Nano-PlotterTMMicroarraying and Picolitre Pipetting

GESIM Nano-PlotterTM

Microarraying and Picolitre Pipetting

Non-contact spotting of picolitre droplets ensures utmost flexibility and quality for the fabrication of complex biochips. For many years, Nano-Plotters have been reliable companions for both research and production.

The GeSiM Nano-PlotterTM is not just another

microarray spotter. Modular hardware and an

open software guarantee utmost 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, usually 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

www.gesim.de

Configuration

The Nano-PlotterTM comes in two different

sizes with the following common features:

� Small footprint

� Includes dust cover

� 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 (9/4.5

mm), 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 ± 10 µm, encod-

er-controlled, step width 2 µm

� Travel speed up to 500 mm/s

� Function test in stroboscope of each pi-

pette tip after sample aspiration Remova-

ble target tray

� Array density up to 3000/cm2

� Extra fast travelling mode and spotting on

the fly (spotting while travelling)

Work decks, cooling and humidificationThe area inside the hood can be varied. Var-

ious 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.

Having more than one 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

GESIM Nano-PlotterTM

SUBSTANCES SUCCESSFULLY

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 (trichlorometh-

ane), cyclohexanone, dextran

solu- tions, detergents (e. g. 0.02 %

Genapol C050, 2 % Triton X-100, 2% Tween-20),

dichlorobenzene, dichloromethane + 2 % trichloroacetic acid,

N,N-dimethylformamide (DMF), all mixing 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

Decks with vacuum or spring fixation exist,

but are only useful for pin tool 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

produce 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 is magnetic, helping to fix

membranes by metal strips.

Of course, cooling of source MTPs and humid-

ification is possible to reduce evaporation and

allow dew point pipetting.

Wash cycle in the normal (top) and the optional extra

wash station (bottom)

GeSiM’s piezoelectric tips are made of 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 are spot-

ted 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

Stroboscope check of the piezo tipsEach 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.

If GesiM’s microfluidic flow sensor is placed

in the tube system, it can measure the flow

during the stroboscope test, This provides

a very accurate droplet volume determination.

VIDEO

Dilutorsystemfür Ver- und Entsorgung

der Systemüssigkeit

Probenaufnahme Dosierparameterprüfen und

Funktionstest

Spotting Entleeren/Waschen Waschen mit zusätzlicher Waschlösung

(optional)

Systemüssigkeit

Waschlösung

Probe

EIN PIPETTIERZYKLUS TEST / SPOTTEN WASCHENPROBENNAHME

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 volumesGeSiM 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 overpressure.

Pipetting Cycle

GESIM Nano-PlotterTM

Special Equipment and Software

Software and programmingThe 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-Plotter

Language) with arithmetic expressions, con-

structs like loops and branches, and full hard-

ware access; so you can develop your own

“NPL programmes”. But this is needed only in

rare cases, as all necessary programmes are

included, ready for use.

Standard proceduresService 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.

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 pic-

tures and determining droplet speeds before

and after spotting.

Transfer list: Programmes are not restricted to

a certain order (as in the left panel). Indeed,

sample wells can be assigned to arbitrary

spot positions in one or more sub-arrays,

with replicates. This is described in a “transfer

list” text file, but you can use a graphical user

interface and click 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 also be done sequentially.

Simultaneous spotting: This is the fastest

method, as all tips fire at the same time. Spots

are arranged in the pitch of the pipette tips (9

or 4.5 mm), so spots in between are spotted

in subsequent cycles. The array position is

again given by a (different) transfer list.

Thanks to the our customers, our “Transfer-

Sim” 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 de-

livery, “spotting 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 structuresIt is not easy to dispense a 50 µm drop-

let exactly onto a target of similar size. A

microscope camera on the print head helps:

first target positions are identified (typically

via alignment marks), then the X/Y position

of the droplet is measured to correct even the

smallest deviations.

The pattern recognition works well for pure

two-dimensional targets such as microelec-

trode pads or straight conducting paths, but

also for nano-wells, microcantilevers, micro-

fluidic channels and more.

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 samplesAll 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

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 moreGeSiM’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, special 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 productionOf 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.

Many companies in China also rely on the

Nano-PlotterTM for bulk biochip production.

SensorsNumerous 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,

microcantilevers or similar. This requires very

precise positioning that is aided by a micro-

scope 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.

GESIM Nano-PlotterTM

Gesellschaft

für Silizium-Mikrosysteme mbH

Bautzner Landstraße 45

01454 Radeberg, Germany

Tel. +49–351–2695 322

Fax +49–351–2695 320

info@gesim.de

For more information (applications,

systems, distributors etc.), please visit

www.gesim.de

Specifications subject to

change without notice

2018

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