microfluidics vs. self-assembly

... talk given at the Institut Pierre-Gilles de Gennes, Paris (October 11, 2013)

1. microfluidics vs self assembly microfluidics vs. self-assembly Andreas Manz KIST Europe, Saarbrücken, Germany KIST Seoul South Korea KIST Seoul, South Korea Mechatronics, Saarland University, Germany
2. … questions like: “how is a butterfly wing manufactured?” wing manufactured? • microstructure, nanostructure, colour t bl t i l ( hiti ) t li • stable material (chitin), not alive • reproducibility • ease of manufacturing • low cost • … and what is the blueprint for it?
3. … questions like: “how is a butterfly wing manufactured?” wing manufactured? d h t i th bl i t f it? •… and what is the blueprint for it? • … how to get from molecular biology to structure? • … how to get discrete size, structure • … how to engineer by self assembly? g y y
4. … questions like: “how is a butterfly wing manufactured?” wing manufactured? d h t i th bl i t f it? •… and what is the blueprint for it? • … how to get from molecular biology to structure? • … how to get discrete size, structure • all 3 have identical genome g
5. … questions like: “how is a butterfly wing manufactured?” wing manufactured? Morphidae, 170mm smallest feature 100nm
6. … questions like: “how is a butterfly wing manufactured?” wing manufactured? Morphidae, 170mm smallest feature 100nm
7. … questions like: “how is a butterfly wing manufactured?” wing manufactured? Morphidae, 170mm 100nm
8. … note: I am not yet speaking of engineering a microsystem like this engineering a microsystem like this…. Syrphidae 7mm Syrphidae, 7mm
9. what is “lab on chip” technology? • device made from a substrate dev ce de o subs e • using clean room technology • target: chemistry biology medical use • target: chemistry, biology, medical use • containing channels, reactors etc t i d t t h t t • may contain detectors, heaters, etc.
10. why is it difficult? l t h l i i • clean room technology is expensive • labour intensive • mistakes in layout difficult to correct • (take my example…) ( y p )
12. ciba-geigy, basel switzerland (now novartis solvias) (now novartis, solvias) 1988-1996
13. ciba-geigy, basel switzerland (now novartis solvias) (now novartis, solvias) 1988-1996
14. imperial college london 1996 2006 1996-2006
15. imperial college london 1996 2006 1996-2006
16. imperial college london 1996 2006 1996-2006
17. isas, dortmund germany 2003 2008 2003-2008
18. glass‐glass substrate materials pmma‐pmma glass glass pdms‐pdms pdms‐glass glass silicon glass‐glass silicon‐silicon glass‐silicon silicon‐silicon pmma‐pmma glass gold glass glass‐silicon glass‐gold‐glass glass‐laminate‐glass glass‐silicon‐glass d l l i l d d pdms‐glass ordyl multi‐layer glass pdms‐copper pdms‐silicon pdms‐pdms pdms‐silicon‐pdms quartz‐quartz • substrate materials used
19. integrated features heaters porous membrane nothing t l l t d heaters g metal electrodes heaters porous membrane slit array outside metal electrodes slit array outside liquid membrane planar waveguides x‐ray source nothing t sensor phase quides • integrated featurs, like metal electrodes, heaters, membranes etc
20. topology of channels binary branching structure non‐binary branching well tree, spider single channel central bed, ch around single channel 1 loop central ch array, tree central bed, ch around tree, spider central ch array, tree binary branching structure non‐binary branching well central chamber, frit, tree single channel 1 loop network central chamber, single ch • topology • spider, tree, loop, network, etc p p
21. interfacing type flat plastic plates large holes, thick pdms cover large holes, thick glass cover eppendorf pipets open eppendorf pipets, open flat metal plates eppendorf pipets, open fused silica tubing don't know, not used plastic tubing ‐ glue fused silica tubing plastic tubing ‐ glue flat metal plates large holes, thick glass cover flat plastic plates large holes thick pdms cover don't know, not used large holes, thick pdms cover • interfacing type • (“chip to world interface”) p
22. application area pumping sample prep basics application p p g separation d t ti separation biology detection reaction biology reaction basics application pumping pumping sample prep detection • what is the chip used for?
23. commercializations commercial commercial attempt no attempt attempt no attempt • How many chips were in direct line to commercialization?
24. take the best example ill l t h i • capillary electrophoresis • scaling: 100x smaller (length) • time to result: < 10,000x faster • targets RNA or DNA analysis g y
25. • capillary electrophoresis, flow injection, electrochemical detection • glass – glass chip, design 1989, fab 1989 mettler imt switzerland g g p g • manz, fettinger, lüdi, widmer, svs bulletin 5, 4-10, 1990
26. • capillary electrophoresis, injection, electrochemical detection • glass – glass chip, design 1989, fab 1989 mettler imt switzerland g g p g • manz, harrison, fettinger, verpoorte, lüdi, widmer, proc. transducers 1991 san francisco, 939-941, 1991
27. • capillary electrophoresis, injection • glass – glass chip, design 1992, fab 1992 mettler imt switzerland g g p g • effenhauser, manz, widmer, anal.chem. 65, 2637-2642, 1993
28. • synchronised cyclic capillary electrophoresis, injection • glass – glass chip, design 1992, fab 1992 mettler imt switzerland g g p g • burggraf, manz, effenhauser, verpoorte, de rooij, widmer, j.high resolut.chromatogr. 16, 594-596, 1993
29. • 2d capillary electrophoresis, injection • quartz – quartz chip, design 1996, fab 1997 imm mainz germany q q p g g y • becker, lowack, manz, j.micromech.microeng. 8, 24-28, 1998
30. • capillary electrophoresis, parallel processing, injection • glass – glass chip, design 1996, fab 1996 caliper ltd. california usa g g p g p • manz, becker, proc. transducers 1997 chicago, 915-918, 1997
31. • capillary electrophoresis, parallel processing, injection • glass – glass chip, design 1996, fab 1996 caliper ltd. california usa g g p g p • manz, becker, proc. transducers 1997 chicago, 915-918, 1997
32. • 2d capillary electrophoresis, injection • glass – glass chip, design 1996, fab 1996 caliper ltd. california usa g g p g p • manz, bousse, unpublished (patent filing 2002)
33. … and some results • proof of principle p oo o p c p e • high speed separation • commercial product • commercial product • market needs just 2x faster electrophoresis ( h di i ti !) • (…. how disappointing!)
34. capillary electrophoresis results capillary electrophoresis results flow injection results electrochemical results missing • capillary electrophoresis, flow injection, electrochemical detection • manz, fettinger, lüdi, widmer, svs bulletin 5, 4-10, 1990 g
35. • capillary electrophoresis, injection • manz, harrison, fettinger, verpoorte, lüdi, widmer, proc. transducers g p p 1991 san francisco, 939-941, 1991
36. • capillary electrophoresis, small molecules, fluorescence • manz, harrison, verpoorte, fettinger, paulus, lüdi, widmer, p g p j.chromatogr. 593, 253-258, 1992 • harrison, manz, fan, lüdi, widmer, anal.chem. 64, 1926-1932, 1992
37. • capillary electrophoresis, amino acids • effenhauser, manz, widmer, anal.chem. 65, 2637-2642, 1993
38. • capillary electrophoresis, phosphorothioate oligomers • effenhauser, paulus, manz, widmer, anal.chem. 66, 2949-2953, 1994 p
39. • capillary electrophoresis, fractionation, phosphorothioate oligomers • effenhauser, manz, widmer, anal.chem. 67, 2284-2287, 1995
40. • synchronised cyclic capillary electrophoresis, injection • burggraf, manz, effenhauser, verpoorte, de rooij, widmer, j.high resolut.chromatogr. 16, 594-596, 1993 • burggraf, manz, verpoorte, effenhauser , widmer, de rooij, sens actuators b20 103-110 1994
41. • synchronised cyclic MEKC or capillary electrophoresis, injection • von heeren, verpoorte, manz, thormann, anal.chem. 68, 2044-2053, 1996
42. • synchronised cyclic MEKC or CE, theophylline immunoassay • von heeren, verpoorte, manz, thormann, anal.chem. 68, 2044-2053, 1996
43. • synchronised cyclic MEKC, human urine, derivatized with FITC • von heeren, verpoorte, manz, thormann, anal.chem. 68, 2044-2053, 1996
44. • synchronised cyclic gel electrophoresis, amino acids • von heeren, verpoorte, manz, thormann, j.microcolumn separations 8, 373-381, 1996
45. • synchronised cyclic gel electrophoresis, phosphorothioate oligonucleotides T2 -T10 • von heeren, verpoorte, manz, thormann, j.microcolumn separations 8, 373-381, 1996
46. • label-free carbohydrate detection, holographic optical element • burggraf, krattiger, de mello, de rooij, manz, the analyst 123, 1443- 1447, 1998
47. capillary electrophoresis results missing • 2d capillary electrophoresis, injection • becker, lowack, manz, j.micromech.microeng. 8, 24-28, 1998 j g
48. • capillary electrophoresis, parallel processing, injection • manz, becker, proc. transducers 1997 chicago, 915-918, 1997 p g
49. • capillary electrophoresis, parallel processing, injection • manz, becker, proc. transducers 1997 chicago, 915-918, 1997 p g
50. • capillary electrophoresis, parallel processing, injection • manz, becker, proc. transducers 1997 chicago, 915-918, 1997 p g
51. • capillary electrophoresis, parallel processing, injection • manz, becker, proc. transducers 1997 chicago, 915-918, 1997 p g
52. • capillary electrophoresis, injection • manz, bousse, unpublished p • presented first at ISPPA, tomakomai, japan 1998
53. • capillary electrophoresis, injection • manz, bousse, unpublished p • presented first at ISPPA, tomakomai, japan 1998
54. • capillary electrophoresis, injection • manz, bousse, unpublished p • presented first at ISPPA, tomakomai, japan 1998
55. • capillary electrophoresis, injection • manz, bousse, unpublished p • presented first at ISPPA, tomakomai, japan 1998
56. … everything quite an effort … • seeking alternatives see g e ves • particulary for manufacturing • looking at examples in nature • looking at examples in nature • structured approach lf bl • self assembly
57. starting very simple • 3 phase system • “self assembly” energy driven • self assembly , energy driven • a droplet (just that)
58. Virtual Reaction Chamber Key properties – Water-based sample encapsulated by oil – (RT) PCR conducted on a PCR Oi glass cover slip – Micromachined heater/sensor Sample Oi l B are separated from the sample – Cover slip is disposable – Small sample volume makes system very fast Mirror reflection
59. VRC details LENGTH HEATER SENSOR Key properties – VRC with glass placed on a LENGTH LINK SENSOR micromachined silicon – Heater integrated with LINK temperature sensor – Heating rate: thermal mass, available power with PID control – Cooling rate:  (thermal time constant) H T G P G H    ; 
60. Ultrafast VRC, 650 K/s!!! 180 700 From room temperature to 150 oC in 0.2 s!! 140 160 600 V) 100 120 ature (oC) cence (mV 60 80 Tempera 500 Fluoresc 20 40 400 0 5 10 15 20 Time (s)
61. Avian Influenza Virus Detection by RT-PCR Key properties • SYBR-Green Real-Time RT- PCR 0 .6 2 ature (V) • Melting Curve Analysis • 8 minutes for RNA detection 0 1 Tempera 8 utes o N detect o 0 .3 -2 -1 e (V) V irusD etected H ot S tart P C R 100 150 10-2 cence (mV) uorescence (V/cycle) -3 uorescence V irus D etected R T 0 50 10-3 Fluoresc Differential Flu Critical Threshold 22.3 0 2 4 6 8 1 0 12 0 .0 -5 -4 Flu 0 10 20 30 40 10-4 Cycle Number T im e (m in)
62. Palm-sized PCR
63. sample preparation 1) disruption of spores by superheating for fast DNA extraction fast DNA extraction 2) protein and peptide decomposition by 2) protein and peptide decomposition by superheating 63
64. superheating solvent is at a temperature higher than boiling point without boiling! PCR Oi without boiling! Sample Oi l B experiment mirror reflection no boiling of aqueous solutions at 240 °C for more than 30 min!!! limited by thermal decomposition of surrounding oil y p g temperature x exposure time = applied energy 64
65. Bacillus spore disruption by superheating spores of bacteria are highly resistance against: - dryness y - toxic substances - other aggressive substances substances - aging - heat: dry: 150 °C ca. 1 h boiling: ca 5 h boiling: ca. 5 h electron microscope cross section of a spore of Bacillus electron microscope cross-section of a spore of Bacillus subtilis, showing the cortex and coat layers surrounding the core (dark central area). spore is 1.2 µm across. (Picture: S. Pankratz, Berkeley University of California) 65
66. B. subtilis purified spores microscope image of Bacillus subtilis spores after contrast staining (spores: blue) contrast staining (spores: blue) Z i A i 2 1500 ifi i 66 Zeiss Axiotron 2, 1500 magnification
67. B. subtilis purified spores after SUPERHEATING microscope image of Bacillus subtilis spores after contrast staining (spores: blue) contrast staining (spores: blue) Z i A i 2 1500 ifi i 67 Zeiss Axiotron 2, 1500 magnification
68. spore disruption destruction of spores by superheating 1 0 1 p o s itiv e c o n tro l n e g a tiv e c o n tro l 1 0 0 1 0 tensity s p o re s o lu tio n s p o re s a fte r p re tre a tm e n t s p o re s a fte r s u p e rh e a tin g 1 0 -1 1 0 scence int 1 0 -2 1 0 Fluore 0 5 1 0 1 5 2 0 2 5 3 0 3 5 4 0 1 0 C yc le N u m b e r 68
69. protein and peptide protein- and peptide- decomposition by superheating p y p g for peptide mass fingerprinting 69
70. start with “easy” samples: ACTH • adrenocorticotropic hormone (fragment 1-24) • molecular weight 2933.44 Da • ACTH is a biomarker for cellular stress, infections, cancer (metastases!), activates G proteins… 70
71. peptide decomposition by superheating 2628.2 60 70 80 90 100 Intensity 2932.645 No heating 49.0 640.8 1232.6 1824.4 2416.2 3008.0 Mass (m/z) 10 20 30 40 50 % 1467.313 g Mass (m/z) 5354.2 60 70 80 90 100 tensity 2932.6687 Superheating to 130 °C for 49 0 640 8 1232 6 1824 4 2416 2 3008 0 10 20 30 40 50 60 % In 2 1467.3292 360.343 2724.5750 1475.3208 978.5501 213.122 2885.6414 1635.0529 10 s 49.0 640.8 1232.6 1824.4 2416.2 3008.0 Mass (m/z) 2.0E+4 60 70 80 90 100 ensity 2932.6814 Superheating to 130 °C for 49 0 640 8 1232 6 1824 4 2416 2 3008 0 10 20 30 40 50 60 % Inte 2 1467.3354 2915.7068 2724.5742 1635.0637 379.0871 71.3642 2835.6299 978.8941 2884.6804 2682.5625 1363.2769 1539.3729 1498.2889 1691.1270 p g 20 s 71 49.0 640.8 1232.6 1824.4 2416.2 3008.0 Mass (m/z)
72. how about a challenge? • manufacture an object hi h h ld b h d which you can hold by hand • from smaller parts which you cannot hold by hand • by self assembly y y • by structured approach
73. Organelle Atoms Smooth muscle cell Molecule Atoms Chemical level At bi t f l l Cellular level Cells are made up of molecules. 1 2 Cardiovascular system Atoms combine to form molecules. Tissue level Smooth muscle tissue 3 system Tissues consist of similar types of cells. Blood vessel (organ) Heart Blood vessels Connective tissue Smooth muscle tissue Organ level Epithelial tissue 4 Organ level Organs are made up of different types of tissues. Organ system level Organism level 4 5 6 g y Organ systems consist of different organs that work together closely. Organism level The human organism is made up of many organ systems. 6
74. hierarchical assembly y
75. self assembly self assembly S. A. Stauth, C. J. Morris, and B. A. Parviz,in Evolvable Hardware 2004, Seattle, WA, 2004 Y H Jhang et al Organic Electronics Y.-H. Jhang et al., Organic Electronics, 13(10), pp. 1865-1872, 2012 K. Hosokawa, I. Shimoyama, and H. Miura, S & A t t A 57 117 125 1996 T. L. Breen et al., Science, 284, pp. 948-951, Sensors & Actuators A, 57, pp. 117-125, 1996 1999
76. self assembly y S. E. Chung et al., Nature Materials, 5, pp. 1147, 2008 S A St th d B A P i PNAS 103(38) C. Lin, Y. Liu, and H. Yan, biochemistry, 48(8), pp. 1663-1674, 2009 S. A. Stauth and B. A. Parviz, PNAS, 103(38), pp. 13922-13927, 2006
77. concept • the use of hard material • achievement of asymmetric pattern by logical sequence • morphology-based assembly (non chemical functionalization) ) • capillary force as driving force • tripods as building blocks • tripods as building blocks • assembly at fluidic interface
78. lateral capillary force
79. capillary force Surface Surface Air Su ace tension (γ) Su ace tension Meniscus Meniscus Tripod Water Contact angle (θc ) Water
80. capillary attraction As approaching each other the contact angle is decreased P. Singh et al., Soft Matter, 2010, 6, 4310-4325 As approaching each other, the contact angle is decreased and laterally attractive capillary force is increased
81. capillary attraction Capillary attraction between hydrophobic & floating material http://www.youtube.com/watch?v=TAY6RcJPEHY
82. size effect In order to increase Bond number, higher density, larger size, and weaker surface tension of floating material and medium are necessary necessary.
83. examples of LOGIC for self-assembly
84. B B’ can bind to examples of LOGIC for self-assembly
85. symmetric type • ANTHRACENE • Six different tripods are needed for this Six different tripods are needed for this assembly.
86. B A B A’ B A B B C B B C (upset patterns are !) A B C A’ B C same!) A B C B C B B C C B B C B’ B’ 2x C B B C C X and X’ are pairs p
87. C C’ C’ 2x 2x C 2x 2x
88. less symmetric type • PHENANTHRENE • 13 different tripods are needed for this 13 different tripods are needed for this assembly.
89. B B A B A’ B B B B C A’ B B C A A’ B D B D B D B B C B’ B’ C B B D B B 2x D B B D D
90. E E D’ D’ A E A’ D A B B C B B A A’ B D B D B E D B F D’ F ’ B’ B’ E’ B’ B B
91. F C C D’ F D F’ C C C D C C’ C’ D’
93. design and fabrication of assembling elements assembling elements
94. our choice • Tripod: Plastic (SU-8) • Interface: water/air • The dimension of tripods: L 500 μm Material Densit (g/cm3) Yo ng’s mod l s(GPa) • The dimension of tripods: L~ 500 μm Material Density (g/cm3) Young’s modulus(GPa) Silicon 2.33 130-188 SU-8 1.19 4.02 PDMS 0.965 0.0018 Polyimide 1.43 3.2
95. Fabrication Procedure of SU 8 Tripods 1 Omnicoat is used as releasing Fabrication Procedure of SU-8 Tripods 1. Omnicoat is used as releasing agent of SU-8 microstructure. 2. The stress of the structure should Coating omnicoat and SU-8 2050 on the wafer be minimized (RT curing, no sudden thermal-process) 3 Th t th th t i d 3. The way to gather the tripods without stacking each other is necessary Curing at RT and patterning necessary patterning Filtration for obtaining SU-8 tripods Releasing the tripods by dipping in the Remover PG
96. first design for tripods
97. first design for tripods
98. process flow process flow 1. Fabricated SU-8 pattern 2. Diced sample 3. Release of tripods from the wafer 4. Placement of the filter paper on the filter 5. Configuration of the filtration system with vacuum pump 6. Filtration 7. Washing with D.I. water 8. Vacuum- drying of the 9. Observation with microscope 10. The petridish with floating tripod thoroughly filter paper elements
99. fabricated tripods Fabricated SU-8 tripod on the wafer Released SU-8 tripod on the wafer
100. assembly results
101. ideal dimer formation
102. observed dimer formation
103. assembled tripods
104. t l ti rectangular tip case
105. elimination of local minimum elimination of local minimum - - Elimination of local Elimination of local Elimination of local Elimination of local minimum minimum A Elimination of local Elimination of local - - Elimination of local Elimination of local minimum minimum - - Round tip for Round tip for p p minimizing the minimizing the interacting area interacting area i i i i i i - - Sliding gradient Sliding gradient B
106. results (A type) results (A type)
107. results (B type) results (B type)
108. tripods tripods
109. tripods tripods
110. tripods tripods
111. tripods tripods
112. snapshots snapshots t=0 s t=0.25 s t=0.5 s t=0.75 s t=1 s t=0.8 s
113. smaller tripods smaller tripods Th tt ti f i t The attractive force is not strong enough to make them assembled because them assembled because the smaller size leads to smaller bond number and interfacial deformation
114. … lipid extrusions … li id t b l • lipid tubules • reproducible • um size • (lifetime limited) ( )
116. vesicle production PDMS p PDMS Si PDMS Si PDMS
117. vesicle production p PDMS Si PDMS 100 µm 2 µm
118. vesicle production p flow direction 100 µm 100 µm side view side view fluorescence image
119. formation of P.S.Dittrich, M.Heule, P.Renaud, A.Manz Lab Chip 6 488-493 (2006) formation of vesicle tubes Lab Chip 6, 488 493 (2006)
120. Formation of helices 50 µm 50 µm P.S.Dittrich, M.Heule, P.Renaud, A.Manz Lab Chip 6, 488-493 (2006)
121. … ongoing work … t t i • spontaneous extrusions • parallel extrusions • tubing, cilia, large surface materials • soft materials
124. CONCLUSION CONCLUSION
125. CONCLUSION CONCLUSION • biomimetic microfabrication may be very interesting for manufacturing ill i i d i l • still curiosity driven, very early stage • concepts for selective hierarchical • concepts for selective hierarchical assembly needed y
126. k l d t acknowledgement Leon Abelmann, PhD, Professor Pavel Neuzil, PhD Matthias Altmeyer PhD Matthias Altmeyer, PhD Eric Castro, PhD Adam Pribylka V Al id In Korea: Vanessa Almeida Per Arvid Loethman Seung Jae Lee Tae Song Kim, KIST Seoul, Korea Seungwon Jung KIST Seoul Korea Mi Jang Himani Sharma Jukyung Park Seungwon Jung , KIST Seoul, Korea Min Cheol Park , KIST Seoul, Korea Pavithra Sukumar , KIST Seoul, Korea Christian Ahrberg Tim Mehlhorn Camila Madeira Campos Ca a ade a Ca pos Marc Pichel