Design Alternatives for Design Alternatives for Micropatterning of Micropatterning of MacromoleculesMacromolecules

GROUP 3:GROUP 3:Sailaja AkellaSailaja Akella

Caroline LaMannaCaroline LaMannaTeresa MakTeresa Mak

Rupinder SinghRupinder Singh

Advisor:Advisor:Emilia EntchevaEmilia Entcheva

Design ProjectDesign Project

Design a system that facilitates cell Design a system that facilitates cell deposition and micropatterning to be used deposition and micropatterning to be used in the creation of cellular and polymer in the creation of cellular and polymer based circuits.based circuits.

Customer CriteriaCustomer Criteria Suitable for printing proteins for cell Suitable for printing proteins for cell

adhesionadhesionFibronectinFibronectinCollagen IVCollagen IVCollagen I –fluorescence labeled Collagen I –fluorescence labeled Laminin Laminin

Biocompatible/Sterile Biocompatible/Sterile Prints accurate & precise patternsPrints accurate & precise patterns

High resolutionHigh resolution Cost-effective (<$500)Cost-effective (<$500)

Neural cell patterning1

Design AlternativesDesign Alternatives

Each alternative design consists of a Droplet-Each alternative design consists of a Droplet-Depositing-Device (DDD), Substrate-Positioning-Depositing-Device (DDD), Substrate-Positioning-Device (SPD), and Solution-Supplying-Device (SSD) Device (SPD), and Solution-Supplying-Device (SSD) that may or may not be the samethat may or may not be the same

1.1. X-Y Stepper Board with Stationary Print HeadX-Y Stepper Board with Stationary Print Head2.2. Modified Thermal Inkjet PrinterModified Thermal Inkjet Printer3.3. Modified Piezoelectric Inkjet PrinterModified Piezoelectric Inkjet Printer

Choose Best Design!Choose Best Design!

Technical ConsiderationsTechnical Considerations

Fibronectin SolutionFibronectin Solution 20ug/ml - 50ug/ml 20ug/ml - 50ug/ml

concentration in Hconcentration in H2200

Surface Polymer Surface Polymer Print on cover slides Print on cover slides

(150um thick)(150um thick) HydrophobicHydrophobic

Polystyrene, PDMS, Polystyrene, PDMS, PlasticPlastic

Increase cell patterning Increase cell patterning PEG, Pluronics, BSAPEG, Pluronics, BSA

Resolution- 635 dpiResolution- 635 dpi Diameter of droplet (2R): Diameter of droplet (2R):

20um20um Separation of drops (e): 20-Separation of drops (e): 20-

25um25um

Assessment of Design Alternatives Assessment of Design Alternatives

DDD: DDD: For a resolution of 635 dpi, it is necessary to minimize the printed For a resolution of 635 dpi, it is necessary to minimize the printed

droplet diameterdroplet diameterVelocity of Droplet (Ejection Frequency) Velocity of Droplet (Ejection Frequency) Surface tension and viscosity of solutionSurface tension and viscosity of solution

Clogging should be preventedClogging should be preventedPrevent ink spray phenomenonPrevent ink spray phenomenon

SPD:SPD: Movement of substrate should be minimized for high resolution Movement of substrate should be minimized for high resolution System should allow for uniquely dimensioned substrates to printed onSystem should allow for uniquely dimensioned substrates to printed on

SSD:SSD: Could make system robust for different viscosities and surface tensions Could make system robust for different viscosities and surface tensions

Different static pressuresDifferent static pressures Clogging should be preventedClogging should be prevented

Software handling for all three should be feasibleSoftware handling for all three should be feasible

Solution-Supplying-Device (SSD)Solution-Supplying-Device (SSD)

Ink CartridgeInk Cartridge Can’t control pressure where the ink leaves cartridge Can’t control pressure where the ink leaves cartridge It is difficult to decontaminate printer cartridgeIt is difficult to decontaminate printer cartridge Difficult to prevent protein loss in the porous membrane of Difficult to prevent protein loss in the porous membrane of

an ink cartridge (clogging)an ink cartridge (clogging)

Height Controlled ReservoirHeight Controlled Reservoir Variation in potential energy will yield different pressuresVariation in potential energy will yield different pressures Easily decontaminatedEasily decontaminated Clogging is not an issue Clogging is not an issue

Syringe PumpSyringe Pump Could be used to change pressure; because, small Could be used to change pressure; because, small

displacements yield large pressure changesdisplacements yield large pressure changes Clogging and Contamination can be easily resolved with a Clogging and Contamination can be easily resolved with a

syringe pump syringe pump

Syringe PumpSyringe Pump

X-Y Printing SystemX-Y Printing System

Automated X-Y Automated X-Y stepper board stepper board (SPD)(SPD) Stepper motorStepper motor

Stationary print Stationary print head (DDD)head (DDD) PiezoelectricPiezoelectric

Software is Software is availableavailable

System that uses stationary piezoelectric print head to deposit solution onto motorized X-Y stage1

X-Y Printing SystemX-Y Printing System

Piezoelectric head (DDD):Piezoelectric head (DDD): MicroFab:MicroFab:

Printed droplet diameter is 65um Printed droplet diameter is 65um (MIT(MIT11))

Separation of droplets is 10umSeparation of droplets is 10um Maximum ResolutionMaximum Resolution

is 339dpiis 339dpi Cost is $500Cost is $500

Can increase resolution by Can increase resolution by coupling the ejection of the coupling the ejection of the droplets with the movement of droplets with the movement of the boardthe board

Piezo drop-on-demand print head1

X-Y Printing SystemX-Y Printing System

•X-Y Movement (SPD)

Movement of X-Y stage relative to droplet velocity

X-Y Printing SystemX-Y Printing SystemModelModel PricePrice ResolutionResolution SpeedSpeedAB Tech AB Tech Mini X-Y Linear Motor Mini X-Y Linear Motor Positioning StagePositioning Stage

0.5 0.5 μμmm 100+ mm/s100+ mm/s

Newmark Systems inc.Newmark Systems inc.NLS4 SeriesNLS4 Series

>1350>1350 0.3 0.3 μμmm 12.5mm/s12.5mm/s

Rockwell AutomationRockwell Automation 0.1 0.1 μμmm 4mm/s4mm/s

Arrick Robotics XY-9Arrick Robotics XY-9 $750$750 .005" per .9 .005" per .9 degree stepdegree step

0.15m/s0.15m/s

X-Y Printing SystemX-Y Printing System

AdvantagesAdvantages Substrate-Dimension-Substrate-Dimension-

FreedomFreedom Software-ReadySoftware-Ready Viscosities and surface Viscosities and surface

tensions can be variedtensions can be varied

DisadvantagesDisadvantages CostlyCostly Low Resolution Low Resolution

(339dpi)(339dpi)Time consuming to Time consuming to couple the x-y board couple the x-y board with the printer headwith the printer head

Slow (5min/coverslip)Slow (5min/coverslip)

Thermal Inkjet PrintersThermal Inkjet PrintersHewlett-Packard Model 550CHewlett-Packard Model 550C Print head (DDD)Print head (DDD)

Thermal characteristicsThermal characteristics Boland et al.Boland et al.22

Printed droplet diameter 2umPrinted droplet diameter 2um

Separation of droplets is .25mmSeparation of droplets is .25mm

Maximum Resolution is 100dpiMaximum Resolution is 100dpi Software CapabilitiesSoftware Capabilities

Printer head element may only Printer head element may only change the ejection frequencychange the ejection frequency

Droplet size can not be changedDroplet size can not be changed Voltage through resistor is constantVoltage through resistor is constant

HP 550C3

Thermal Inkjet PrintersThermal Inkjet PrintersCan’t change the size of the droplet since Can’t change the size of the droplet since pressure in channel or the voltage pressure in channel or the voltage applied to the resistor is predetermined applied to the resistor is predetermined for the particular ink solutionfor the particular ink solution

The water-based ink becomes superheated (far above its normal boiling point) and finally reaches the critical temperature for bubble nucleation at around 280 °C. At this At this point no further heat is applied to the point no further heat is applied to the bubblebubble

Vapor bubble expands until all of the Vapor bubble expands until all of the heat stored in excess to the boiling heat stored in excess to the boiling point is used to convert liquid to point is used to convert liquid to vapor. Once the excess heat is vapor. Once the excess heat is removed, the bubble collapses on to removed, the bubble collapses on to the resistor, which is no longer being the resistor, which is no longer being heated.heated.

The bubble expansion and formation The bubble expansion and formation sequence occurs in 10us. Then there sequence occurs in 10us. Then there must be a minimum dwell time of must be a minimum dwell time of 100-500us to enable the channel to 100-500us to enable the channel to be refilled through capillary actionbe refilled through capillary action

Maximum ejection frequency is Maximum ejection frequency is 50kHz50kHz

Modified Thermal Inkjet PrinterModified Thermal Inkjet Printer

Caddy System (SPD)Caddy System (SPD) Re-design paper feed Re-design paper feed

mechanism of HP mechanism of HP printer printer

To allow 1mm+ thick To allow 1mm+ thick material to be inserted material to be inserted without rollingwithout rolling

Involves re-directing Involves re-directing feed and increasing feed and increasing height of rollersheight of rollers

Resolution may Resolution may decrease decrease

Printer with modified paper feed system4

Modified Thermal Inkjet PrinterModified Thermal Inkjet Printer

AdvantagesAdvantages Larger nozzle diameterLarger nozzle diameter

Mammalian Cells (100um)Mammalian Cells (100um) Fast (50,000 dots/min) Fast (50,000 dots/min)

DisadvantagesDisadvantages Thermal stress on Thermal stress on

biological materialsbiological materials Poor resolution Poor resolution

(100dpi)(100dpi) Need to design a Need to design a

caddy systemcaddy system Can’t control droplet Can’t control droplet

sizesize

Piezoelectric Inkjet PrinterPiezoelectric Inkjet PrinterEpson® Stylus R200Epson® Stylus R200

Print head (DDD)Print head (DDD) Built-in Caddy System (SPD)Built-in Caddy System (SPD)

Necessary to print on our substrateNecessary to print on our substrate Resolution of 5760x1440 dpi Resolution of 5760x1440 dpi

Maximum Diameter of Droplet Maximum Diameter of Droplet (4.5um)(4.5um)Diameter of droplet based on Diameter of droplet based on experimentsexperiments

Microencapsulation group achieved Microencapsulation group achieved a resolution of 254dpi (diameter of a resolution of 254dpi (diameter of droplet 100um)droplet 100um)

SoftwareSoftwareUse default Use default Modify voltage supply to actuatorModify voltage supply to actuator

CostCost$99$99

Epson Stylus R2005

Modified Piezoelectric PrinterModified Piezoelectric Printer

The size of the droplets is defined by the voltage applied to the deflection plate, the pulse duration, and the diameter of the orifice6

Modified Piezoelectric PrinterModified Piezoelectric Printer

AdvantagesAdvantages Caddy-ready systemCaddy-ready system High resolutionHigh resolution

Greater than 1200dpiGreater than 1200dpi77 Can vary pressure in Can vary pressure in

channelchannelMight be necessary for Might be necessary for solutions of different solutions of different viscosities and surface viscosities and surface tensions tensions

DisadvantagesDisadvantages Difficult to modify Difficult to modify

software software EPROM based EPROM based Microprocessor to work Microprocessor to work in conjunction with in conjunction with different power supply different power supply settingssettings

Ink SolutionInk SolutionHow do we increase the How do we increase the resolution?resolution?

Current inks have a Current inks have a surface tension of 3surface tension of 33.3.5 5 dyn/cm (0.0335N/m) dyn/cm (0.0335N/m) and a viscosity of 7.4 and a viscosity of 7.4 cP at 20 ◦CcP at 20 ◦C

The surface tension of The surface tension of water (73 dyne/cm) water (73 dyne/cm) would increase the would increase the force required to force required to deform dropletdeform droplet

Leading to an Leading to an increase in resolutionincrease in resolution

Viscosity of water Viscosity of water (0.89cp) would (0.89cp) would decrease the time decrease the time required to wet the required to wet the surfacesurface

Leading to a Leading to a decrease in decrease in resolutionresolution

Experimental ProtocolExperimental Protocol

Hypothesis:Hypothesis: By increasing the concentration of proteins in our By increasing the concentration of proteins in our

solution, the viscosity of our ink will increase; hence, solution, the viscosity of our ink will increase; hence, we will achieve higher resolutionwe will achieve higher resolution

By increasing the surface tension, we will prevent By increasing the surface tension, we will prevent wetting of our substratewetting of our substrate

A high surface tension means low attraction and a low A high surface tension means low attraction and a low surface tension means a high degree of attraction surface tension means a high degree of attraction

Image printed solution of fluorescent collagen Image printed solution of fluorescent collagen (type I)(type I) fluorescence microscopy fluorescence microscopy

Best Design SolutionBest Design Solution Modified Piezoelectric Printer Modified Piezoelectric Printer

Droplet-Depositing-Device (DDD) Droplet-Depositing-Device (DDD) High Resolution (High Resolution (5760x1440dpi)5760x1440dpi) Droplet Droplet

Size,Frequency Size,Frequency

Substrate-Positioning-Device (SPD)Substrate-Positioning-Device (SPD) Built in Caddy SystemBuilt in Caddy System

Solution-Supplying-Device (SSD)Solution-Supplying-Device (SSD) Syringe Pump Syringe Pump

SoftwareSoftware Easily Modified Easily Modified Versatile and Flexible cell patterning Versatile and Flexible cell patterning Epson Stylus R2005

Best Design SolutionBest Design Solution Modified Piezoelectric PrinterModified Piezoelectric Printer

QFD Analysis QFD Analysis Print Head (121.5) Print Head (121.5)

Easily ModifiedEasily Modified

ResolutionResolution Droplet SizeDroplet Size FrequencyFrequency

Low Cost (105) Low Cost (105) Print Head Print Head

Movement System Movement System

ReferencesReferences

1.1. Sanjana NE, et.al. A fast flexible ink-jet method for patterning dissociated neurons Sanjana NE, et.al. A fast flexible ink-jet method for patterning dissociated neurons in culture. J Neurosci Meth (2004); 136: 151-163.in culture. J Neurosci Meth (2004); 136: 151-163.

2.2. Boland T, et al. Cell and organ printing 1: protein and cell printers.Boland T, et al. Cell and organ printing 1: protein and cell printers.Anat Rec A Discov Mol Cell Evol Biol. 2003 Jun;272(2):491-6. Anat Rec A Discov Mol Cell Evol Biol. 2003 Jun;272(2):491-6.

3.3. ““HP 550C.” PC Universe (2005). <http://www.pcuniverse.hpsupplieslink.com>. 16 HP 550C.” PC Universe (2005). <http://www.pcuniverse.hpsupplieslink.com>. 16 Nov. 2005.Nov. 2005.

4.4. ““An idiot's guide to making a CD printer from an Epson 640.” VideoHelp.com An idiot's guide to making a CD printer from an Epson 640.” VideoHelp.com (2005) <http://www.videohelp.com/forum/viewtopic.php?t=177053> 15 Nov. 2005.(2005) <http://www.videohelp.com/forum/viewtopic.php?t=177053> 15 Nov. 2005.

5.5. ““Epson Stylus R200.” Epson America, Inc (2005). <http://www.epson.com>. 16 Epson Stylus R200.” Epson America, Inc (2005). <http://www.epson.com>. 16 Nov. 2005.Nov. 2005.

6.6. US Patent Office <http://www.uspatent.org>US Patent Office <http://www.uspatent.org>

7.7. Setti L et al., An amperometric glucose biosensor prototype fabricated by thermal Setti L et al., An amperometric glucose biosensor prototype fabricated by thermal ink jet printing. Biosen & Bioelec 20 (2005) 2019-2026. ink jet printing. Biosen & Bioelec 20 (2005) 2019-2026.