Nano-PlotterTMMicroarraying and Picolitre Pipetting
MICROARRAYING
MICROARRAYING
Microarraying and Picolitre Pipetting
Non-contact spotting of picolitre droplets ensures utmost
flexibility and quality for the fabrication of complex
biochips. For many years, the GeSiM Nano-Plotter has been
a reliable companion for both research and production.
The GeSiM Nano-PlotterTM is not just another
microarray spotter. Modular hardware and
an open software guarantee high flexibility
to tackle even the most demanding print job.
GeSiM’s own piezo dispensers handle a wide
range of liquids, not only protein and DNA,
and eject droplets as small as 50 picolitres.
Microscope-based image processing makes
printing onto microelectrodes or other tiny
elements easy. Customised Nano-PlotterTM
versions, e.g. for integration into automated
production lines, are available on request.
A few applications
� Microarrays for research, diagnostics and
drug discovery (DNA, proteins, carbohy-
drates, lipids etc.)
� Spotting of beads and living cells
� Spotting into 96-well plates and onto
many other substrates
� Pipetting of miniaturized assays
� Coating of biosensors, (based on e.g. mi-
croelectrodes), aided by automatic image
processing
Key Features
| “Drop on demand” ensures arbitrary spot layout (including
unlimited replicates)
| No scratches on sensitive surfaces (slides, membranes,
microtitre plates, sensor chips…) via non-contact technology
| Homogeneous spots
| Spot volumes can be changed by varying the number of
drops per spot
| No need to adjust samples to pipettes: either vary pipetting
parameters or choose from different dispenser types
| “Spotting on the fly” for ultimate speed
| Dew point pipetting by chilling and humidification
| Alternative dispensers available for large volumes or viscous
media
|Countless accessories, e.g. automatic target recognition and
post-spotting array repair via microscope camera
www.gesim.de
GESIM Nano-PlotterTM
Configuration
The Nano-PlotterTM comes in two different
sizes with the following common features:
� Small footprint
� Dust cover included
� Z-sensor for height profile measurement
of targets
� 1 — 16 independent piezoelectric pipetting
tips with full fluidic control
� Different tip layouts on the pipetting head
(pitch in parentheses):
1 x 8 (4.5 mm), 1 x 6 (9 mm),
2 x 8 (9 and 4.5 mm), custom tools, etc.
NP2.1: Small platform
for max. 55 slides
NP2.1/E: Large platform
for max. 120 slides
Technical Data Nano-PlotterTM NP2.1 Nano-PlotterTM NP2.1/E
Dispensing area / slide tray
(width x depth, outer dimensions)
302 mm x 400 mm 645 mm x 400 mm
Traversing range in XY-direction (width x depth) 427 mm x 341 mm 777 mm x 341 mm
Footprint main unit 623 mm x 509 mm 973 mm x 509 mm
Height (incl. cover) 375 mm 375 mm
Weight (incl. cover) 30 kg 50 kg
Max. pipetting height 50 mm
Pipetting channels 1 ... 16
Power supply 110...240 V, max. 200 W
� Sample aspiration from 96-well or 384-
well microtitre plates (MTPs)
� XY repetition accuracy much better than
± 10 µm, encoder-controlled, step width
2 µm
� Travel speed up to 500 mm/s
� Function test in stroboscope of each
pipette tip before/after spotting
� Removable target tray
� Array density up to 3000/cm2
� Extra fast travelling mode and spotting on
the fly (spotting while travelling)
Work decks, cooling and humidification
The area inside the hood is variable. Different
removable slide trays (also tailor-made) and
trays for MTPs are available so that one can
trade in slide capacity for source MTPs.
NP2.1/E: Configurations for 5 x 23 slides with one
sample plate (top) and 3 x 23 slides with four sample
plates (bottom). More layouts are available.
Slide trays have handles and lock into place.
An additional tray saves time, as you can load
and unload one tray while the other one is in
the machine. A chiller plate with temperature
sensor allows the cooling of slides, e.g. to
keep spots moist.
NP2.1: Standard layout for 5 x 11 slides (up to 26
mm x 76 mm) and one sample plate
Slide tray removed from cooling chuck
System Platforms
MICROARRAYING
SUBSTANCES SUCCESSFUL-
LY DISPENSED BY GESIM’S
PIEZOELECTRIC PIPETTING
TECHNOLOGY:
Acetic anhydride, acetone
(propanone), acetonitrile
(ethanenitrile), betaine
(N,N,N-trimethylglycine), up to
3 M (up to 1.5 M for Pico-Tip),
chloroform (trichloromethane),
cyclohexanone, dextran solutions,
detergents (e.g. 0.02 % Genapol
C050, 2 % Triton X-100, 2% Tween-20),
dichloroben- zene, dichloromethane + 2 %
trichloroacetic acid, N,N-dimethylformamide (DMF), all mix-
ing ratios with water, dimethyl sulfoxide (DMSO), 1,4,-dioxane, DNA (PCR product < 2 kbp < 3 mg/ml in
buffer, plasmid < 4 kbp < 1 mg/ml), ethanol, ethylene glycol, moderate concentrations, N-FMOC-Ile-OH,
200 mM in DMF, up to 40 % glycerol in water, isopropanol (2-propanol), iodine in tetrahydrofuran/
pyridine/H2O 3:75:20:75 (v/v), liquid crystals (Merck Licristal® ZLI-2222-100 and Merck Licristal® MLC-
6681), MALDI matrix (alpha-cyano-4-hydroxy-cinnamic acid in NMP or 40 mg/ml 3-hydroxy-picolinic
acid / 6 mg/ml diammonium citrate in 20 % acetonitrile), methanol, 1-methylimidazole (N-methylimi-
Decks with vacuum or spring fixation exist,
but are only useful for pin spotting, as the
smooth touch-less operation does not move
slides. Decks holding microtitre plates (6 for
NP2.1, 12 for NP2.1/E) are available to pro-
duce microarrays in microtitre wells.
The standard slide tray has locating pins for
accurate slide positioning. Turning the bars
upside down creates a flat surface onto which
any substrate can be placed. And the space
between the bars can be magnetic (option),
helping to fix membranes by metal strips.
Of course, cooling of source plates and humid-
ification is possible to reduce evaporation and
to allow pipetting at the dew point..
Wash cycle in the normal (top) and the optional extra
wash station (bottom)
GeSiM’s piezoelectric tips are manufactured
from silicon and glass by micromachining
(reactive ion etching, anodic bonding etc.).
Samples come only in contact with silicon
dioxide (the Si surface is oxidized).
Each tip is individually actuated; sample
uptake and washing are done by syringe
pumps. The minimum dead volume (additional
sample) is approx. 1 µl. Unused sample can be
returned to the well after spotting. Various ar-
rangements on the pipette head are possible.
Tips are thoroughly rinsed on the inside and
the outside to avoid contamination. DNA solu-
tions (e.g. 0.1 mg/ml oligos) are washed away
in seconds. Proteins and peptides may need
longer, and in special cases additional wash
fluid (detergent, acid, base etc.) is taken from
a reservoir, a well or a second wash bowl.
Solids dissolved in organic solvents can be
spotted by putting a solvent “plug” and an air
gap between sample and system water.
Piezo tips for different droplet volumes
Nano-Tip J 0.35…0.6 nl
Nano-Tip A-J 0.2…0.4 nl
Pico-Tip J 0.05…0.08 nl
Special pipette tips e.g. Nano-Tip HV-J (for
highly viscous samples)
Nano-Tip H-J (heatable)
Nano-Tip AR-J (thinned for
printing into 96-well MTPs)
Two-row pipette head for up to 16 piezo dispensers
Piezoelectric Pipetting
GeSiM Piezo Pipettes
Slide deck, coolable micro-plate holders and
smoothly operating ultrasonic humidifier
www.gesim.de
GESIM Nano-PlotterTM
Stroboscope check of the piezo tips
Each piezoelectric tip is tested by analysing
the stroboscope image of the flying droplets
before and after spotting. If no droplet is
detected, maybe caused by an empty well,
the pipette is switched off and the array is
repaired later. (If dry spots are visible, a
microscope can search for missing spots!)
Droplet volume and speed depend on the
sample (mainly viscosity). Computing a body
of rotation from the droplet contour gives a
rough volume estimation; the speed is meas-
ured by analysing the droplet pattern at two
time points. You can thus optimize pipette
parameters so that all samples are spotted
with the same speed or the same volume.
If GesiM’s microfluidic flow sensor is placed in
the tube system, it can measure the flow dur-
ing the stroboscope test. This provides very
accurate droplet volume determination.
VIDEO
Probenaufnahme Dosierparameter Spotting Entleeren/Waschen Waschen mit Sample
aspiration
Optimising dispense
parameters and
functionality test
Additional rinse cycle
with extra wash fluid
(optional)
Spotting Emptying and
washing
Stroboscope image (left), 8-fold flow sensor on a
syringe pump unit (centre) and flow sensor graph
(right). A flow rate of 1.7 µl/min at a frequency of
100 Hz corresponds to a droplet volume of 280 pl.
Viscous samples and large volumes
GeSiM manufactures a variety of other dis-
pense heads. Viscous media can be delivered
by our heatable piezo dispensers. We also of-
fer heatable systems for bulk piezo pipetting
with large sample reservoirs. And finally there
is a solenoid valve dispenser for nanolitre
volumes.
Third-party dispensers with piezo or pneu-
matic valves can also be used; they work at
viscosities of up to a few thousand mPa⋅s.
And yes, even pins.
The Nano-PlotterTM syringe unit can also be
used for bulk displacement dispensing. Pas-
sive capillary tips or adapters for disposable
tips are available on request. Please consult
our Nano-Plotter catalogue for details and
more spare parts..
Special dispensing systems from GeSiM. Left, heatable piezo tip that is compatible with microtitre plates, with
additional temperature sensor cable in a Nano-Plotter. Centre, heatable bulk piezo dispenser with reservoir
(without fluidics). It can deliver droplets as small as 10 pl. Right, glass capillary dispenser with solenoid valve for
higher viscosity and volumes of > 50 nl. Needs a fluidic system with slight overpressure.
Pipetting Cycle
MICROARRAYING
Special Equipment and Software
Software and programming
The intuitive Nano-PlotterTM control software,
NPC16, supports all configurations, but you
don’t have to care for this, as the system
comes pre-configured.
NPC16 features a scripting tool (Nano-Plot-
ter Language) with arithmetic expressions,
constructs like loops and branches, and full
hardware access; so you can develop your
own “NPL programmes”. This is an option, but
rarely needed, as all necessary programmes
are included, ready for use.
Standard procedures
Service routines exist for overnight standby
with occasional washing to keep pipettes
happy, a cleaning procedure for dirty pipettes,
or semi-automatic adjustment of pipettes to
the same droplet speed, to name a few.
Our standard spotting routines should address
all your needs no matter how pipette head or
targets are set up, require zero programming
and feature extensive data logging for quality
control, including saving of stroboscope
pictures and determining droplet volumes and
speeds before and after spotting.
Transfer list: Programmes are not restricted to
a certain order (as in the left two panels). In
fact, sample wells can be assigned to arbitrary
spot positions in one or more sub-arrays, also
with replicates. This is described in a “transfer
list” text file, but you can use a graphical user
interface by clicking on source and target
positions. But given the many options, typing
is often easier than clicking.
Sequential spotting: ”Drop on demand” allows
to spot only one sample at a time. Although
slow, this flexible mode allows any array
layout and adjustment of inter-tip positions.
Spotting with automatic target finding via a
microscope must always be done sequentially.
Simultaneous spotting: This is the fastest
method, as all tips fire at the same time. Spots
must be arranged in the pitch of the pipette
tips (9 or 4.5 mm), so spots in between are
printed in subsequent cycles. Array positions
are again given by a (different) transfer list.
Thanks to our customers, our “TransferSim”
programme has matured to a “Swiss Army
Knife” with countless options, e.g. elaborate
wash procedures, sample uptake using only
a subset of pipettes, spotting only on certain
slides or sub-arrays, solvent delivery, ”spot-
ting on the fly”, varying pipetting parameters
and much more. Multiple source plates, with
or without manual exchange, are supported.
Parallel spotting is also the only choice when
spotting into MTP wells.
Spotting onto tiny structures
It is not easy to dispense a 60 µm droplet
exactly onto a target of similar size. A micro-
scope camera on the print head helps: first
spot positions on all targets are identified
(typically via alignment marks), then the X/Y
position of the droplet is measured via a test
spot to correct even the smallest deflection.
The pattern recognition works well for pure
two-dimensional targets such as microe-
lectrode pads or straight conducting paths,
but also for nano-wells, microcantilevers,
microfluidic channels and more. Missing spot
detection and repair is also possible; live spot
detection during printing on a light table will
soon be available.
This software extension allows the unlimited definition
of spot positions relative to alignment marks. On
the right: microscope above a chip and example of a
successfully detected microelectrode array.
Handling of thousands of samples
All Nano-Plotters can be combined with
plate hotels. This here from Thermo allows
automatic handling of up to 56 MTPs. Plate
storage is temperature- and humidity-con-
trolled.
A lid handler guar-
antees the transfer
of covered MTPs,
especially for quickly
evaporating samples.
www.gesim.de
GESIM Nano-PlotterTM
Examples from Research and Manufacturing
0,1 mg/ml Oligo
10 drops = 3 nl
100 drops = 30 nl
2 m
m
Arrays for DNA, proteins and more
GeSiM’s piezo technology provides high spot
homogeneity and allows different spot sizes
simply by varying the number of drops per
spot. Different samples can be spotted by ad-
justing pipetting parameters (mainly voltage).
For higher viscosity, “high-viscosity” tips
(“HV-J”) and heatable tips are available.
The spotting pitch depends on the surface.
Hydrophobic substrates allow grid sizes down
to 150 µm. Piezoelectric tips are perfect to
process typical buffers such as 3x SSC, phos-
phate-buffered saline (PBS) or Tris, but also
up to 40% glycerol.
The Z-height sensor measures all target Z-positions
for automatic adjustment of spotting heights
Spotting onto microstructured substrates is
possible. Using piezoelectric, solenoid valve
or displacement (“passive”) dispensing, nan-
olitre and microlitre volumes can be spotted
onto e.g. conducting MALDI targets. Automatic
centring is possible, if image processing is
used (see above).
Microarrays can be generated in three-dimen-
sional objects, e.g. MTP well bottoms.
Spots on a membrane (2.5 nl, 0.4 mm pitch) and in a
96-well MTP at 0.3 mm pitch
Large-scale chip production
Of course, the rather small Nano-PlotterTM
is ideal for research and chip development,
but this does not mean that it is incapable of
high-volume chip production.
The picture on the right shows a former
production site for specially designed FDA-
cleared diagnostic chips of a company in Utah.
And a growing number of companies in China
rely on the Nano-PlotterTM for bulk biochip
production.
Sensors
Numerous labs all over the world use the
Nano-PlotterTM for special operations such as
the production of (bio)sensors, where (bio)
molecules are spotted onto small electrodes,
optical structures, microcantilevers or similar.
This requires very precise positioning that is
aided by a microscope and automatic image
processing (here an example from Scotland).
The microscope can also be used in combina-
tion with other dispensing systems such as
pin tools or a dispenser for adhesives.
Gesellschaft
für Silizium-Mikrosysteme mbH
Bautzner Landstraße 45
01454 Radeberg, Germany
Tel. +49–351–2695 322
Fax +49–351–2695 320
For more information (applications,
systems, distributors etc.) please visit
www.gesim.de
Specifications subject to
change without notice
20
19
GESIM Nano-PlotterTM
We are not licensed under any patents owned
by Oxford Gene Technology Limited (OGT) or
related companies and cannot pass any such
licence to our customers. A licence under
OGT’s patents may be necessary to manufac-
ture or use oligonucleotides arrays.
GeSiM mbH