NEXTERION® HiSens Slides are available with most of the NEXTERION® functional coatings.
The NEXTERION® HiSens product is an innovative microarray slide based on an ultraflat, high quality borosilicate glass slide coated with reflective dielectric layers (US patent No. 7939339 and European patent No. EP1591773). The reflective layers on NEXTERION®HiSens significantly enhance sensitivity and signal response. The characteristics of the reflective layers have been optimized for the fluorescent wavelengths most commonly used in microarray experiments, and will simultaneously improve the performance in both the Cy3™ and Cy5™ channels. The slide is produced according to industry standard slide dimensions and is available with SCHOTT’s standard high quality functional coatings for DNA and protein microarraying. This means that the NEXTERION® HiSens Slide is fully compatible with all microarray printing technologies and most slide processing protocols, allowing customers a smooth transition from industry-standard transparent microarray slides.
The NEXTERION® HiSens coating is able to enhance signal amplification by exploiting several optical effects:
- Fluorescence enhancement and reflection of excitation light
On a standard glass microarray slide, the fluorescently labelled spots absorb only some of the excitation light. By placing a dielectric coating beneath the spots, this “lost” light is reflected back onto the spots. In addition, the multiple layers of the HiSens coating have been specifically designed so that the incoming and reflected excitation light waves overlap and interfere with each other. The interference is constructive at the slide surface, enhancing the excitation by a theoretical factor of up to 6.3 times.
- Reflection of the emission light.
- The fluorescence emission in the Cy3™ and Cy5™ channels from the labelled spots is redirected towards the detector rather than being lost by passing through the glass or by scattering. This effectively doubles the detected fluorescence signal.
Taken together, these effects can theoretically enhance the fluorescence signal by up to 12.6 times when compared to standard transparent microarray slides.
Increased sensitivity
Significant increases in fluorescent signal intensities and signal-to-background ratios have been demonstrated with the NEXTERION® HiSens E and A+ products, the first two slide surfaces launched by SCHOTT in the HiSens range in 2006 (all slide surfaces are now available). The functional coatings are identical to the standard NEXTERION® slides and were processed with standard slide processing protocols. The results clearly exhibit the same impressive spot size and morphology that users typically associate with NEXTERION® Slide E and Slide A+, but the signal intensity is significantly enhanced thanks to the HiSens coating.
Identical oligonucleotide probes were printed on NEXTERION® HiSens A+ and the standard NEXTERION® Slide A+. The slides were hybridized and scanned on an Axon 4000B scanner with identical settings. Images of the NEXTERION® HiSens reflective coating are compared to standard Slide A+. The NEXTERION® HiSens A+ data was normalized against the standard NEXTERION® Slide A+. An 8 to 10 fold increase in signal-to-background ratios was obtained with the NEXTERION®HiSens reflective coating compared to the standard NEXTERION® Slide A+.
Improved signal response
NEXTERION® HiSens is the ultimate microarray slide for microarray applications where the target material is at a low concentration or cannot be reliably amplified, for example with mRNAs or low abundant proteins. The NEXTERION® HiSens coating may be used to produce reproducible signals at a much lower target concentration than with conventional slides.
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The graph shows that for a DNA microarray application, between two and ten fold less target was required to produce the same signal intensity with NEXTERION® HiSens as with a conventional aminosilane microarray slide, resulting in potentially significant cost savings for users.
Preservation of dye ratios
The reflective layers in the NEXTERION® HiSens coating have been designed to enhance the performance of both the Cy3™ and Cy5™ channels equally. The Cy3™ to Cy5™ ratios are preserved, allowing use of standard data normalization methods and direct comparison with data previously produced on conventional slides.