoresce. The detector detects, monitors, and quantifies the contents by sensing the intensity of the fluorescent light emitted. The signal is proportional to the concentration of the materials. The proportionality constant is characteristic of the material itself. Analyses can be performed with sample sizes 50 times smaller than those required by other methods, and concentrations as low as 10 11 molar (1 part per trillion) can be measured. Often, the critical components in a sample are present at these minute concentrations. These two features make the Microfluor detector uniquely compatible with capillary electrophoresis. In addition, the Ames-developed detector is distinct from other laser-excited detectors in that it is not
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seriously affected by stray light from the laser itself; it also allows simple alignment and operation in full room light. The Microfluor detector has already been used to determine the extent of adduct formation and base modification in DNA so that the effects of carcinogens on living cells can be studied. Future uses of the sensor will include DNA sequencing and protein sequencing. With direct or indirect fluorescence detection, researchers are using this technique to study the chemical contents of individual living cells. This capability may allow pharmaceutical products to be tested on single cells rather than a whole organism, with improved speed and safety.
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6.29.2 Sensors for Early Detection and Treatment of Lung Tumors
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A quick accurate method for early detection of lung cancer would raise chances of patient survival from less than 50 percent to 80 or 90 percent. Until now, small cancer cells deep in the lung have been impossible to detect before they form tumors large enough to show up in x-rays. Researchers at Los Alamos National Laboratory in collaboration with other institutions, including the Johns Hopkins school of medicine and St. Mary s Hospital in Grand Junction, Colorado, are developing methods for finding and treating lung cancer in its earliest stages. A detection sensor involves a porphyrin, one of an unusual family of chemicals that is found naturally in the body and concentrates in cancer cells. The chemical is added to a sample of sputum coughed up from the lung. When exposed to ultraviolet or laser light, cells in the porphyrin-treated sputum glow a bright red. When the sample is viewed under the microscope, the amount and intensity of fluorescence in the cells determines the presence of cancer. The first clinical test of a detection technique using porphyrin was done by LANL and St. Mary s Hospital in 1988. Four different porphyrins were tested on sputum samples from two former miners, one known to have lung cancer and one with no detectable cancer. One of the porphyrins was concentrated in certain cells only in the sputum of the miner with lung cancer. Other tests concluded that these were cancer cells. Later, a blind study of sputum samples from 12 patients, 8 of whom had lung cancer in various stages of development, identified all the cancer patients as well as a ninth originally thought to be free of cancer. Further tests showed that this patient also had lung cancer. Identifying the ninth patient prompted a new study in which the procedure was evaluated for its ability to detect precancerous cells in the lung. In this study, historical sputum samples obtained from Johns Hopkins were treated with the porphyrin. Precancerous conditions that chest x-rays had not detected were identified in samples from patients who later developed the disease. Although further
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