Reflective optical coating

NEXTERION® HiSens Slides

The majority of microarray users prefer to use fluorescent dyes as labels in their experiments, as these dyes typically offer high sensitivity and enable the detection of two or more different dyes simultaneously. Unfortunately, microarray scanners are not able to operate at their maximum efficiency, as the detectors do not acquire all 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 has utilized its extensive optical glass coating expertise to develop the next generation of microarray slides – NEXTERION® HiSens (High Sensitivity).


Overview

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 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

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

Immobilization chemistry

NEXTERION® HiSens Slides are available with most of the NEXTERION® functional coatings.
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.


Product details

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.
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.

Protocols

DNA Applications

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: March 2013
Protocol version: 3.0
Revision made: New phone number.
New composition of blocking solution.

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.

Protein Applications

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.



Publications citing NEXTERION® HiSens Slides


The complete publication list can be downloaded here!


Lampe, Paul, Hanash, Samir Fred Hutchinson Cancer Research Center
Affinity-based strategies to fast track development of colon cancer biomarkers
NIH Early Detection Research Network (EDRN) Protocol ID: 335
Full publication http://edrn.nci.nih.gov/protocols/335-affinity-based-strategies-to-fast-track

Troy Anderson, Julia Wulfkuhle, Emanuel Petricoin, and Raimond L. Winslow 2011
High resolution mapping of the cardiac transmural proteome using reverse phase protein microarrays
Mol Cell Proteomics mcp.M111.008037. First Published on April 13, 2011,
doi:10.1074/mcp.M111.008037
Full version http://www.mcponline.org/content/early/2011/04/13/mcp.M111.008037.full.pdf

Patrick Domnanich, Dacimoneida Brito Peña, Claudia Preininger
Xanthan/chitosan gold chip for metal enhanced protein biomarker detection
Biosens Bioelectron. 2011; 26(5):2559-2565
Full version: http://www.sciencedirect.com/science/article/pii/S0956566310007670

Stoevesandt O, Vetter M, Kastelic D, Palmer EA, He M, Taussig MJ
Cell free expression put on the spot: Advances in repeatable protein arraying from DNA (DAPA)
N Biotechnol. 2011; 28(3)#282-290
Full version: http://www.sciencedirect.com/science/article/pii/S1871678410005650

Denapaite D, Brückner R, Nuhn M, Reichmann P, Henrich B, Maurer P, Schähle Y, Selbmann P, Zimmermann W, Wambutt R, Hakenbeck R
The genome of Streptococcus mitis B6 - What is a commensal?
PLoS One. 2010; 5(2):e9426
DOI: 10.1371/journal.pone.0009426
Full version: http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0009426

Kontunen-Soppela S, Parviainen J, Ruhanen H, Brosché M, Keinänen M, Thakur RC, Kolehmainen M, Kangasjärvi J, Oksanen E, Karnosky DF, Vapaavuori E
Gene expression responses of paper birch (Betula papyrifera) to elevated CO2 and O3 during leaf maturation and senescence
Environ Pollution. 2010; 158(4):959-968
DOI: 10.1016/j.envpol.2009.10.008
Full version: http://www.sciencedirect.com/science/article/pii/S0269749109004886

Merkel O, Hamacher F, Laimer D, Sifft E, Trajanoski Z, Scheideler M, Egger G, Hassler MR, Thallinger C, Schmatz A, Turner SD, Greil R, Kenner L
Identification of differential and functionally active miRNAs in both anaplastic lymphoma kinase (ALK)+ and ALK− anaplastic large-cell lymphoma
Proc Natl Acad Sci U S A. 2010; 107(37):16228-16233
DOI: 10.1073/pnas.1009719107
Full version: http://www.pnas.org/content/107/37/16228

Sawle AD, Wit E, Whale G, Cossins AR
An information-rich, alternative, chemicals testing strategy using a high definition toxicogenomics and zebrafish (Danio rerio) embryos
Toxicol Sci. 2010; 118(1):128-139
DOI: 10.1093/toxsci/kfq237
Full version: http://toxsci.oxfordjournals.org/content/118/1/128

Spahn M, Kneitz S, Scholz CJ, Nico S, Rüdiger T, Ströbel P, Riedmiller H, Kneitz B
Expression of microRNA-221 is progressively reduced in aggressive prostate cancer and metastasis and predicts clinical recurrence
Int J Cancer. 2010; 127(2):394-403
DOI: 10.1002/ijc.24715
Full version: http://onlinelibrary.wiley.com/doi/10.1002/ijc.24715/abstract;jsessionid=DBBF123B6950516630FFFFD4E5181FFB.d02t03

Leberre V, Baranowski E, Deplanche M, Trouilh L, François JM
Detection of minority variants within bovine respiratory syncytial virus populations using oligonucleotide-based microarrays
J Virol Methods. 2008;148(1-2):271-276
DOI: 10.1016/j.jviromet.2007.10.026
Full version: http://www.sciencedirect.com/science/article/pii/S0166093407004363 Marino V, Galati C, Arnone C Optimization of fluorescence enhancement for silicon-based microarrays J Biomed Opt. 2008; 13(5):054060D DOI: 10.1117/1.2992142 Full version: http://spiedigitallibrary.org/jbo/resource/1/jbopfo/v13/i5/p054060_s1?isAuthorized=no

Son A, Nichkova M, Dosev D, Kennedy IM, Hristova KR
Luminescent lanthanide nanoparticles as labels in DNA microarrays for quantification of methyl tertiary butyl ether degrading bacteria
J Nanosci Nanotechnol. 2008; 8(5):2463-2467
DOI: 10.1166/jnn.2008.347
Full version: http://www.ingentaconnect.com/content/asp/jnn/2008/00000008/00000005/art00035?token=004912538c5c5f3b3b4746214874343e703a796d7b734f582a2f433e402c3568263c2b450

N.N.
miRNA Expression Profiling
Ambion TechNotes. Aug 2006;13(3):17
Full version: http://www.ambion.com/techlib/tn/133/AmbionTechNotes13_3.pdf

Redkar RJ, Schultz NA, Scheumann V, Burzio LA, Haines DE, Metwalli E, Becker O, Conzone SD
Signal and sensitivity enhancement through optical interference coating for DNA and protein microarray applications
J Biomol Tech. 2006; 17(2):122-130
Full version: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2291774/

Trade Fairs & Events
21.
March
Trade fair Workshop "Microarrays: Basics, Production and Processing", ETH Zurich / University of Zurich, Switzerland, 21.03 - 22.03.2017