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Multi-well formats
General
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Multi-well microarray substrates allow the parallel analysis of multiple biological
samples against focused subsets of probes. The multi-well format permits
a number of versatile assay designs such as multiplexed experiments, sideby-
side comparisons, or replicate experiments to be performed. Additional
advantages of using multiple well formats include higher array-to-array
reproducibility and lower experimental costs when compared to conventional
single-array substrates. SCHOTT can offer multi-well coated substrates in two
formats: Nexterion® MPX-16-wells on a 75.6 mm x 25.0 mm glass slide, or
the Nexterion® MTP-96-wells in microplate format. The MPX-16 slide is fully
compatible with standard microarray slide printers and scanners, whereas the
MTP-96 conforms to the SBS standard format commonly used in clinical diagnostics
and drug discovery. The multi-well formats are available with functional
chemistries and coatings that are identical to the standard Nexterion®
slides, allowing assays to be easily transferred to the high throughput multiwell
formats.
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| Formats |
Immobilization
method |
Typical applications |
16-well slide
96-well SBS microplate |
Various |
• Surveillance chips
• Virus or bacterial identification
• Molecular diagnostics
• Dose response studies |
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| Key product features |
| • Simultaneous analysis of multiple biological samples in one microarray
experiment |
| • Excellent reproducibility compared to conventional single-array
applications |
| • Optimal use of limited sample material |
| • Substantial reductions in cost per experiment |
| • Compatible with standard commercial printing, liquid handling and
scanning equipment |
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| Nexterion®
coating |
Type |
Nexterion® MPX-16
Item number |
Nexterion® MTP-96
Availability |
| None |
Uncoated glass |
1079386 |
Also available without 96-well
patterning |
| AStar |
Aminosilane |
1178047 |
available |
| A+ |
Aminosilane |
1078353 |
available |
| AL |
Aldehydesilane |
1078355 |
available |
| E |
Epoxysilane |
1078354 |
Also available without 96-well
patterning |
| H |
3-D thin film polymer with NHS-ester |
1091524 |
Not available |
| P |
3-D thin film polymer with NHS-ester |
1178053 |
Under development |
| NC-W |
White nitrocellulose |
1178059 |
Under development |
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Product details
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Hydrophobic multi-well patterning
The microarray quality glass substrates are printed with either a 16- or 96-
well hydrophobic patterning, and are available with a number of functional
coatings, making the both formats suitable for most microarray applications
using DNA, proteins, or other bio-molecules as probes. The black patterning
material serves several important functions, one of which is to create the individual
wells that act as a hydrophobic barrier preventing cross contamination
between the arrays. The well pattern also serves as a registration aid for probe
deposition, and a reproducible surface for the adhesion of the silicone superstructure.
The absence of well rims makes Nexterion® multi-well substrates
compatible with all microplate arrayers and significantly reduces print times
compared to substrates that have recessed wells. The patterned glass substrates
have a corner indicator to provide an unambiguous orientation mark.
Most modern robotic arrayers are equipped to handle the 96-well microplate
formats, and many have pre-programmed definitions for both the Nexterion®
MPX-16 slides and Nexterion® MTP-96 plates, simplifying the set up process
for users. Information on compatible arrayers and detailed instructions for
printing into the Nexterion® multi-well formats are available on request.
Avoidance of cross-contamination
Cross contamination is avoided during multiplexed hybridization as a result
of the 16- or 96-well hydrophobic patterning, and the use of the optional
self-adhesive silicone superstructures.
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The figure shows a pictorial representation of subarray
wells from Nexterion® Slide MPX-16 that were
first printed with three distinct nucleic acid probes
and then hybridized with various combinations of
three complementary, fluorescent-labeled targets.
No cross contamination was observed.
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Intra-slide reproducibility
Nexterion® multi-well coated substrates allows users to obtain microarray data
with exceptionally high reproducibility. This was demonstrated by hybridizing
multiple 10 x 10 DNA arrays with fluorescent-labeled cDNA targets using
Nexterion® Slide MPX-16. Correlation coefficients were obtained by comparing
the normalized signal intensity from spots in different subarray wells. An
average correlation coefficient of 0.96 was obtained after pair wise comparison
of all eight-subarray wells.
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Virtually identical subarrays in each well of one
Nexterion® Slide MPX-16.
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Inter-slide reproducibility
The excellent inter-slide reproducibility offered by Nexterion® multi-well
coated substrates is attributable to the simultaneous analysis of all subarray
assays, and to the improved sample mixing during hybridization within the
wells of the superstructure. The figure shows the average correlation coefficient
(R2) obtained with Nexterion® Slide MPX-16 (indicated as MPX) and
conventional slides (indicated as Non-MPX). Experiments were conducted as
described in the previous figure, but now using multiple slides for same day
and day-to-day experiments.
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Optimal use of limited target material
The Nexterion® multi-well coated substrates allow lower target solution
volumes to be used compared to conventional slides. Identical probe sets
in separate subarray wells of Nexterion® Slide MPX-16 were hybridized with
7.5 pmol of fluorescent-labeled target (house-keeping gene: GAPDH), which
was dissolved in different volumes (60, 45, 30 µL) of hybridization solution.
The figure demonstrates that a reduction in target volume results in increased
target concentration and thus in improved signal intensities. Nexterion®
Slide MPX also offers cost savings by reducing the amount of target material
required without any loss of sensitivity (assuming that the target volume and
target amount are reduced equally, keeping the concentration consistent).
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Contact
Microarray Solutions
SCHOTT North America Inc.
5530 Shepherdsville Road Louisville, KY 40228 USA
 | +1 -502-657-4431 |
 | +1 -502-966-4976 |
E-mail
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