Nanotechnology for Molecular and Cellular Manipulation
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1 Nanotechnology for Molecular and Cellular Manipulation Logan Liu Micro and Nano Technology Lab Department of Electrical & Computer Engineering University of Illinois
2 Physical Systems Nano vs. Bio Micro / Nano Life Systems Courtesy of Office of Basic Energy Sciences Office of Science, U.S. DOE Self-assembled, Nature-inspired structure Many 10s of nm Pollen grain Red blood cells Zone plate x-ray lens Outer ring spacing ~35 nm Nanotube electrode MicroElectroMechanical (MEMS) devices m wide Nanoworld Microworld 10-3 m 10-4 m 10-5 m 10-6 m 10-7 m 10-8 m 1,000,000 nanometers = 1 millimeter (mm) Ultraviolet Visible Infrared Microwave 0.1 mm 100 m 0.01 mm 10 m 1,000 nanometers = 1 micrometer ( m) 0.1 m 100 nm 0.01 m 10 nm Dust mite 200 m Human hair ~ m wide Red blood cells (~7-8 m) ~10 nm diameter Fly ash ~ m ATP synthase Quantum corral of 48 iron atoms on copper surface positioned one at a time with an STM tip Corral diameter 14 nm Carbon buckyball ~1 nm diameter Carbon nanotube ~1.3 nm diameter 10-9 m m Soft x-ray 1 nanometer (nm) 0.1 nm DNA ~2-1/2 nm diameter Atoms of silicon spacing nm
3 Outline Molecular Manipulation Movement Conformation Change Cellular Manipulation Movement Growth Gene Expression
4 Moving and Collecting Molecules Nanoscale Molecular Dielectrophoresis Magnetic Nanoparticle Molecular Capture Nano-optical &Nano photonics Molecular Trapping Nanophotonic/Phononic Thermophoretic Molecular Concentration
5 Dielectrophoresis > Clausius-Mosotti Factor Credit : R. Pethig, Bangor Univ.
6 Nanoscale Dielectrophoresis Zheng et al. IEEE Nano (2003)
7 Magnetic Nanoparticle Capturing Invitrogen Dynabeads Nam et al, Science (2003)
8 Optical Trapping
9 Nanoscale Optical Field Gradient Electrostatic Approximation (b) Finite Element Simulation 16 4dB (, ) Solve Laplace equation 1 ( ) E 1 2 Charge density 2 2 a 0 ( / ) 1,0 Singularity near the sharp tip ( 0) y z x Solve Harmonic Wave Equation ( ( E) E 2 H ) H For Plane Wave E k z ( H z ) k 2 0 Ez H z 0 Low-reflecting Boundary Condition n H E z 2 E0z n E H z 2 H 0z 0 0 E / E 0 at 785 nm 9 db 30 db db
10 Optical Trapping on Nanoplasmonic Particle Calander et al PRL (2002)
11 Nanoscale Optofluidic Trapping Yang et al, Nature (2009)
12 Thermophoretic Molecular Trapping Braun et al PRL (2002)
13 Nanoparticle Assisted Thermophoretic Manipulation Light Ilumination Microfluidic Channel Photothermal Nanocarpet Optical Pre-Concentration of DNA Biomolecule concentrating Credit : Logan Liu
14 Changing Molecular Conformations Mechanical Force Electric Force Thermodynamic Control Photochemical Control
15 Stretch Molecules Smith et al, Science (1996) Wang et al, Biophys. J. (1997)
16 Electric Force Unravel Molecules in Nanotubes Kaji et al Biophys. J. (2002)
17 Thermodynamics Controls Molecular Conformation ssdna1 + ssdna2 dsdna Free Energy Association Constant K G 0 RT ln K [ dsdna ] 2 [ ssdna ] [ dsdna ] [ C] 1 [ ssdna ] 2 Increase Temperature T At equilibrium G 0 0 T K [ dsdna] [ ssdna] Free Energy G 0 H 0 T S0 H 0 T S0 RT ln k [ dsdna] [ ssdna] 1 2 At Melting Temperature T m T m H 0 Tm S 0 RTm ln(4 /[ C]) S0 R ln(4 /[ C]) H 0
18 Optically-Controlled DNA Release Steady State 57 ο C 52 ο C 47 ο C 42 ο C 37 ο C 32 ο C 27 ο C Temperature ( C) Steady state 0 ns 1 ns 3 ns 32 ns 50 nm Distance (nm) Lee et al, Nano Letters (2009)
19 Optical Melting dsdna Normalized Absorbance aspect ratio 3.2 aspect ratio 2.5 aspect ratio Wavelength (nm) Normalized Fluorescence Energy (mj/cm 2 ) 50ºC 63ºC 70ºC 15 bp 25 bp 50 bp Time (s) Time (s) Normalized Fluorescence nm 685 nm Time (min) Normalized Fluorescence mw/cm 2 19 mw/cm 2 46 mw/cm Time (min) Lee et al, Nano Letters (2009)
20 RF Magnetic Control of DNA Conformation Credit : Joe Jacobson, MIT Hamad-Schifferli et al. Nature (2002)
21 Photochemical Control of Molecular Conformation Fortin et al, Nature Methods (2008)
22 Moving and Collecting Cells Magnetic Nanoparticle Cell Capture MEMS and NEMS Cell Manipulator Holographic Optical Trapping Optoelectronic Cell Tweezer
23 Moving Cells Using Magnetic Nanoparticles Gu et al. JACS (2003)
24 MEMS/NEMS Cell Manipulation Chronis et al, JMEMS (2005)
25 Holographic Optical Trapping Array Credit : David Grier, NYU
26 Optoelectronic Tweezer Chiou et al, MEMS (2004)
27 OET Cell Manipulation Chiou et al, Nature (2005)
28 Control Cell Growth and Differentiation Surface Nanopatterns 3D Nano scaffold Micro/Nanofluidics Nanoelectrodes
29 Protein Nanopatterning Changes Cell Signaling Mossman et al, Science (2005)
30 Cell Culture in Nanofiber Scaffold Patel et al, Nano Letters (2007)
31 Micro/Nano Fluidic Control of Stem Cell Development Lucchetta et al, Nature (2005)
32 Electrical Neuron Stimulation on Carbon Nanofiber Electrodes Nguyen et al, IEEE Trans. Biomed. Eng. (2007)
33 Control Genetic Profiles Nanophotonic Activation of antisense DNA release in living cells Nanophotonic Activation of sirna gene silencing in living cells
34 Nanophotonics Controlled Genetic Interference Translation Optical Remote Control mrna Protein Antisense Gene Therapy
35 Localized Optical Activation of Gene Release In Vitro HER-2 Antisense DNA Delivery 3 - CTCCATGGTGCTCAC-5 In Vivo Lee et al, Nano Letters (2009)
36 HER-2 Inhibition by Gene Release a b Unbound antibody 1 min Illumination antibody HER-2 untreated cell fluorescent dye treated cell ONCOS blocks protein translation Laser 10 m 10 m BT474 Cell Culture
37 Nanophotonics Controlled SiRNA Gene Silencing Braun et al, Nano Letters (2009)
38 The End Thanks! Questions?
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