2013 biochip technologies 1 materials in the life sciences
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IMTEK
powerpoint
template
2008:Version2ofthefirstslide
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Content
Materials
and
surface
modifications
(26.04.13)
Manufacturing
of
Biochips
(14.06.13)
Biochiptechnologies Betweenresearchandroutinediagnostics(stateoftheart, 21.06.13)
Nucleicacidbasedtechniques(28.06.13)
Biochips
for
protein
analytics
(05.06.13)
Otherapplications(12.07.13)
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ummary
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Ourprofile
esearc an eac ng
22 faculties
highly interdisciplinary world of
microsystem technology
IMTEK and industry
MSTBw
Core competences of CPI
Preparation of surfaces with tailor-made
Topological and chemical microstructuring of surfaces
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AFM Biochip-technologies
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Biochiptechnologieshttp://portal.uni-freiburg.de/cpi/biochip-group-dr-brandstetter
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Biochips what are they?(1)
devicesthat can contain anywhere from tens to tens of millions of individual
sensor elements or biosensors
The sensors are acked to ether into a acka e t icall the size of a
microscope slide. Because so many sensors can be put into such a small
area, a huge number of distinct tests can be done very rapidly.
Biochips are often made using the same microfabrication technology used
. , ,
electronic (although they can be).
The key premise behind biochips is, that they can do chemistry on a small
scale. Each biosensor can be thought of as a "microreactor, which does
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chemistry designed tosense a specific analyte.
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Biochips what are they?(2)
Biosensors can be made tosense a wide variety of analytes, including DNA,
rotein antibodies and small biolo ical molecules.
Fluorescence is often used to indicate a sensin event. Automated
microscopy systems can be used to "read" the chip, i.e. determine which
sensors are fluorescing
Most biochips are 2D arrays of sensors placed carefully in a grid
. .
,
microdeposition techniquesare used. The sensors are essentially placed one
at a time, or serially, on the chip.
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Biochips what are they?(3)
HPV_3D_Katrin_N_30s_Cy5
substrat
dot
3
13 microarray
http://en.wikipedia.org/wiki/Biochip#History
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Manufacturingof biochips ingeneral(1)
1. Untreated slide
mixed analyte solution
2. Microarray printing
. mmo sa on
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Manufacturingof biochips ingeneral(2)
step1:print polymer mixed with DNA
step 3:
hybridisation
and
photocrosslinking
via UV-irradiation
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Materials and surface modifications
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Biochip materials (1)
Microscope slide of glass Commercial microscope glass slides
Silica (SiO2) + vitreous silica
- -
Limestone (CaCO3) + borosilicate glass-pyrex
Magnesium Carbonate (MgCO3) + aluminosilicate glass
+ borosilicate glass
Detailled information
Frontiers in biochip technology
by Wan-Li Xing, Jing Cheng
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Published by Birkhuser, 2006ISBN 0387255680, 9780387255682
357 pages
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Biochip materials (2)
Microscope slide of plastic Commercial plastic slides
PMMA (polymethymethacrylate) + PMMA
COC (cyclic olefin copolymer) + TOPAS
Polycarbonate + Polycarbonate
Polypropyrene + Polypropyrene
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Lab Chip, 2007, 7, 856 - 862, DOI: 10.1039/b700322f
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Biochip coatings
directly chemically modified surfaces
n s u syn es s on g ass + ac va e g ass y po y-car o m e,
aminosilane, aldehyde
an za e pro es on unmo e g ass + gra coa ng po ymers on s con g ass
Photocrosslinking on unmodified plastic + plastic-based DNA microarrays using
car o m e c em s ry
+ amine-modified PMMA substrates
+ activated polystyrene, polypropyrene,
polycarbonate (PC) S.A. Fodor, R. Rava, X.C. Huang, A.C. Pease, C.P. Holmes and C.L. Adams. Science 251 (1991) 767773. M.J. Moorcroft, W.R. Meuleman, S.G. Latham, T.J. Nicholls, R.D. Egeland and E.M. Southern. NAR, 2005, Vol. 33, e75.
N. Kimura, R. Oda, Y. Inaki and O. Suzuki. Nucleic Acids Research, 2004, Vol. 32, e68.
H.-Y. Wang,R.L. Malek,A.E. Kwitek,A.S. Greene,T.V. Luu,B. Behbahani,B. Frank,J. Quackenbush, N.H. Lee, Genome Biol. 4 (2003), R5.
M. Dufva, S. Petronis, L.B. Jensen, C. Krag and C.B. Christensen. Biotechniques 37 (2004) 286292, 294, 296.
A. Kumar O. Larsson D. Parodi Z. Lian Nucleic Acids Research 2000 Vol. 28 e98.
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M. Schena, D. Shalon, R.W. Davis, P.O. Brown, Science 270 (1995), 467470.
De Paul S. M., Falconnet D., Pasche S., Textor M., Abel A. P., Kauffmann E., Liedtke R. and Ehrat M.. Anal. Chem. 2005, 77, 5831-5838.
Johnson P. A., Gaspar M. A. and Levicky R. J. Am. Chem. Soc.,2004, 126, 9910-9911.
N. Kimura, T. Nagasaka, J. Murakami, H. Sasamoto, M. Murakami, N. Tanaka and N. Matsubara. Nucleic Acids Research, 2005, Vol. 33, e46.
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2DchipsusingSAMs(selfassembledmonolayers)
typical DNA-chip design:
sequence of the probe
polyT(thymine) tailer
adapted from: E. Southern, K. Mir, M. Shchepinov, Nature Gen., 27 (1999) 5
+ reproducibility (why is acceptance of microarrays below expectations in non-research areas?
Weakness:
+ sensitivity
+ surface properties
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Askyscraperapproach
attachment ofo gonuc eot e pro es
3D
polymer brushes
polymer layer approach allows to
improve the sensitivity
adjust properties of the surface
(hydrophilicity, reactivity)
3D
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polymer networks
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Functional polymermonolayers
chemisor tion of ol mers growth of polymerson surfaces
blockco ol mers
grafting of polymers on
plasma modified
surfaces
via macroinitators
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photochemical attachmentof polymers surface-attachedpolymer networks grafting in between
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Photochemistry of benzophenone
triplet formation upon n,* excitation
biradical reacts with C,H bonds
C O C O C
CCH
350 nmO
H265 nm
hydrogen
abstraction
= 100 s
CC
OHrecombination
Toomey R., Freidank D. and
Rhe J.. Swelling Behavior of
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n, ur ace- ac e
Polymer Networks.Macromolecules, Vol. 37, 2004,
882-887.
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Polymernetworks attached to polymeric substrates
photocrosslinkableovercoat
Me
O OONMesimultaneous crosslinking
Me
polymeric substrate
(e.g. polyurethane)
via pendant benzophenone
units
swelling in
ca. 20 m
~ 1 mm
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Microstructuringinbiochiptechnologies,twoprocedures
I. Contact printing
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http://www.anopoli.com/http://www.anopoli.com/
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Microstructuringinbiochiptechnologies,contactprinting
Omnigrid from GeneMachine
Contact printing procedure
65% humidity, RT
Steel or tungsten needle with reservoir
droplet volume 400 600 pl
droplet diameter 140 200 m
Process variance > 10%
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Microstructuringinbiochiptechnologies,contactprinting
Pin heads make the difference.
Split pinSpot diameters : 75m to 215 m
Uptake volumes : 0.25l to 0.64 l
li.c
om/
http://www.a
nopo
Solid pin
Spot diameters : 75m to 450 m
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Microstructuringinbiochiptechnologies,contactprinting
Printing with different, not aqueous, solutions is possible.
PDMAA(Polydimethylmetacrylate) PS (Polystyrene)
200 m
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printing medium: ethanol printing medium: toluene
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Microstructuringinbiochiptechnologies,contactprinting
Spot diameter is not really controllable.
Split pin
Solid pin
Printing of 0.25 m Cy5-labelled oligo-DNA in 400
mM Napi and 1mg/ml PDMAA-co-5%MABP-co-
2,5%VPA
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Microstructuringinbiochiptechnologies,contactprinting
sca e n ng
PDMAA layer
PMMA 5 m /ml linin
Printing medium toluene
exposure after
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Microstructuringinbiochiptechnologies,contactprinting
1. copolymers 2. buffer 3. PT-6000 tungsten
PDMAA-co-5%MABP-co-2 5%VPA a 400 mM Na i
plastic/PMMA glass/Epoxy
(b) 200 mM Napi/3xSSC/0.75 M betaine
2D
. . ..
2D_16_04_07_P2Dsp.2a 2D_16_04_07_N2Dsp.4b
a. a. b.
2D_04_04_07_N2Ds.4a2D_04_04_07_P2Ds.1a
b.
3D3D
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3D_12_04_07_P3Dsp.11a 3D_12_04_07_N3Dsp.2a 3D_03_04_07_N3Ds.4a3D_03_04_07_P3Ds.11
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Microstructuringinbiochiptechnologies,contactprinting
1. copolymers 2. buffer 3. PT-6000 tungsten
PDMAA-co-5%MABP-co-2,5%VPA (a) 400 mM Napi
plastic/PMMA glass/Epoxy
(b) 200 mM Napi/3xSSC/0,75 M betaine
2D
. . ..
2D_16_04_07_P2Dsp.2a
a. a. b.
2D_xx_04_07_N2Ds.x2D_xx_04_07_P2Ds.x
b.
2D_16_04_07_N2Dsp.4b
3D3D
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3D_12_04_07_P3Dsp.11a 3D_12_04_07_N3Dsp.2a 3D_xx_04_07_N3Ds.x3D_xx_04_07_P3Ds.x
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Microstructuringinbiochiptechnologies,contactlessprinting
II. Contactless printing/Piezo Electric Dispenser
http://www.scienion.de
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Microstructuringinbiochiptechnologies,contactlessprinting
II. Piezo Electric dispenser
Piezo Electric dispenser(Scienion AG)
Contactless printing procedure
um y,
droplet volume 410 pl,
droplet volume and diameter is
adjustable
Process variance < 10%
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Microstr ct ring in biochip technologies contactless printing
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Microstructuringinbiochiptechnologies,contactlessprinting
Photos after print
2D 3D
3D2D = printing with PBS without polymer
3D = printing with PBS 1 mg/ml PDMAA-co-
5%MABP-co-2,5%VPA
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i i i bi hi h l i l i i
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Microstructuringinbiochiptechnologies,contactlessprinting
rop e s ac ng
1mg/ml polymer in distilled water
=
PMMA = Polymethylmetacrylate
Small droplet with 10x
Large droplets with 20x
Photo after print
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PSS PMMA
Mi t t i i bi hi t h l i t tl i ti
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Microstructuringinbiochiptechnologies,contactlessprinting
onu -s ruc ur ng
1mg/ml PDMAA-co-
5%MABP-co-2,5%VPA inPBS
Exposure after wash with
PBS and 0.1% (v/v) Tween)
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Dot morphology how to analyze?
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Dotmorphology,howtoanalyze?
Dot morphology, depending on
surface properties
print solution contact angle
analyte concentration
,
AFM
Raster electron microscope
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Microstructuring in biochip technologies contactless printing
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Microstructuringinbiochiptechnologies,contactlessprinting
r n ng w a ves, avo ng
donut-morphology
1mg/ml PDMAA-co-5%MABP-co-2,5%VPA in PBS
Additive Glycerol
0 2.5 5 10 25%(v/v)
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Microstructuring in biochip technologies contactless printing
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Microstructuringinbiochiptechnologies,contactlessprinting
r n ng w w ou re a ose
1mg/ml PDMAA-co-5%MABP
-co-2,5%VPA in PBS
-T+T
125 mg/ml Trehalose (T) in PBS
Donut-structure without
Homogeneity in the dot
morphology, using Trehalose
-T
- - - -
http://en.wikipedia.org/wiki/Trehalose
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glucopyranoside(,
Trehalose) Exposure with a fluorescence microscope
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MicrostructuringMicrostructuring ininbiochipbiochip technologiestechnologies
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MicronasMicronas BiochipBiochip technlogytechnlogy
Piezo Electric dispenser(Scienion AG)
Contactless printing procedure
80% humidity, RT
droplet volume 390 pl,
photodiode diameter 180 m
pr n ng on s ruc ure sur aces
Process variance < 10%
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Microstructuringinbiochiptechnologies
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MicronasBiochiptechnlogy
Piezo Electric dispenser(Scienion AG)
printing directly on a photodiode
180 m
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Microstructuringinbiochiptechnologies
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MicronasBiochiptechnlogy
Piezo Electric dispenser(Scienion AG)
printing directly on a photodiode
pattern matching using a software
ed
notprin
d
printe
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MicrostructuringMicrostructuring inbiochiptechnologies,summaryinbiochiptechnologies,summary
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gg p g , yp g , y
Piezo Electric dispenser (Scienion AG)
Droplet volume control
Droplet diameter tunable (>100m)
r nt ng on y w t aqueous so ut ons
1mg/ml polymer
Process variance < 10%
Omnigrid from GeneMachine
Contact rintin rocedure
Steel or tungsten needle with reservoir
droplet volume 400 600 pl
.
Printing of different solutions
> 1mg/ml polymer possible
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Process variance > 10%
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Thank you for your attention!
http://www.bilder-welten.net/de/produkt_detail.php?id=23019&catid=1623
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Literature
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E. Southern, K. Mir, M. Shchepinov, Nature Gen., 27 (1999) 5
Frontiers in biochip technology, by Wan-Li Xing, Jing Cheng, Edition: illustrated, published by
Birkhuser,2006, ISBN 0387255680, 9780387255682, 357 pages
Lab Chip,2007, 7, 856 - 862, DOI: 10.1039/b700322f
S.A. Fodor, R. Rava, X.C. Huang, A.C. Pease, C.P. Holmes and C.L. Adams. Science 251
(1991) 767773.
. . , . . , . . , . . , . . . . .
NAR,2005, Vol. 33, e75.
N. Kimura, R. Oda, Y. Inaki and O. Suzuki. Nucleic Acids Research,2004, Vol. 32, e68.
H.-Y. Wan R.L. Malek A.E. Kwitek A.S. Greene T.V. Luu B. Behbahani B. Frank J.
Quackenbush, N.H. Lee, Genome Biol. 4 (2003), R5.
M. Dufva, S. Petronis, L.B. Jensen, C. Krag and C.B. Christensen. Biotechniques 37 (2004)
286292, 294, 296.
A. Kumar, O. Larsson, D. Parodi, Z. Liang, Nucleic Acids Research,2000, Vol. 28, e98.
M. Schena, D. Shalon, R.W. Davis, P.O. Brown, Science 270 (1995), 467470.
De Paul S. M., Falconnet D., Pasche S., Textor M., Abel A. P., Kauffmann E., Liedtke R. and
.. . . , , - .
Johnson P. A., Gaspar M. A. and Levicky R. J. Am. Chem. Soc.,2004, 126, 9910-9911.
N. Kimura, T. Nagasaka, J. Murakami, H. Sasamoto, M. Murakami, N. Tanaka and N.
Matsubara. Nucleic Acids Research 2005 Vol. 33 e46.
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Toomey R., Freidank D. and Rhe J.. Swelling Behavior of Thin, Surface-Attached PolymerNetworks. Macromolecules, Vol. 37,2004, 882-887.