The QIAGEN HiLight System uses Resonance Light Scattering (RLS) instead of fluorescence for detection. The intense signals produced by the HiLight System enable simple and highly sensitive detection of nucleic acids without the need for amplification steps. The HiLight System is part of QIAGEN¡¯s complete range of products for highly sensitive and reliable microarray applications.
The QIAGEN HiLight Array Detection System uses Resonance Light Scattering (RLS), a technology based on the optical properties unique to metal colloids of defined size, shape, and composition. Light scattering is the process by which energy from an incident beam of light is re-radiated as scattered light. The energy from the incident light can be both absorbed and scattered. HiLight RLS Particles SC exposed to light at a wavelength far removed from their absorption range, however, can act as "pure" light scatterers, which scatter the light but do not absorb energy. Because of the metallic nature of the particles, there is no reduction of the signal due to photochemical bleaching. The stable, archivable signal allows multiple reads to detect low- and high-abundance mRNAs under optimal conditions and to standardize the signal between assays or between different HiLight Readers. HiLight technology eliminates the long exposure times needed for methods that use radioactivity while providing high sensitivity.
The HiLight Reader and the HiLight Kits form a fully integrated microarray detection system that enables the user to perform nucleic acid expression experiments on functionalized glass surfaces. The detection system uses biotinylated target cDNA and anti-biotin-coated gold particles (see figure "HiLight Principle"). HiLight products are designed to provide greater sensitivity and to improve the lower limit of detection compared with current fluorescence technologies.
HiLight Principle
Schematic showing labeling and detection of biotin-labeled cDNA using anti-biotin coated HiLight RLS Particles SC.
The HiLight System works with both commercial and self-spotted slides. The procedure uses biotinylated target cDNA which is hybridized with the immobilized capture probe on the microarray. Following a post-hybridization wash and a short blocking step, anti-biotin- coated gold particles are used for the highly sensitive detection of differentially expressed genes. An optical coating is applied to assure high signal- to- background ratios and to protect the slides from environmental hazards.
HiLight Kits provide optimized HiLight RLS Particles SC and hybridization, washing, blocking, and staining reagents, with protocols to perform experiments on 5 or 24 separate microarray slides.
The HiLight Single-Color Kit Plus provides the additional LabelStar Array Kit for highly efficient labeling and cleanup guaranteeing optimal labeling conditions for HiLight detection. The optional HiLight Accessory Kit contains accessories to facilitate array processing using the HiLight protocol. Detection is performed on the HiLight Reader. The HiLight Reader Plus provides a customized image analysis option using ArrayVisionRLS software.
The HiLight System uses a new detection technology for gene expression analysis to improve the sensitivity of your existing array platform. The increased signal intensity allows specifically high sensitivity applications such as:
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