JUST PUSH PRINT

Biodevice Printing Using Bioinks, Electroinks and Quantum Dot Inks

Jan Lawrence Sumerel and Kai Sudau

FUJIFILM Dimatix, Inc.,2230 Martin Avenue, Santa Clara, California 95050-2704, U.S.A.

Keywords: Ink jet, piezoelectric, biomaterials, microarray, bioink, electroink, quantum dot.

Abstract: Many advanced medical and environmental test devices require microscale patterning of cells, proteins, or

other biological materials, and the need for these devices to contain active functional material components

has increased dramatically. In addition, the biological material oftentimes requires an interface with an

electrical or optical output signal. Efficient production methods are paramount to meeting market demands,

and ink jet printing offers an easy, low cost alternative to materials deposition used in current biodevice

manufacturing. However, fluid development and proper printing parameters at the research level are

required for manufacturing processing and will be critical to process adoption. In this paper, operating

parameters and fluid characterization have been developed through processing biomaterials, organic and

inorganic conductive fluids and semiconductor nanoparticles. Because of the inherent versatility,

uniformity and scalability of this system, established operating parameters coupled with proper fluid

characterization will ultimately be translatable to production line systems of biodevice components.

1 INTRODUCTION

Biological monitoring devices and medical devices

generally have two material components used in a

stepwise fashion, a biological material that works as

both a reaction beacon and the biochemical reactive

site followed by an optical, piezoelectric or

electronic material that amplifies the signal to allow

a measurable readout of the reaction. Typically,

manufacturing protocols are distinct for each

component, but ink jet printing can be used for the

deposition of both materials. Interfacing processing

of these two components may be critical to

biodevice success, and electronic file pattern

formation allows component alignment. Ink jet

printing is inherently compatible to high throughput

(Antoniadis, 2007). Already, interesting

technological phenomena have spawned from the

patterning of structurally and functionally different

materials including high performance ceramics

Lewis, et al, 2006). For this reason, drop-on-

demand ink jet printing, a simple fabrication

process, has become a prominent player in materials

processing for biodevice components (Padinger,

2007). However, it is a big step for biodevice

developers to jump into robust in-line manufacturing

production systems. This type of equipment

requires a sizable financial investment plus

sufficient experience so that manufacturing

specifications and in-house knowledge can be

established. Thus, a low cost, easy-to-use laboratory

scale system is required for preliminary

experimentation. This strategy then allows substrate

evaluation, on-site development, and fluid

manufacturing to all occur simultaneously. This

need has been addressed with an R&D tool that

offers printhead maintenance, substrate alignment,

nozzle inspection and drop analysis, and ease-of-use

(Sumerel et al, 2006).

Just push print, the most common command for

the desktop printer, can now be used in the

laboratory or in manufacturing lines. Ink jet printing

is a simple and cost effective technique with

applications in the fields of electronics and

biomedicine and has been shown to have specific

applications in these industries (Sirringhaus et al.,

2000, Calvert, 2001, Haber et al., 2005). In contrast

to other multi-step production methods, ink jet

printing is an additive process that precisely deposits

metered quantities of fluid onto a variety of

substrates including glass, silicon, plastics, organic

thinfilms, and metals based on a user generated

pattern. The resolution of the printed pattern is

determined by a number of factors, including

297

Lawrence Sumerel J. and Sudau K. (2008).

JUST PUSH PRINT - Biodevice Printing Using Bioinks, Electroinks and Quantum Dot Inks.

In Proceedings of the First International Conference on Biomedical Electronics and Devices, pages 297-303

DOI: 10.5220/0001054902970303

 SciTePress

substrate/fluid contact angle, nozzle size, and lateral

resolution of the printhead (Song and Nur, 2004).

Ink jet printers can dispense fluid drops with

volumes in the picoliter (ρL) to microliter (µL)

range, and an integral step in bringing this

processing technique from the laboratory to

manufacturing systems is the development of

jettable fluids. The chemical properties of the fluid,

including density, surface tension and viscosity,

determine its jettability (Sumerel, et al., 2006).

During drop formation, energy is distributed

between the fluid’s viscous flow, surface tension,

and kinetic energy (Xu et al., 2005). The deposited

fluid volume is directly proportional to nozzle size.

This flexibility enables microscopic patterned

thinfilms of functional materials at a variety of

resolutions. The physical properties of the patterned

thinfilms (film thickness and pixel values) are

dependent on the fluids coupled with the drive

electronics of the printing device. In general, 2D

drawings, pictures or structures, formatted as a

bitmap image, can be translated into X and Y print

coordinates for materials deposition (drop-on-

demand). Each individual nozzle ejects a drop with

a ligament. The ligament and the drop coalesce

during flight to make a volumetric sphere and upon

contact with the substrate, the sphere alters its three

dimensional structure to become columnar. The

resulting printed image is a compilation of drops

where the third dimension is equal to film thickness,

a physical property that is dependent on particle

loading, drop spacing and drop spread. Once this

critical but iterative R&D phase of process and

material evaluation is complete to allow sustainable

ink jet printing, the fluids are scalable for production

use.

1.1 Ink Jet Printing Employing MEMS

Devices

The required heating process for thermal ink jet

printing (300°C) will damage thermally-sensitive

materials, thereby limiting their use in devising

functional devices (Calvert, 2001, Xu et al., 2005).

In contrast, using piezoelectric ink jet printing,

thermally sensitive materials are deposited under

ambient conditions. Piezoelectric printheads contain

a lead zirconate titanate (PZT) piezoelectric ceramic,

nozzles, and a fluid chamber. When a voltage is

applied to the PZT, mechanical vibrations create

acoustic waves that in turn force fluid out of the

chamber through the nozzles (Brünahl and Grish,

2002). Piezoelectric printheads are categorized

based on the deformation mode of the PZT (e.g.,

squeeze mode, bend mode, push mode, or shear

mode) (Myatt et al., 2006). MEMS fabrication has

increased the precision and resolution of the

deposited materials (Menzel, C., 2005). These

silicon devices increase jet-to-jet uniformity and

drop placement accuracy. The inertness of the

silicon expands the operating ranges to allow higher

chemical diversity and fluid throughput expanding

piezoelectric ink jet printing from the ability to print

graphic inks to the realm of printing functional

fluids required for biodevice manufacturing.

The ink jet printhead is powered by a

piezoelectric unimorph, which is constructed in the

plane of the wafer and consists of patterned PZT

bonded to a silicon diaphragm (Brünahl and Grish,

2002). The effective diameter of the nozzle is 21.5

mm; this nozzle size is approximated to generate 10

ρL drops. An important operating parameter of this

particular device is the negligible void volume due

to the direct fluid/printhead interface.

Fluid flow properties like low viscosities, low

boiling points, high surface tensions and non-

Newtonian behaviors are hallmarks of functional

materials and are also generally unfavorable

chemical characteristics for ink jet printing.

Manipulating the parameters that generate the

electronic signal to drive the movement of the PZT,

including its frequency, wave shape, wave duration

and voltage has provided a significant advancement

in printing an array of functional materials and has

been one of the areas of our research. The ability to

adjust the jetting parameters has been critical to the

success of printing bioinks and electroinks.

1.2 Functional Fluid Deposition

Thermal ink jet printing has been employed for the

deposition of biomaterials (Xu et al, 2005, Setti et

al., 2005). A glucose biosensor was fabricated by

thermal ink jet printing, and the enzyme, glucose

oxidase was made into a biological ink using

phosphate buffer and 10% glycerol (Setti et al.,

2005). In contrast to piezoelectric ink jet printing

where there are requirements for viscosity (8-14

centipoise (cps)) and surface tension (28-32

dynescm

-1

), most biological materials exhibit very

low viscosities (1 cps) and very high surface tension

values (60 dynescm

-1

). In addition, biological fluids

generate steam at high temperatures just like inks in

thermal ink jet printers. This heating process causes

bubble formation and fluid output at the nozzle plate

(Bae, et a., 2005). Major fabrication advances have

been made using thermal ink jet printing (Lemmo et

al., 1998) due to the low cost and wide availability,

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and at first glance this method should simplify

biomaterials deposition due to the looser

requirements for fluid formulation. However, the

thermal ink jet process may cause damage to

thermally-sensitive materials used in biology and

medicine. In contrast, piezoelectric ink jet printing is

a thermally constant process and does not require

heat thereby increasing the chances of biomaterial

stability.

Rapid detection employing microarray methods

are necessary to biomedical and chemical sciences

(Diehl, F., 2002). Miniaturization and automation of

arrays may lead to decreased costs and faster

analysis times (Peck, 2007). As drop sizes decrease,

feature sizes decrease and array densities increase.

Many forensic samples obtained in the field have

restricted amounts of recoverable material, and in

some cases, two polymerase chain reactions are

required to reach the levels of sensitivity and

verification required in the amplified

deoxyribonucleic acid (DNA) product (Vuorio et al.,

1990). Since piezoelectric ink jet printing only

requires 10 ρL per sample, the amount of DNA

needed for a precise polymerase chain reaction assay

is greatly reduced. For example, this technique may

provide an important advance in studying the

variation in a segment of mitochondrial DNA (a

non-coding region between two transfer ribonucleic

acid genes) (Salas et al., 2001). Variations in this

section of the gene is one obvious choice for

forensic identification because of sample number

(Wilson et al., 1995), and the predominant isolated

sample, human hair, shows low keratin protein

variation between individuals (Rodriguez-Calvo et

al., 1992). Unlike contact printing techniques (e.g.,

pin spotters) or expensive industrial ink jet

printheads, the single-use ink jet printhead

technology requires minimal deposition of fluids and

minimal cross-contamination (Manning, H., 2007).

The need for continual reiterations of circuit

design gestures for new approaches away from the

reductive process of masking and etching to create

metal patterns (Hwang, 2002) towards additive

processing. There are also market drivers for organic

electronic materials due to their adherence to

substrates, flexibility, performance and the ability to

process these materials at low temperatures (Shaw

and Seidler, 2001). Ink jet printing provides the

necessary technological platform to increase

throughput and lower processing costs. Indeed, low

capital costs, process simplicity, and flexibility have

been the important attributes that make this

technique practical for conductive trace patterning.

Many conductive precursor fluids are being jetted

using both thermal and piezoelectric ink jet printers

(Sirringhaus et al., 2000, Sawhney et al., 2006, Teng

and Vest, 1988; Volkman, et al., 2004). In fact, ink

jet printing is considered one of the key technologies

in defined polymer deposition (Sawhney, 2006).

Polymer structural confirmation varies with

temperature thus ambient processing conditions are

required. At low temperatures, typically below the

glass transition temperature (T

), polymers maintain

their natural, globular structure. At higher

temperatures, above the T

, they swell into open

conformations, essentially breaking their

entropically favorable π-π interactions (Baiesi et al.,

2001). With the conformational collapse, the

material becomes less conductive. In order to move

towards feasible ink jet manufacturing processes for

either conductive polymers or metallo-organic

fluids, initiating formulation, printing and post-

processing techniques are required. The fluids must

maintain solvent monodispersity; once printed, they

must properly adhere to the surface (Mei et al.,

2005). These criteria are integral for successful

printing, for even the smallest amount of

discontinuity will make the material non-conductive

and lower its mechanical strength.

Early attempts at ink jet printing silver metallo-

organic fluids capitalized on its advantageous

annealing temperature post-printing (200°C). The

resultant silver conductive traces on a variety of

materials including flexible substrates and substrates

are left with a low thermal budget (Volkman et al.,

2004). The direct writing of silver ink onto a grid

pattern of solar cells has been previously done using

a self-built printer and a Siemens ink jet printhead

(Teng and Vest, 1988). They modified their

printhead by machining restrictive nozzle plates that

varied drop size. The printer was run between 100

and 200 Hz which resulted in a printing speed of a

few cmsec-1. This single laboratory technique was a

slow throughput process and required multiple

printing cycles for effective deposition, so although

it is not agreeable to manufacturing protocols, it was

an important proof of concept step.

The choice of additional organic material in the

starting fluid greatly influences the obtained

conductivity (Mei et al., 2005). Once printed, the

silver in the fluid must be annealed to convert the

nanoparticles to a bulk silver thinfilm so that the

resistance values can closely mimic bulk silver. The

resulting amount of silver per volume of fluid is

controlled by the annealing temperature cycle

controls, and the effect of the organic decomposition

into the gas phase during annealing determines the

porosity of the printed material, which affects its

JUST PUSH PRINT - Biodevice Printing Using Bioinks, Electroinks and Quantum Dot Inks

299

continuity (Mei et al., 2005). Additionally, the

proper jetting parameters required for high

performance printing is fundamental for

reproducible deposition. The final feature size of the

material on the substrate is determined by these

parameters, and the overall conductivity is

established according to the applied thermal

processing (Mei et al., 2005).

2 MATERIALS AND METHODS

Proteins were dissolved in phosphate buffer saline

solution (Fisher Scientific, Fair Lawn, NJ, USA) and

1.6 µM solution. 1 % of polysorbate 20 surfactant

(Fisher Scientific, Fair Lawn, NJ, USA). 10 mg/mL

of human genomic DNA was dissolved in 50%

ethanediol.

Two percent (2%) glycerol (Sigma Aldrich, St.

Louis, MO) was added to a poly (3,4-

ethylenedioxythiophene) poly(styrenesulfonate)

(PEDOT/PSS) aqueous dispersion (H.C. Stark,

Goslar, Germany). ANP Silverjet nanopaste

(Advanced Nanoproducts, Chungcheonguk-do,

Korea) and Cabot Inkjet Silver Conductor (Ag-Ij-G-

100-S1, Albuquerque, NM) were used as packaged.

Fluids were sonicated in a water bath in a Branson

1510 sonicator at room temperature using highest

sonic level for 30 minutes. Fluids were degassed for

2 hours at 5 mbar pressure in a degassing chamber.

Quantum dots were obtained from UT Dots

(Savoy, IL, USA) and were serially diluted in 53%

polypropylene glycol 400, 45% propylene carbonate

solution containing 0.01% tetramethyl-5-decyne-4,6-

diol, 2,4,7,9-propanol (Surfynol 104PA; Air

Products, Allentown, PA).

Clean glass wafers were purchased from VWR

(VWR Scientific, West Chester, PA). Both Kapton

(Dupont, Wilmington, DE) and Teslin

synthetic

thinfilms (PPG Industries, Pittsburgh, PA) were kept

clean after purchasing and cut into 8 x 11 inch sheets

using laboratory scissors that had been cleaned with

70% ethanol (Sigma-Aldrich, St. Louis, MO).

Single-side polished 150 mm silicon 100 wafers

were obtained from Silicon Quest International

(Santa Clara, CA) and sputtered with 300 nm gold

layer using an Au target and a converted TES

sputterer.

The DMP-2831 (FUJIFILM Dimatix, Santa

Clara, CA) was used according to packaging

instructions. Contact angle measurements were

carried out using a VCA Optima XE (AST,

Billerica, MA). 2 µL samples were manually

pipetted for the measurements.

Scanning electron micrographs were obtained

using a Philips XL30 ESEM. Resolution was

obtained based on operating voltage of 5 kV.

Tapping mode AFM was conducted on a Digital

Instruments Dimension 3100 using an etched silicon

tip with a nominal radius of curvature of 10 - 20 nm.

Scan sizes were varied, depending on the feature

size. The scan rate was 0.1 - 0.3 Hz. The set point

was set to 60 - 70% of the free-standing root mean

square of the voltage of the oscillating tip.

Resistance measurements were obtained using a

Fluke 110 True RM multimeter. Anode was put at

one end of silver contact on glass wafer and cathode

was placed on top of other end of silver contact.

Electrodes were manually held during

measurements.

3 DISCUSSION

Both DNA and proteins were ink jet printed after

fluid formulation trials. Human genomic

deoxyribonucleic acid was printed in 10 ρL of 50%

ethanediol in the bottom of a 384 well assay plate or

onto a silicon wafer in a 254 µm grid with high

fidelity drop formation and uniform drop speed

(Figure 1). The high surface tension of the DNA in

water was mediated by the lower surface tension of

the 1,2 ethanediol (47 dynescm-1) but the short

ligaments in Figure 1 demonstrated how the surface

tension of the fluid is still the predominant force in

drop formation.

Figure 1: Deoxyribonucleic acid leaving the nozzle plate.

The only way to successfully print this fluid was by

lowering the jetting frequency and extending the

wave pulse from 11.52 µs to 14.1 µs. After

successful printing, the samples were air-dried and

tested for polymerase chain reaction amplification.

A 750 base pair fragment was resolved on a 1.2%

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agarose gel and visualized using ethidium bromide

(data not shown). These successful results suggest

that a significant cost savings may be obtained by

using piezoelectric ink jet printing for the detection

of clinically or environmentally relevant DNA

species.

In general, globular proteins are stable in

phosphate buffered saline with a small amount of

non-denaturing detergent. We employed 1 % of

polysorbate 20 surfactant, a detergent that is often

used in protein purification due to its biochemical

compatibility and protein stabilization (Sumerel et

al., 2001). It has a second chemical attribute that it

lowers the surface tension of the fluid. Bovine

serum albumin was printed in 10 ρL drops, and then

an α-BSA polyclonal antibody (Sigma Aldrich, St.

Louis, MO) labelled with Cy3 was printed in

random array over the protein drops. The protein

array is shown where the antibody/antigen reaction

is shown in green and protein alone is shown in blue

(Figure 2).

Figure 2: BSA protein incubated with a-BSA polyclonal

antibody labelled with Cy3.

Due to appropriate fluid formulations required for

ink jet printing, glycerol was added to the stock

PEDOT/PSS solution to increase fluid viscosity. A

waveform was employed for successful PEDOT/PSS

printing (maximum jetting frequency of 1.0 kHz for

a pulse width of 17.0 µs). This waveform is a

critical parameter for jetting this particular fluid.

The applied voltage was tuned specifically for

each nozzle to provide uniform jetting speed to

ensure reproducible drop volumes. Images of the

jetting fluid were captured by light micrographs

using this camera and software system (Figure 3).

In panel A, the fluid is leaving the nozzle with the

ligament still evident. In panel B, at 500 µm, the

ligament has drawn into the drop, and the fluid is

flying towards the substrate at 9.25 msec-1, ten

times faster than the homebuilt printer discussed

above (Teng and Vest, 1988). The resulting

PEDOT/PSS on a silicon wafer pattern is shown

(Figure 4). The PEDOT/PSS spreads on an

untreated glass wafer with a contact angle of 18°

(data not shown).

Figure 3: Light micrographs of fluid jetting from printhead

nozzle and time of flights. A. Fluid leaving nozzle. B.

Drop formation at 500 µm.

Figure 4: PEDOT/PSS printed on silicon wafer. A. Bright

field. B. Dark field + UV.

Reliable printing procedures for two commercially

available conductive silver precursors have been

examined. Both fluids have ideal fluid flow

properties for ink jet printing and have higher than

50% silver nanoparticle load. The viscosity and

surface tension values of the ANP Silverjet

nanopaste are 9 cps and 26.5 dynescm-1

respectively. This fluid jetted at a maximum

frequency of 5.0 kHz with a pulse width of 13.2 µs.

Because of its high particle load (54%) and

uniform particle size as demonstrated by

transmission electron microscopy, low-temperature

annealing produces a traceable conductivity in the

printed material. Scanning electron micrographs

were obtained of the annealed printed nanoparticles

to compare the films produced (Figure 5).

Figure 5: Electron micrographs of ANP silverjet nanopaste

on Teslin

. Before annealing. B. After annealing.

Panel A shows the ANP Silverjet nanopaste on

Teslin

before annealing (silver on left, Teslin

right). With single-pass printing, the fluid makes a

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301

uniform film on the Teslin

substrate in spite of the

material’s surface roughness (Panel A). Figure 5,

panel B shows the same film on the same substrate

after annealing for 1 hour at 200°C (annealed silver

on left, Teslin

on right). Not only do the edges

look slightly more uniform, but full coverage of the

film on the substrate with single pass printing

created a very thin silver full coverage film on the

Teslin

. Because accurate feature measurements are

difficult on flexible substrates, feature thickness was

measured on gold-coated polished silicon nitride

wafers (contact angle 41.8º, data not shown). We

measured feature sizes using atomic force

microscopy (AFM) with features printed at 20 mm

drop spacing. Figure 6 shows the overall scan area

of a single row of drops (Panel A).

Figure 6: Atomic force microscope images of single row

of ANP silver nanopaste drops. A. Overall scan area of a

single row. B. 3D rendering C. Feature measurements.

The three-dimensional rendered view in Panel B

shows the overall jetting uniformity. Panel C shows

the calculated feature measurements. The width of

the feature was 40.6 µm, and the film thickness of

1.59 µm demonstrates the utility of producing

patterned thinfilms using ink jet printing technology.

Because electrical performance is often described in

terms of the bulk resistivity, resistance values were

measured after annealing the silver nanoparticles.

The resistance of the ANP Silverjet nanopaste is

1.1Ω, and the resistance of the Cabot Inkjet Silver

Conductor is 0.3Ω (data not shown). The low

resistance measurements in both cases were taken on

equally-sized patterns on identical glass wafers.

These values are in the same range as resistance

values obtained by Sawhney and colleagues.

In order to achieve even finer features in

manufacturing electronic applications, drop volumes

below 10 ρL are required. Employing 1 ρL

printheads, the reduced drop volume will produce 20

µm silver fluid features on Kapton

employing the

ANP Silverjet nanopaste fluid. The miniaturization

of drop volumes is demonstrated by ink jet printing

with the 10 ρL cartridge followed by ink jet printing

with a 1 ρL cartridge on the same wafer substrate

and performing scanning electron microscopy (data

not shown) where the radius of the 1 ρL feature is

more than twice as small as the radius of the 10 ρL

feature. Smaller drop volumes will lead to fine

conductive traces required for appropriate feature

sizes in photovoltaic and electronic applications and

directly address market demands.

Quantum dots were diluted in 53%

polypropylene glycol 400, 45% propylene carbonate

solution containing 0.01% tetramethyl-5-decyne-4,6-

diol, 2,4,7,9-propanol and printed onto a clean

silicon wafer (Figure 7) using an optimized

waveform.

Figure 7: Scanning atomic force image of quantum dots

printed on silicon wafer.

Although these particles are identical in synthesis

methods and solvent composition, their

concentration had a radical effect on their deposition

on a silicon wafer substrate. The 2.6 nm quantum

dots spread on the substrate whereas the 4.0 nm

quantum dots did not spread to the same extent. The

thinfilm produced by the 2.6 nm quantum dots was

only 50 nm thick whereas the thinfilm produced by

the 4.0 nm quantum dots was about 450 nm strongly

suggesting that contributions to the final 3D

structure of the thinfilm are particle-concentration

dependent.

4 CONCLUSIONS

The software interface and waveform tuning allowed

fluid process development for a scalable ink jet

printing process. Ink jet printing of biologically

active, electronic and semiconducting materials is a

cost-effective manufacturing process. The printing

parameters for these materials have been

demonstrated, and the resulting interaction with the

substrate demonstrates that ink jet printing is an

iterative process where the interplay between the

chemical properties of the fluid, the cartridge

assembly, the machine operating procedure, the

substrate and post-ink jet processing all determine

whether this process is viable. Printing of two silver

nanoparticle fluids has demonstrated the flexibility

of the Dimatix Materials Printer, and these silver

nanoparticles were successfully processed into

conductive traces. Fluid formulation and ink jet

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printing operating parameters are both key to the

success of ink jet printing functional materials.

Established operating parameters can now be

translatable to production line systems with built in

versatility, uniformity and scalability for biodevice

production. Future directions will be to incorporate

ink jet printing with circuit printing and biological

fluid deposition for single deposition biodevice

processing methods.

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