W. J. Parak
Inorganic Nanoparticles -From Synthesis to Biological
Applications
Organo-metallic synthesis of
inorganic nanoparticles + phase
transfer to aqueous solution
- synthesis
Bioconjugation
- properties
- model-system DNA-Au
- hybrid materials
- particles with single molecules
- phase transfer
- particles saturated with molecules
- introduction of functional groups
- transfer to other particles / molecules
- polymer capsules
(biological) Applications
- Light-addressable electrodes
- Phagokinetic tracks (motility assay)
- Drug delivery
- Smart Sensors
1
CdSe magic-size nanocrystals
S. Kudera, M. Zanella, C. Giannini, A. Rizzo, Y. Li, G. Gigli, R. Cingolani, G. Ciccarella, W. Spahl, W. J. Parak, L. Manna,
2
"Sequential growth of magic-size CdSe nanocrystals", Advanced Materials 19, 548-552 (2007).
Hybrid materials: two functionalities within one particle
(adopted from the
synthesis protocol of H.
Gu et al., JACS, 2004)
FePt
FePt@CdS
one material grown on top of another ? potentially two functionalities within of particle (e.g. fluorescent + magnetic)
many material combinations are possible: FePt@(CdS, ZnS, PbS, CdSe, ZnSe, PbSe) ? very general synthesis route
but: are properties of the original materials retained?
? to our experience: slightly reduced magnetic moment + strongly quenched fluorescence
problems with this concept for hybrid particles: hydrophobic, decreased performance of functionality ? surface modification
M. Zanella, A. Falqui, S. Kudera, L. Manna, W. Parak, in preparation
3
Transfer to aqueous solution: Polymer coating procedure
wrap particles in an amphiphilic polymer shell
works for all particles with hydrophobic stabilizer chains
T. Pellegrino, L. Manna, S. Kudera, D. Koktysh, A. Rogach, S. Keller, J.Rädler, G. Natile, W. J. Parak, Nanoletters 4, 703-707 (2004)
4
Synthesis of the amphiphilic polymer
O
O
O
O
O
O
O
=
O
O
O
O
O
O
O
O
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NH2
HN
HO
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HO
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HN
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HN
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O
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HO
O
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O
HN
O
HO
O
HO
O
O
O
=
HN
=
one monomer
5
Direct embedding of functional groups in the polymer shell
functional group:
- PEG
- organic fluorophore
- sugar
- biotin
- ....
we can directly incorporate functional groups into
the polymer shell without the need for post
bioconjugated chemistry
Cheng-An J. Lina, Ralph A. Sperling, Jimmy K. Li, Ting-Ya Yang, Pei-Yun Li, Marco Zanella, Walter H. Chang, Wolfgang J. Parak ,
submitted to SMALL
6
Direct embedding of functional groups in the polymer shell
O
O O
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O O O
O O
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organic solvent
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aqueous
solution
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n
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Cheng-An J. Lina, Ralph A. Sperling, Jimmy K. Li, Ting-Ya Yang, Pei-Yun Li, Marco Zanella, Walter H. Chang, Wolfgang J. Parak ,
submitted to SMALL
7
General bioconjugation (with controlled number of single molecules)
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NH
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NH
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N
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NH
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O
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HN
H
N
-O
O
O
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O
O
OH
O
O
O
O
O
O
O
-
O
- O
O
O
NH
O
O
O
O
O
O
O
O
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O
-
O
O
O
O
O
O
O
H
O
HO
O
O
Au
Fe3O4
CdSe/ZnS
R. A. Sperling, T. Pellegrino, J. K. Li, W. H. Chang, W. J. Parak, Advanced Functional Materials, 16, 943-948 (2006).
Cheng-An J. Lina, Ralph A. Sperling, Jimmy K. Li, Ting-Ya Yang, Pei-Yun Li, Marco Zanella, Walter H. Chang, Wolfgang J. Parak ,
submitted to SMALL
8
Commonly used quantum-dot based FRET assemblies
qdot excitation
radiation-less
energy transfer
(FRET)
acceptor
(organic dye)
emission
donor (qdot)
emission
competitive
displacement of dyelabeled analyte
with analyte
ligand exchange +
linkage of recognition site
(occupied by organic fluorophore)
-: low colloidal stability
+: close acceptor / donor distance
hydrophobically
capped quantum
dot
polymer-coating / PEGylation +
linkage of recognition site
(occupied by organic fluorophore)
+: high colloidal stability
-: big acceptor / donor distance
9
Polymer with embedded organic fluorophores
Sample purification with gel electrophoresis
+
1
2
3
S F
Polymer coating yields next to polymer-coated particles also some empty micells!
María Theresa Fernández-Argüelles, Aleksey Yakovlev, Camilla Luccardini, Ralph Sperling, Stéphane Gaillard, James Lin, Alfredo Sanz Medel,
10
Walther Chang, Jean-Maurice Mallet, Jean-Marie Brochon, Anne Feltz, Martin Oheim, Wolfgang Parak, Nanoletters, in press.
FRET of acceptor embedded in polymer around donor
fast bands
+
-
1xPA
1xPQD
1xPAQD
4xPAQD
8xPAQD
1
2
3
4
5
S F
7
6
5
4
I(? ) 3
2
1
0
500
600
700
800
? [nm]
slow bands
100000
sample
?
[ns]
1xPQD
10
1xPAQD
2.5
4xPAQD
2.1
8xPAQD
1.7
80000
60000
40000
20000
0
0
10
20
30
7
6
5
4
I(? )
3
2
1
0
500
600
700
800
? [nm]
María Theresa Fernández-Argüelles, Aleksey Yakovlev, Camilla Luccardini, Ralph Sperling, Stéphane Gaillard, James Lin, Alfredo Sanz Medel,
11
Walther Chang, Jean-Maurice Mallet, Jean-Marie Brochon, Anne Feltz, Martin Oheim, Wolfgang Parak, NanoLetters, in press.
Polymer capsules as multifunctional containers
form polyelectrolyte onion-shells (PSS/PAH) around template + dissolve template
load with fluorescent (+ magnetic + metallic) nanocrystals + biological molecules
= loading of a "container" with particles of different functionality
positively
charged
(see work of G. Decher, H. Möhwald)
negatively
charged
functional nanoparticles can be embedded
into wall of capsules by electrostatic attraction
? functionality
size of capsule: ? m
size of nanoparticles: nm
load can be embedded into interior of capsules by
- impregnation of the template core
-swelling + shrinking of the mesh of the walls at different pH
? transport container
capsule preparation by group of G. Sukhorukov
12
Targeted local delivery and release
Capsule uptake by living cells
„hard“ capsules
(template core not removed)
walls labeled with dye
„soft“ capsules
(template core removed)
walls labeled with dye
"hard" and "soft" capsules are ingested by the cells; the "soft" capsules are deformed
MCF-7 cells
confocal microscopy: Maximilian Semmling
A. Muñoz_Javier, S. Kempter, O. Kreft, M. Semmling, A. Skirtach, O. Bruns, J. Rädler, J. Käs, G. Sukhorukov, C. Plank, W. Parak, in preparation
13
Targeted local delivery and release - Magnetic direction of capsules
A
B
C
Magnet
distance AC: 11 mm
distance BC: 5 mm
A
B
C
B. Zebli, A. S. Susha, G. B. Sukhorukov, A. L. Rogach, W. J. Parak, Langmuir 21, 4262-4265 (2005).
14
Targeted local delivery and release
Capsule opening with a light pointer
A. Skirtach, A. Munoz Javier, O. Kreft, K. Köhler, A. Piera Alberola, H. Möhwald,
W. J. Parak , G. Sukhorukov, Angewandte Chemie, 2006
15
Capsules as intracellular sensors: pH-sensitive capsules
embed sensitive dyes (e.g. pH sensitive, Ca sensitive) in polymer capsules ?
- dye is protected against degradation, cell is protected from free dye ? long term measurements
- many dye molecules are embedded in one capsule ? sensitivity
- dye is confined to capsules ? localization
Emissionspectra (Fluorescence) of SNARF-Dextrane in Solution (pH 2 to 11)
acidic pH:
green fluorescence
fluorescence intensity (a.u.)
12000
SNARF: pH sensitive dye
pH 2.0
pH 3.0
10000
8000
6000
alkaline pH:
red fluorescence
capsule
preparation by
group of G.
Sukhorukov
pH 4.0
pH 5.0
pH 6.0
pH 7.0
pH 8.0
pH 9.0
pH 10.0
pH 11.0
4000
2000
0
510 520 530 540 550 560 570 580 590 600 610 620 630 640 650 660 670 680 690 700 710 720 730 740 750
wavelength (nm)
O. Kreft, A. Munoz Javier, G. Sukhorukov, W. J. Parak, Journal of Materials Chemistry, in press
nm
16
Capsules as intracellular sensors: pH-sensitive capsules
acidic pH:
green fluorescence
alkaline pH:
red fluorescence
0 min
30 min
capsules in
extracellular medium:
alkaline environment
capsules ingested by
cells in endosomes:
acidic environment
60 min
180 min
O. Kreft, A. Munoz Javier, G. Sukhorukov, W. J. Parak, Journal of Materials Chemistry, in press
17
Capsules as intracellular sensors: multiplexed measurements?
capsules can carry fluorescence molecules at two disctinct positions:
cavity ? ion sensitive dye
wall ? spectral barcode ? quantum dots
problem: release of capsules for endosome to cytoplasm
18
Capsules as intracellular sensors: multiplexed measurements?
phase
contrast
wall label
TRITCAlexa488channel
channel
greenchannel
SNARF
redchannel
overlay
pH = 6
pH = 9
pH = 6
pH = 9
O. Kreft, A. Munoz Javier, G. Sukhorukov, W. J. Parak, Journal of Materials Chemistry, in press
19
Acknowledgement
Ralf Sperling
Stefan Kudera
Tim Liedl
Almudena Muñoz Javier
Bernd Zebli
Marco Zanella
Maite Fernandez Argüelles
Christian Kirchner
Maximilian Semmling
Feng Zhang
Pablo del Pino
Hermann Gaub
Teresa Pellegrino
Liberato Manna
A. Paul Alivisatos
Carolyn A. Larabell
Josef Käs
Aleksey Yakovlev
Camilla Luccardini
Stéphane Gaillard
Jean-Maurice Mallet
Jean-Claude Brochon
Anne Feltz
Martin Oheim
Eric Dulkeith
Moritz Ringler
Thomas Klar
Andrei Susha
Andrey Rogach
Jochen Feldmann
Alicia Piera
Susi Kempter
Joachim Rädler
Jimmy Li
James Lin
Walter Chang
Alfredo Sanz Medel
Christian Stoll
Fred Lisdat
Oliver Kreft
Andrei Skirtach
Mathieu Bedard
Gleb Sukhorukov
Oliver Bruns
Birgit Bußmann
Tobias Schrader
Stephan Malkmus
Markus Braun
20