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Carbon nanotubesfor
experimental biology
Presented by:
Abdennour ABBAS
Institute of Electronic, Microelectronic and Nanotechnology
University of Lille1
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Table of contents
IntroductionI- What are carbon nanotubes ?
II- Fabrication techniques
III- General applications
IV- Interest in biology
V- Perspectives
Conclusion
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Introduction
Living organisms are built of cells that are typically 10m across. Even smaller are the proteins with a typical
size of just 5 nm, which is comparable with thedimensions of smallest manmade nanomateriel. Thissimple size comparison gives an idea of using
nanomateriel especially Carbon NanoTubesas very smallprobes.
In fact, since carbon nanotubes has been discovered,there is an elevated interest in performing single-cellexperimentation and manipulation including single-cellsurgery, precise drug release, and spatially resolved cell
function monitoring.
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I- What are carbon nanotubes?
Carbon nanotubes (CNTs), long, thin
cylinders of carbon, were discovered in1991 by S. Iijima.
Nanotubes are formed by rolling up agraphene (one-atom-thick layer of graphite)sheet into a cylinder and capping eachend with half of a fullerene molecule(simillar to half C60).
S. Iijima.
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CNTs properties
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Structural properties
diameter = nanometers
length = centimeters
TEM image of nanotube filled with samarium oxide
There are two main types of nanotubes:
single-walled nanotubes (SWNTs) and
multi-walled nanotubes (MWNTs).
A nanotube (also known as abuckytube) is a member of the fullerene
structural family,
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Single-walled nanotubes
(SWNTs):
Diameter = 1nm Tube length = 10mic.m
=> aspect ratios (length/diameter) ofover ten million, which is extremelyadvantageous.
They exhibit important electric
properties that are not shared by themulti-walled carbon nanotube.
SWNTs are the best candidate for
miniaturizing electronics past themicroelectromechanical scale.
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Multi-walled nanotubes (MWNTs):
MWNT possess severalgraphitic concentric layers.
Distance between each layer
is 0.34 nm, with diameters from10 to 200 nm and lengths up tohundreds of microns
MWNT are mainlymonodispersed
Electron microscopy of raw MWNTsimaged by: (a) SEM; and (b)TEM.(R.P. Raffaelle et al. / Materials
Science and Engineering B 116(2005) 233243)
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Assembled CNTs:
Until recently, assembly of individualnanotubes reproducibly into submicron
diameter bundles was not feasible.
The dense bundling (in SWNTs) is
attributed to a strong van der Waalsinteraction.
the assembly process is irreversible.D: microscope imageconfirming excellent stabilityof CNT-probe uponimmersion in a water droplet.(Kouklin et al. Appl. Phys.Lett. 87, 173901 2005).
CNTs bundle
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Schematics of the assembly mechanismby modified dielectrophoretique basedtechnique (Kouklin et al. Appl. Phys. Lett.
87, 173901 2005).
SEM images of MWNT-bundle left coated withsilicone.
The inset shows a detailed view of a typicalMWNT bundle-electrode contact area with no
coating applied, bar is 1 m.
Assembly technique:
Ethanol solution (90%)
Electrode(electric fieldof 105 V/m)
Droplets surface tension
CNTs Probe
CNTs
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very good thermal conductors along the tube "ballisticconduction,
Electrical properties
SWNT ===>
MWNT ===> only semi-conducting
Large Youngs modulus (=1,1 million MPa , Fe=
196 000) High strength
non-biodegradable.
Physical properties(Ajayan et al. PNAS u December 7, 1999 u vol. 96 u no. 25 u 1419914200)
semi-conducting (Zigzag (7,0) nanotube )
metallic (Armchair (5,5) nanotube )
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Chemical properties
Nanotubes are composed entirely of sp bonds, similarto graphite.
Nanotubes naturally align themselves into "ropes" heldtogether by Van der Waals forces.
Under high pressure, nanotubes can merge together.
Nonpolar properties:this nonpolarity enables thecoupling of membrane-constituent lipids to the surface ofthe CNT probes.
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II- Synthesis techniques
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Fullerenes and carbon nanotubesare commonly formed in such mundaneplaces as candle flames. However,
these naturally occurring varieties are:
highly irregular in size and quality,
the high degree of uniformitynecessary to meet the needs of
research,
industry is impossible in such an
uncontrolled environment.
Naturally synthetisied CNTs
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CNTs synthesis techniques:
High-Pressure CO Conversion (HiPCO) Pulsed-Laser Vaporisation (PLV) Carbon Arc Discharge (CA)
Chemical Vapor Deposition (CVD)
The CVD method has shown themost promise in being able toproduce larger quantities ofnanotube (compared to the othermethods) at lower cost. This isusually done by reacting a carbon-containing gas (such as acetylene,ethylene, ethanol, etc.) with a metal
catalyst particle (usually cobalt,nickel, iron) at temperatures above600C.
Nanotube production by CVD(J.H. Hafner et al. / Progress in Biophysics &Molecular Biology 77 (2001) 73110)
catalyst nanoparticles
Hydrocarbon carbon
heat
adsorbtion and precipitation
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III- General applications
For nanoelectromechanical systems(NEMS).
CNT offers possibilities to create futurenanoelectronics devices: transistors, circuits,and computers.
Great interest in quantum physics.
Use of nanotubes in incandescent lamps, replacing atungsten filament (Louisiana State University 2004)
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IV- Interest in biology
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===> before any further use for biology, CNTs mustundergo:
purification (chromatography, centrifugation, filtration)
Solubilization (covalent or noncovalent functionalizationwith surfactant: SDS, ..)
===> BiocompatibilityCytotoxic (C. Klumpp et al. / Biochimica et Biophysica Acta, (2005) -
article in press-)
good biocompatibility (J. Chopek et al. / Carbon - article in press-(2006))
That depends on : purity, composition of CNTs, kind ofcell.
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Typical configurations utilised in nano-biomaterials applied to medical or biological
problems (OV Salata, Journal of Nanobiotechnology2004, 2).
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Overview of possible reactions for the functionalisation of the carbon nanotubesidewall.
(Z.P. Xu et al. / Chemical Engineering Science 61 (2006) 1027 1040)
Functionalisation of the carbon nanotube
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Free of permeabilisation and precise delivery:
Permeabilisation protocols Undesirable side effects (apoptosis,
necrosis)Dont diffuse in sufficient
quantity CNTprobe penetrate easily the membrane of epithelial cell andcontact the nuclear membrane.
Fluorescent left andphase-contrast imagesof the cell afterpenetration with aCNT probe onto the tipof which Alexa 1363
fluoresceinyl glycineamide was adsorbed
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Nanovectors for drug delivery:
The ability of f-CNT to penetrate into the cells offers the potential of using f-CNT as vehicles for the delivery of small drug molecules
(Alberto Bianco et al.,Current Opinion in Chemical Biology 2005, 9:674679)
A- Covalent attachmentof amphotericin B andfluorescein
isothiocyanate to CNT.
B- CNT labelled with a
fluorescent agent were easilyinternalised and could betracked into the cytoplasm orthe nucleus usingepifluorescence and confocal
microscopy
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Carbon nanotubes as electronic devices for sensing inaqueous solutions. An AFM image of a portion of thenanotube network (0.5 m on a side) is shown. (Chen et al.,PNAS April 29, 2003 vol. 100 no. 9)
Sensing in aqueous solutions
100 nm
Carbon
fiber
Biosensor for amperometricmeasurement (M. Yang et al. /Biomaterials 27 (2006) 246255)
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(A) Real-time electronic sensing of specific biological recognitionon nanotubes. Scheme for SA recognition with a nanotubecoated with biotinylated Tween.
Protein and pathogen detection
Specific detection ofmAbs binding to arecombinant humanautoantigen with a
nanotube device coatedwith a U1A antigenTween conjugate.
(Chen et al., PNAS April 29, 2003 vol. 100 no. 9)
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Structural imaging with carbon nanotube AFM probes(Adam T Woolley et al, Chemistry & Biology 2000, Vol 7 No 11)
Schematic of an atomic forcemicroscope.
Isolated IgG molecule imaged with a pore growth MWNT tip(J.H. Hafner et al., 2001 )
Individual GroES molecule detected with asurface growth SWNT tip
Covalent functionalization of carbon nanotube AFM tips:
Surface growth methodforCVD nanotube tip preparation.
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SEMmicrograph shows the nanoscale interactionbetween the neurite and the nanotube bundles. The
arrows show the direction of deformation of thenanotube bundles by the extending neurite.
SEM micrograph indicating guided neurite growthalong long MWNT scaffolds. Neurons show preferentialadhesion as well as proliferation along the MWNTpattern
Scanning electron micrograph demonstratingguided neurite growth along the MWNT array
pattern.
vertical carbon nanotubearrays as support platforms forguiding neurite growth. They can befunctionalized with different biomolecules like neurongrowth factors and adhesion agents ( X. Zhang et al.
/ Sensors and Actuators B 106 (2005) 843850)
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IV- Perspectives
Single-cell experimentation
The attributes, along with their reduced dimensions,make carbon nanotubes a highly attractive platformfor future-generation ultra-sensitive minimally
invasive bio-molecular probing, single-cellexperimentation, and delivery.
Nanoscale imaging and measurementsImproved resolution, sensitivity, and reducedartifact effects in nanoscale imaging andmeasurements.
Best control of CNTs synthesis and purification
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Conclusion
The rediscovery of carbon nanotubes has openednew frontiers in the field of nanotechnology and
nanoscience.
The future experimental biology will rest onmicrodevices and microsystems built fromnanomaterials like carbon nanotubes.
Bibli h
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Bibliography
Ajayan P. M. etal., Carbon nanotubes: From macromolecules
to nanotechnology.
PNAS December 7, 1999 vol. 96 no. 25 u 1419914200
Bianco Alberto et al., Applications of carbon nanotubes in
drug delivery.
Current Opinion in Chemical Biology 2005, 9:674679
Chen Robert J. et al., Noncovalent functionalization of
carbon nanotubes for highly specific electronic biosensors.
PNAS April 29, 2003 ; vol. 100 ; no. 949844989
Chopek J. et al., In vitro studies of carbon nanotubes
biocompatibility.
Carbon (2006) Article in press-
Hafner J.H. et al., Structural andfunctional imaging with
carbon nanotube AFM probes
Progress in Biophysics & Molecular Biology 77 (2001) 73
110
Klumpp Cedric et al ., Functionalized carbon nanotubes as
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experimentation and assays.
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Salata OV etal., Applications of nanoparticles in biology and
medicine.
Journal of Nanobiotechnology 2004, 2
Woolley Adam T et al, Structural biology with carbonnanotube AFM probes.
Chemistry & Biology 2000, 7:R193-R204
Xua Zhi Ping et al., Inorganic nanoparticles as carriers for
efficient cellular delivery.
Chemical Engineering Science 61 (2006) 1027 1040
Zhanga Xuan et al., Guided neurite growth on patterned
carbon nanotubes.
Sensors and Actuators B 106 (2005) 843850
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