Molecular and Cellular applicationsewh.ieee.org/r1/syracuse/EMBSWeb/Archives/Events/... · 2009. 5....
Transcript of Molecular and Cellular applicationsewh.ieee.org/r1/syracuse/EMBSWeb/Archives/Events/... · 2009. 5....
Silicon nanomembranes (pnc-Si): Molecular and Cellular
applications James L. McGrathBiomedical EngineeringUniversity of RochesterNanomembrane Research Group
EMBS @ SY!CUSE UNIVERSITY APRIL 23, 2009
Friday, May 22, 2009
NRG 2009
Silicon Nanomembranes(pnc-Si - porous nanocrystalline silicon)
Thinner = faster transport and less sample loss
A. van den Berg and M. Wessling, Nature 445, 726, 2007
15nm thick5-80 nm pores
Friday, May 22, 2009
NRG 2009
Chris Striemer*
Bioengineeing Group Materials Group
Tom Gaborski*
Jess Snyder (Protein separations)Anant Agrawal (Cellular studies)Barrett Nehilla (Cell applications)Henry Chung (Microfluidics)Crowe, Hoffman, Summers (undergraduates)
Dave Fang (Material development)Maryna Kavalenka (Air permeability)
Jim McGrath Philippe Fauchet
SiMPore Inc.*
JP DesormeauxKarl Reisig
Nakul Nataraj
Rick RichmondJamie RoussieFunding
NSF, NIH, CSTI, J&J, NYSTAR, CEIS
Collaborations
Shigeru Amemiya, PITT
Nanomembrane Research Group
RIT: SMFL
Bill Bernhard, URMC
Friday, May 22, 2009
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Outline
Overview of pnc-Si
Transport
Separations
Cell Culture
Other Applications
Friday, May 22, 2009
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Fabrication
Striemer, C.S, et al. 2007, Nature, 445:749-53
Friday, May 22, 2009
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Fabrication RAPID = SECONDS
Striemer, C.S, et al. 2007, Nature, 445:749-53
Friday, May 22, 2009
NRG 2009
Fabrication RAPID = SECONDS
Striemer, C.S, et al. 2007, Nature, 445:749-53
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
physical vapor deposition
6
frontside
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
physical vapor deposition
6
frontsiderf magnetron sputtering
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
physical vapor deposition
6
20 nm sputtered SiO2
15 nm α-Si
frontsiderf magnetron sputtering
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
physical vapor deposition
6
20 nm sputtered SiO2
15 nm α-Si
frontsiderf magnetron sputtering
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
physical vapor deposition
6
20 nm sputtered SiO2
15 nm α-Si
frontsiderf magnetron sputtering
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
physical vapor deposition
6
20 nm sputtered SiO2
15 nm α-Si
frontside
amorphous Si
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
rapid thermal anneal
7
20 nm sputtered SiO2
15 nm α-Si
amorphous Si
frontside
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
20 nm sputtered SiO2
15 nm pnc-Si
rapid thermal anneal
7
700 °C – 1000 °C < 5 minpnc-Si
frontside
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
variety of formats
8
Friday, May 22, 2009
NRG 2009
Sharp Cut-offs and Tunable Pore Sizes
T = 715C
T = 729C
Striemer, C.S, et al. 2007, Nature, 445:749-53
NANOCRYSTALS
NANOPORES
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
pore size & density – rtp temperature
10
20 nm sputtered SiO2
15 nm pnc-Si
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
pore size & density – rtp temperature
10
700 C
20 nm sputtered SiO2
15 nm pnc-Si
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
pore size & density – rtp temperature
10
800 C
20 nm sputtered SiO2
15 nm pnc-Si
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
pore size & density – rtp temperature
10
850 C
20 nm sputtered SiO2
15 nm pnc-Si
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
pore size & density – rtp temperature
10
1000 C
20 nm sputtered SiO2
15 nm pnc-Si
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
pore size & density – rtp temperature
10
1100 C
20 nm sputtered SiO2
15 nm pnc-Si
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
controlling morphology – thickness
11
15 n
m p
nc-S
i
Friday, May 22, 2009
Nanomembrane Research Group [NRG]
controlling morphology – thickness
11
15 n
m p
nc-S
i30
nm
pnc
-Si
Friday, May 22, 2009
NRG 2009
Mechanically Robust
200 µm
Striemer, C.S, et al. 2007, Nature, 445:749-53
Friday, May 22, 2009
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Mechanically Robust
200 µm
15 PSI
Striemer, C.S, et al. 2007, Nature, 445:749-53
Friday, May 22, 2009
NRG 2009
Mechanically Robust
200 µm
15 PSI 0 PSI
Striemer, C.S, et al. 2007, Nature, 445:749-53
Friday, May 22, 2009
NRG 2009
Key Biological Sizes
smallsolutes
1 nm 10 nm 1 um
proteins virus bacteria
10 um
eukaryoticcells
Friday, May 22, 2009
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Key Biological Sizes
smallsolutes
1 nm 10 nm 1 um
proteins virus bacteria
10 um
eukaryoticcells
PNC-SI (5-80 NM)
Friday, May 22, 2009
NRG 2009
Key Biological Sizes
smallsolutes
1 nm 10 nm 1 um
proteins virus bacteria
10 um
eukaryoticcells
PNC-SI (5-80 NM)@ 0.1%-15%
Friday, May 22, 2009
NRG 2009
3 L fluorescent mixture (PBS)
glass co
versl
ipPBS buffer
15 nm thicknc-Si membrane
Species 1 -
50 m silica spacer
Species 2 -
~ 500 mmicroscope
experimentalfield of view
t = 0 min
100 m
Striemer, C.S, et al. 2007, Nature, 445:749-53
Size Based Separation
Friday, May 22, 2009
NRG 2009Striemer, C.S, et al. 2007, Nature, 445:749-53
Size Based Separation
Friday, May 22, 2009
NRG 2009Striemer, C.S, et al. 2007, Nature, 445:749-53
Size Based Separation
Friday, May 22, 2009
NRG 2009Striemer, C.S, et al. 2007, Nature, 445:749-53
Size Based Separation
Friday, May 22, 2009
NRG 2009
Time, min0 1 2 3 4 5 6 7
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
Flu
ores
cenc
e in
tens
ity, a
.u.
BSA
Alexa dye
1 nm
7 nm
MEMBRANE A
Striemer, C.S, et al. 2007, Nature, 445:749-53
Size Based Separation
Friday, May 22, 2009
NRG 2009
Time, min0 1 2 3 4 5 6 7
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
Flu
ores
cenc
e in
tens
ity, a
.u.
BSA
Alexa dye
1 nm
7 nm
MEMBRANE A
Time, min0 1 2 3 4 5 6 7
BSA
IgG
0.4
0.3
0.2
0.1
0
Flu
ores
cenc
e in
tens
ity, a
.u.
7 nm
14 nm
MEMBRANE B
Striemer, C.S, et al. 2007, Nature, 445:749-53
Size Based Separation
Friday, May 22, 2009
NRG 2009
Alexa dye - charge = 2-
0 1 2 3 4 5 6 7 8 9 10
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0
Time, min
Flu
ores
cenc
e in
tens
ity, a
.u.
Amine - DI H2O
Oxide - 100 mM NaCl
Oxide - DI H2O
--------
--------
------
-
-
---
--
--
-- -
Striemer, C.S, et al. 2007, Nature, 445:749-53
Charge Based Separation
Friday, May 22, 2009
NRG 2009
Alexa dye - charge = 2-
0 1 2 3 4 5 6 7 8 9 10
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0
Time, min
Flu
ores
cenc
e in
tens
ity, a
.u.
Amine - DI H2O
Oxide - 100 mM NaCl
Oxide - DI H2O
-
-
---
--
--
-- -
Striemer, C.S, et al. 2007, Nature, 445:749-53
Charge Based Separation
Friday, May 22, 2009
NRG 2009
Alexa dye - charge = 2-
0 1 2 3 4 5 6 7 8 9 10
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0
Time, min
Flu
ores
cenc
e in
tens
ity, a
.u.
Amine - DI H2O
Oxide - 100 mM NaCl
Oxide - DI H2O
-
-
---
--
--
-- -
++++++++
++++++++
++++++
Striemer, C.S, et al. 2007, Nature, 445:749-53
Charge Based Separation
Friday, May 22, 2009
NRG 2009
Alexa dye - charge = 2-
0 1 2 3 4 5 6 7 8 9 10
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0
Time, min
Flu
ores
cenc
e in
tens
ity, a
.u.
Amine - DI H2O
Oxide - 100 mM NaCl
Oxide - DI H2O
-
-
---
--
--
-- -
++++++++
++++++++
++++++
-
-
---
--
--
-- -
Striemer, C.S, et al. 2007, Nature, 445:749-53
Charge Based Separation
Friday, May 22, 2009
NRG 2009
Alexa dye - charge = 2-
0 1 2 3 4 5 6 7 8 9 10
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0
Time, min
Flu
ores
cenc
e in
tens
ity, a
.u.
Amine - DI H2O
Oxide - 100 mM NaCl
Oxide - DI H2O
-
-
---
--
--
-- -
-
-
---
--
--
-- -
Striemer, C.S, et al. 2007, Nature, 445:749-53
Charge Based Separation
Friday, May 22, 2009
NRG 2009
Alexa dye - charge = 2-
0 1 2 3 4 5 6 7 8 9 10
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0
Time, min
Flu
ores
cenc
e in
tens
ity, a
.u.
Amine - DI H2O
Oxide - 100 mM NaCl
Oxide - DI H2O
-
-
---
--
--
-- -
Striemer, C.S, et al. 2007, Nature, 445:749-53
Charge Based Separation
Friday, May 22, 2009
NRG 2009
Alexa dye - charge = 2-
0 1 2 3 4 5 6 7 8 9 10
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0
Time, min
Flu
ores
cenc
e in
tens
ity, a
.u.
Amine - DI H2O
Oxide - 100 mM NaCl
Oxide - DI H2O
-
-
---
--
--
-- -
NaClStriemer, C.S, et al. 2007, Nature, 445:749-53
Charge Based Separation
Friday, May 22, 2009
NRG 2009
Alexa dye - charge = 2-
0 1 2 3 4 5 6 7 8 9 10
0.45
0.40
0.35
0.30
0.25
0.20
0.15
0.10
0.05
0
Time, min
Flu
ores
cenc
e in
tens
ity, a
.u.
Amine - DI H2O
Oxide - 100 mM NaCl
Oxide - DI H2O
-
-
---
--
--
-- -
NaClStriemer, C.S, et al. 2007, Nature, 445:749-53
-
-
---
--
--
-- -
Charge Based Separation
Friday, May 22, 2009
NRG 2009
Scalable, flexible, economical fabrication
Friday, May 22, 2009
NRG 2009
Production Flow
PATTERNING DEPOSITION THERMAL ANNEALING
ETCHING TESTING & APPLICATIONS
PAST
Friday, May 22, 2009
NRG 2009
Production Flow
PATTERNING DEPOSITION THERMAL ANNEALING
ETCHING TESTING & APPLICATIONS
PAST
NEAR FUTURE
ALL FABRICATION TESTING & APPLICATIONS
Friday, May 22, 2009
NRG 2009
Production Flow
PATTERNING DEPOSITION THERMAL ANNEALING
ETCHING TESTING & APPLICATIONS
PAST
NEAR FUTURE
ALL FABRICATION TESTING & APPLICATIONS
MADE POSSIBLE BY PURCHASE OF NEW SYSTEMS
• SURFACE PREP• ANNEALING • DEPOSITION
Friday, May 22, 2009
NRG 2009
UR’s Nanoscience Center
Friday, May 22, 2009
NRG 2009
UR’s Nanoscience Center
MCGRATH LAB
Friday, May 22, 2009
NRG 2009
UR’s Nanoscience Center
MCGRATH LAB
SURFACE PREP, DEPOSITION AND
ANNEALING
Friday, May 22, 2009
NRG 2009
UR’s Nanoscience Center
MCGRATH LAB
SURFACE PREP, DEPOSITION AND
ANNEALING
LITHOGRAPHY
Friday, May 22, 2009
NRG 2009
Manufacturing Possibilities and Challenges
4” wafer
CURRENTLY: 68 INSERTS PER WAFER
Friday, May 22, 2009
NRG 2009
Manufacturing Possibilities and Challenges
4” wafer
CURRENTLY: 68 INSERTS PER WAFER
6” wafer
> 500 INSERTS PER WAFER
Friday, May 22, 2009
NRG 2009
Manufacturing Possibilities and Challenges
0.004 CM2
0.33 CM2
Friday, May 22, 2009
NRG 2009
Manufacturing Possibilities and Challenges
0.004 CM2
0.33 CM2
• LESS THAN 1% OF AREA IS CURRENTLY ACTIVE
• MADE NECESSARY BY ETCHING, MECHANICS, AND DEFECT FREQUENCY
Friday, May 22, 2009
NRG 2009
Manufacturing Possibilities and Challenges
0.004 CM2
0.33 CM2
• LESS THAN 1% OF AREA IS CURRENTLY ACTIVE
• MADE NECESSARY BY ETCHING, MECHANICS, AND DEFECT FREQUENCY
200 microns
Friday, May 22, 2009
NRG 2009
Ultrathin SiN membranesTong, et al. (2004). Silicon Nitride Nanosieve Membrane. Nano Letters 4, 283-287
SiN - 10 nm thick but elaborate and impractical
No demonstrated separations
Appreciated all the potential and issues (air flow, water permeability, and mechanics)
Friday, May 22, 2009
NRG 2009
10 µm-diameter tip Ru(NH3)63+ in 0.1 M KCl
High Diffusive PermeabilityInstrinsic permeability of 5.2 x 10-2 cm/s measured using Scanning Electrochemical Microscopy.
Experimental results & pore histograms are consistent with theory that neglects pore resistance
Between 2-3 orders higher than small molecule diffusion permeability of reconstituted cellulose or PES.
Kim et al. 2008, JACS 130:4230-4231
Shigeru Amemiya
Friday, May 22, 2009
NRG 2009
High Diffusive PermeabilityInstrinsic permeability of 5.2 x 10-2 cm/s measured using Scanning Electrochemical Microscopy.
Experimental results & pore histograms are consistent with theory that neglects pore resistance
Between 2-3 orders higher than small molecule diffusion permeability of reconstituted cellulose or PES.
Kim et al. 2008, JACS 130:4230-4231
Shigeru Amemiya
Friday, May 22, 2009
NRG 2009
High Diffusive PermeabilityInstrinsic permeability of 5.2 x 10-2 cm/s measured using Scanning Electrochemical Microscopy.
Experimental results & pore histograms are consistent with theory that neglects pore resistance
Between 2-3 orders higher than small molecule diffusion permeability of reconstituted cellulose or PES.
Kim et al. 2008, JACS 130:4230-4231
Shigeru Amemiya
Friday, May 22, 2009
NRG 2009
High Diffusive PermeabilityInstrinsic permeability of 5.2 x 10-2 cm/s measured using Scanning Electrochemical Microscopy.
Experimental results & pore histograms are consistent with theory that neglects pore resistance
Between 2-3 orders higher than small molecule diffusion permeability of reconstituted cellulose or PES.
Kim et al. 2008, JACS 130:4230-4231
Shigeru Amemiya
k = 2DNr
Friday, May 22, 2009
NRG 2008
Higher Permeability = Smaller ...
For current devices: active area is 1.8 m2 and transmembrane water flow is ~10 ml/min @ 3 psi.
1000x improved permeability would require only 18 cm2. So 10 dime-sized membranes with mostly (~80%) active area could support both transmembrane flow and match dialysis in the same period of time.
Friday, May 22, 2009
NRG 2008
Higher Permeability = Smaller ...
For current devices: active area is 1.8 m2 and transmembrane water flow is ~10 ml/min @ 3 psi.
1000x improved permeability would require only 18 cm2. So 10 dime-sized membranes with mostly (~80%) active area could support both transmembrane flow and match dialysis in the same period of time.
Friday, May 22, 2009
NRG 2008
… or faster?Alternatively a 6” wafer of mostly active membrane could achieve the same dialysis in 1/10th of the time.
Friday, May 22, 2009
NRG 2008
… or faster?Alternatively a 6” wafer of mostly active membrane could achieve the same dialysis in 1/10th of the time.
THE MATERIAL PROVIDES THE POTENTIAL
REALIZING THIS POTENTIAL IS AN ENGINEERING CHALLENGE
Friday, May 22, 2009
NRG 2009
Cellular Transwell DevicesCO-CULTURE
Friday, May 22, 2009
NRG 2009
Cellular Transwell DevicesCO-CULTURE
DRUG PERMEATION
Friday, May 22, 2009
NRG 2009
Microfluidics
Friday, May 22, 2009
NRG 2009
ConclusionsPnc-Si is a new ultrathin nanoporous membrane material. Small membranes can be manufactured on a large scale and incorporated into practical separation devices.
Primary application is to small scale separation of biologicals
High air and liquid permeabilities w/ demonstrated ability to fractionate proteins, nanoparticles, etc.
Viable as a cell-culture substrate. Cell behavior on membranes is normal.
Microfluidics, arrayed membranes for screening applications, electrokinetics, and more …
Friday, May 22, 2009