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Reflective optical coating
Nexterion® HiSens Slides
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The majority of microarray users prefer to use fluorescent dyes as labels in
their experiments, as these dyes typically offer the most sensitivity and also
enable detection of two or more different dyes simultaneously. Unfortunately,
microarray scanners do not operate at their maximum efficiency with fluorescent
dyes, as the detectors only acquire a small proportion of the total emitted
fluorescence.
There are a number of factors that contribute to the inefficient signal
capture:
- Only a proportion of the total excitation light is absorbed by the micro array
spots, with most passing through the glass slide.
- On standard non-reflective slides, there is weak but “destructive” interference
of the excitation light waves, leading to a lower fluorescent emission.
- The light emitted by the fluorescent dyes attached to the target molecule is
undirected and lost through the back of the transparent glass.
To address these problems, SCHOTT utilized its extensive optical glass coating
expertise to develop the next generation of microarray slides – Nexterion®
HiSens (High Sensitivity).
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| Type of
coating |
Ordering information |
| Nexterion® product |
Barcode option |
Item number |
Slides per pack |
| Reflective optical coating
available with most of Nexterion®
functional coatings |
HiSens uncoated |
None |
1141518 |
25 |
| Label |
1141519 |
25 |
| HiSens AStar |
None |
1178045 |
25 |
| Label |
1178046 |
25 |
| HiSens A+ |
None |
1139673 |
25 |
| Label |
1141515 |
25 |
| HiSens AL |
None |
1139827 |
25 |
| Label |
1141514 |
25 |
| HiSens E |
None |
1125813 |
25 |
| Label |
1137332 |
25 |
| HiSens H |
None |
1141517 |
25 |
| Label |
1139829 |
25 |
| HiSens P |
None |
1178051 |
25 |
| Label |
1178052 |
25 |
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| Key product features |
| • Significantly improved signal to background ratios for identifying
low-expressor genes or low-abundant proteins |
| • Easy implementation with standard slide protocols |
| • Compatible with all common microarray equipment |
| • Reduced target concentrations |
| • Reduced reagents and dye costs |
| • Preservation of Cy3/5™ ratios – comparable to data from conventional
slides |
| • Chemically stable reflective coating |
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Nexterion® HiSens Slides are available with most of the Nexterion® functional
coatings.
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| Suitable probe types |
| The functional coating on Nexterion® HiSens Slides is identical to the
respective coating on Nexterion® Slides. Please refer to the table in the
section “General information coated slides” in order to find the appropriate
coating chemistry for your specific application. |
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Product details
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The Nexterion® HiSens product is an innovative microarray slide based on an
ultraflat, high quality borosilicate glass slide coated with reflective dielectric
layers (patent pending). 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.
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- 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.
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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.
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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+.
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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.
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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.
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Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
Protocol revision
Date: April 2009
Protocol version: 2.0
Revision made: New method for identifying the printing side.
New telephone and fax numbers.
Revision reason: New coating technology for reflective coating.
Update contact information.
|
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Contact
Microarray Solutions
SCHOTT North America Inc.
5530 Shepherdsville Road
Louisville, KY 40228
USA
 | +1 -502-657-4411 |
 | +1 -502-966-4976 |
E-mail
More contacts
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