Until now, however, the use of DNA microarray assays has been limited because current techniques typically depend upon fluorescence detection, a demanding methodology that requires time-consuming chemical labeling, high-power excitation sources, and sophisticated instrumentation for scanning. Such demands are generally well beyond the capabilities of individual laboratories or clinics, especially in developing countries. While label-free DNA detection strategies do exist, they require either complex device fabrication or sophisticated instrumentation for readouts, and in addition none are compatible with conventional DNA microarrays, where up to one million sequences are available for interrogation in a single experiment.
"We have demonstrated parallel sampling of a microarray surface with micron-scale resolutions over centimeter-scale lengths," said Groves. "This is four orders of magnitude larger than what has been achieved to date with conventional scanning-electrostatic-force microscopy."
In a typical experiment, a microarray is prepared and mounted in a well chamber and the DNA is hybridized (a standard technique in which complementary single strands of DNA bind to form double-stranded DNA "hybrids"). A suspension of negatively-charged silica microspheres is then dispersed through gravitational sedimentation over the microarray surface, a process which takes about 20 minutes. Because the substrate or background surface of the microassay is positively charged, the silica microspheres will spread across the entire surface and adhere to it. However, on surface areas containing double-stranded DNA, which is highly negatively charged, and on areas containing
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| Contact: Lynn Yarris lcyarris@lbl.gov 510-486-5375 DOE/Lawrence Berkeley National Laboratory Source:Eurekalert |