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Arrayjet Microarray Technology - An Assessment of Printing Reproducibility
Arrayjet Microarray Technology - An Assessment of Printing Reproducibility
Analysis of the Printing Consistency of Protein Microarrays
Using Arrayjet Marathon Microarrayers
Analysis of the Printing Consistency of Protein Microarrays Using Arrayjet Marathon Microarrayers
Introduction
This application note provides evidence of feature
reproducibility and consistency across slide batches
printed using Arrayjet Marathon microarrayers. It
highlights the successful batch to batch printing of
fluorescent tagged Bovine Serum Albumin (BSA) onto
Schott Nexterion® epoxysilane slides to produce high
quality spots.
Experimental Design
Sample preparation
Bovine Serum Albumin (BSA-IgG free, 1mg/mL,
Sigma) was diluted in a 47% glycerol buffer
containing 1µg/mL Rhodamine B. 20μL of sample
volume was deposited in 36 wells of a 384 well
microplate.
Substrates
Arrays were printed onto Schott Nexterion®
Epoxysilane slides.
Inkjet Printing
Each drop dispensed by an Arrayjet Marathon
microarrayer is 100pL. One drop per spot was
printed per sample, in triplicate across 48
miniarrays. To print a batch of 100 slides, 3
sample redraws were performed with 1.3μl of
sample aspirated each time .The temperature and
humidity (RH) were maintained between the
ranges 15-20°C and 40-60% RH respectively.
Image Acquisition and Analysis
Slides at different tray positions were scanned
using GenePix® 4000B scanner (Molecular
Devices). Images were acquired under a
wavelength of 532nm with 100% power gain and a
PMT of 230.
Data Analysis
Data was acquired with GenePix® Pro 6.0 4000B.
Mean intensity values of the median F 532 (minus
background) of all replicate spots were used for
signal calculation. The mean diameter of all
features was recorded.
Results
Intra batch analysis
The printing reproducibility of the Arrayjet
Marathon microarrayer was demonstrated within
one batch of 100 slides. A good spot morphology
was observed across all slides with consistent spot
positioning (Figure 1). The intraslide %CV values
(Table 1) were observed to be low thereby
indicating minimum variability within a single
slide. The mean intrabatch CV value was
calculated to be 4.14% representing consistent
signal intensity from all spots printed within a 100
slide batch. The spot size within a batch was in
range of 110μm to 115μm.
Inter batch analysis
Two batches of 100 slides were printed on the
same day and then again over two separate days
to demonstrate inter batch printing consistency
over time. Table 1 represents the results obtained
following data acquisition and analysis. The mean
signal intensity obtained from 2 separate batches
over two different days showed a low %CV value
of 4.36%, indicating consistency in print quality
and inter batch reproducibility over time. The
mean CV value for printing performed on the
same day was 5.08% demonstrating printing
reproducibility between sequential batch runs.
The spot size throughout all batches was in the
range of 108μm to 113μm.
Print Head Nozzle Performance
Arrayjet printing technology makes use of the
Xaar™ print head to deposit samples onto
substrates. A low volume 12 JetSpyder docks with
the print head to simultaneously load 12
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samples into different nozzle sets of the print head
(Figures 2A and B). The individual nozzle
performance was analysed by calculating the
mean %CV of all spots printed with the same
nozzle number (Figure 3). The mean %CV values
from different nozzles were calculated to be in the
range of 3- 4% highlighting spot precision and
printing accuracy using Arrayjet microarrayers.
Conclusion
Results obtained following intra batch and inter
batch slide analysis represent good printing
reproducibility (~5% CV values) and spot size
consistency using Arrayjet Marathon
microarrayers. When a protein sample such as
BSA was printed onto Schott Nexterion®
epoxysilane slides, high quality reproducible spots
were produced with minimum spot to spot, slide
to slide and batch to batch variability.
Furthermore, the accurate nozzle performance of
the print head combined with rapid on the fly
printing makes Arrayjet non-contact technology
ideal for manufacturing multiple slide batches
over time.
Figure 1: Representative image showing 3 replicates of BSA
sample across 4 miniarrays. Good spot morphology and
consistent spot positioning can be observed across all arrays.
References
McWilliam, I., Chong Kwan, M., and Hall, D.
(2011).Inkjet Printing for the Production of Protein
Microarrays. In: Protein Microarrays: Methods and
Protocols. (U. Korf, ed) Humana Press, New York
A
Inkjet print head
B
Samples introduced
into the print head
Jetspyder and print
head docking
Arrayjet
Jetspyder
Capillaries of the
Jetspyder
Print head
moving across
the slide in a
non-contact
fashion
depositing
samples as it
travels
Microtitre
plate
Figure 2A and B: Arrayjet print head and Jetspyder operation.
,The
p Jetspyder
o Inkjet
technology
is docked
to the print head, and moved to the wells of the microplate containing samples to be arrayed. 12
samples are simultaneously drawn through the Jetspyder into the nozzles of the print head. The nozzles of the print head
contain the samples to be printed. As the print head travels in a non-contact fashion across the slide, the samples get
deposited in the form of spots on the slide.
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Table
1: Intra
batch
andand
inter
batch
slide
analysis.
Table
1: Intra
batch
inter
batch
slide
analysis.
Arrayjet
Marathon
microarrayer
demonstrating
printing
reproducibility
and
spot
size
consistency.
Arrayjet
Marathon
microarrayer
demonstrating
printing
reproducibility
and
spot
size
consistency.
Table
1:Intensity
Intra batch
and inter
batch
slide analysis.
(Mean
=
median
532background
532)
(Mean
Intensity
=
median
532background
532)
Arrayjet Marathon microarrayer demonstrating printing reproducibility and spot size consistency.
(Mean Intensity = median 532- background 532)
Batch
1 1
Batch
Batch 1
Batch
1 1
Batch
Batch 1
Batch
1 1
Batch
Slide
position
Slide
position
2525
50
100
50position7575
100
Slide
Mean
Mean
Intra
slide
%CV
Intra 50
slide %CV 75
25
100
Intensity
Intensity
Mean
5.80
5.34
5.67
5.80 Intra slide
5.34
6.13
5.67
%CV 6.13
Intensity
slide
Diameter
(μm)
Intra
slide
Diameter
(μm)
5.80Intra
5.34
6.13
5.67
113.01
110.89
113.01
110.89 115.97
115.97 112.76
112.76
Intra slide
Diameter
(μm)
Intra
batch
Analysis
Intra batch112.76
Analysis
113.01
110.89
115.97
20509.18
21693.65
19704.73
21184.50
20509.18
21693.65 Intra
19704.73
21184.50 20773.02
20773.02
batch Analysis
Batch
1 1
Batch
Batch
2
Batch
Batch
1 2
Batch 2
Batch
1 1
Batch
Batch
2
Batch
1 2
Batch 2
%CV
(Sdev)
Mean
Mean
Diameter
Diameter
Mean
(μm)
(μm)
Diameter
(μm)
861.72
861.72
4.14
4.14
113.15
113.15
861.72
4.14
113.15
892.72
892.72
4.36
4.36
113.15
113.15
108.81
108.81
113.15
892.72
4.36
974.92
974.92
5.08
5.08
17679.36
17679.36 19192.64
19192.64 19262.74
19262.74 19907.98
19907.98 19175.98
17994.80
20454.08
18874.19
20042.05
17679.36
19192.64 19262.74 19907.98
974.92
5.08
PICKUP 1
108.81
111.37
111.37
111.71
111.71
111.37
111.71
Individual
nozzleperformance
performanceofofthe
theprinthead
printhead
Individual
nozzle
Individual nozzle performance of the
printhead
PICKUP
PICK
PICKUP
1 1 PICK
UPUP
2 2
10 10
Mean
Mean
%CV
%CV
Mean
21693.65 19704.73 21184.50 20773.02
Inter
batch
Analysis
- 2- days
Inter
batch
Analysis
2 days
20509.18
21184.50
20509.18 21693.65
21693.65
19704.73
21184.50
Inter19704.73
batch
Analysis
- 2 days
20451.71
20451.71
19469.52
19469.52 21150.03
21150.03 19249.51
19249.51 20652.58
20652.58
20509.18
21693.65
19704.73
21184.50
20451.71
Inter
batch
– –1 day
Inter
batchAnalysis
Analysis
1 day
19469.52
21150.03
19249.51
20652.58
17994.80
20042.05
17994.80 20454.08
20454.08
18874.19
20042.05
Inter18874.19
batch
Analysis
– 1 day 19175.98
19175.98
20509.18
Batch 1
Mean
Mean
Standard
Standard
Mean
deviation
deviation
Standard
(Sdev)
(Sdev)
deviation
PICK UP 2
PICK
PICK
UPUP
33
AVERAGE
AVERAGE
PICK UP 3
AVERAGE
Coefficient
Coefficient
of variability
of variability
- CV%- CV%
Coefficient
ofintensity)
variability
- CV%
(Sdev/Mean
(Sdev/Mean
intensity)
* 100%]
* 100%]
(Sdev/Mean intensity) * 100%]
9 9
10
8
8
9
7
7
8
6
6
7
5
5
6
4
4
5
3
3
4
2
2
3
1
1
2
0
0
1
102107
107112
112117
117122
122127
127
2 2 7 712 1217 1722 2227 2732 3237 3742 4247 4752 5257 5762 6267 6772 72 7777 8282 8787 9292 9797102
0
Nozzles
2
7 12 17 22 27 32 37 42 47Nozzles
52 57of of
62 the
67printhead
72 77 82 87 92 97 102 107 112 117 122 127
the
printhead
Nozzles of the printhead
Figure
3: Individual
nozzle
performance
spots
printed.
Figure
3: Individual
nozzle
performance
forfor
spots
printed.
individual
nozzle
performance
was
analysed
calculating
mean
coefficient
variability
(%CV)forforthese
thesepositions.
positions.The
TheCV
CV
TheThe
individual
nozzle
performance
was
analysed
byby
calculating
thethe
mean
coefficient
ofof
variability
(%CV)
Figure
3:
Individual
nozzle
performance
for
spots
printed.
values
were
calculated
to
be
4%
or
lower
indicating
highly
precise
and
consistent
individual
nozzle
performance.
values were calculated to be 4% or lower indicating highly precise and consistent individual nozzle performance.
The individual nozzle performance was analysed by calculating the mean coefficient of variability (%CV) for these positions. The CV
values were calculated to be 4% or lower indicating highly precise and consistent individual nozzle performance.
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Oremail:
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For more
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go
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Or
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information
go to:
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or email: [email protected]
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