Outline - CMU€¦ · Hydrogel swells causing red-shift washing-enzyme titrant creatinine diffuses...
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Nanotechnology Nanotechnology-based Materials and Devices for based Materials and Devices for Biomedicine Biomedicine Hong Koo Kim Co-Director [email protected] Institute of NanoScience and Engineering University of Pittsburgh Outline Outline n Single Single-molecule Studies of Chromatin molecule Studies of Chromatin Fibers Fibers n Bacteriophages Bacteriophages: Molecular : Molecular Nano Nano-machines machines n Crystalline Colloidal Arrays: Molecular Crystalline Colloidal Arrays: Molecular Sensing Sensing Photonic Photonic Crystals Crystals n Nanophotonic Nanophotonic Chip Technologies for Chip Technologies for Instrumentation Instrumentation
Transcript of Outline - CMU€¦ · Hydrogel swells causing red-shift washing-enzyme titrant creatinine diffuses...
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NanotechnologyNanotechnology--based Materials and Devices for based Materials and Devices for BiomedicineBiomedicine
Hong Koo KimCo-Director
Institute of NanoScience and Engineering
University of Pittsburgh
OutlineOutline
nn SingleSingle--molecule Studies of Chromatin molecule Studies of Chromatin FibersFibers
nn BacteriophagesBacteriophages: Molecular : Molecular NanoNano--machinesmachinesnn Crystalline Colloidal Arrays: Molecular Crystalline Colloidal Arrays: Molecular
Sensing Sensing Photonic Photonic CrystalsCrystalsnn Nanophotonic Nanophotonic Chip Technologies for Chip Technologies for
InstrumentationInstrumentation
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Nanoscience Nanoscience and Technologyand Technology
nn 11--100 100 nm length scale: atomic, molecular or nm length scale: atomic, molecular or macromolecular levelsmacromolecular levels
nn Properties not predictable from those at micro or Properties not predictable from those at micro or macro scalemacro scale
nn New phenomena associated with surface/interface New phenomena associated with surface/interface effects and quantum size effects in confined effects and quantum size effects in confined structures structures
nn New modes of transports, collective phenomena, New modes of transports, collective phenomena, chemical chemical reactivitiesreactivities, physical properties, etc., physical properties, etc.
nn In living systems:In living systems:nn multiple length scales and functions (molecules and chemical multiple length scales and functions (molecules and chemical
pathways, subpathways, sub--cellular mechanical structure, cells, organs cellular mechanical structure, cells, organs and tissues)and tissues)
Hierarchical Structure of ChromosomeHierarchical Structure of Chromosome
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SingleSingle--molecule Approach to Chromatin Structure molecule Approach to Chromatin Structure and Dynamicsand Dynamics
Visualization of the three dimensional arrangement of nucleosomes in the chromatin fiber
Atomic Force Microscope After linker histone removal
S.H. Leuba and J. Zlatanova, Eds. (2001) Biology at the single-molecule level, Pergamon
Optical TrappingOptical Trapping
A. Ashkin, PNAS, 94, 4853 (1997)
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Unfolding Individual Unfolding Individual NucleosomesNucleosomesOptical tweezers studyOptical tweezers study
S.H. Leuba et al., Nature Struct . Biol. 8, 606 (2001)
Assembly of Single Chromatin FibersAssembly of Single Chromatin FibersMagnetic tweezers studyMagnetic tweezers study
S.H. Leuba et al., PNAS, 100, 495 (2003)
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PhagesPhagesThe small but powerful living deadThe small but powerful living dead
Small genomics (~104 bp)Evolution over 3.5 billion yearsGlobal population > 1030 phages
R. Hendrix and G. Hatfull
Fighting back: Fighting back: Using viruses to attack bacterial pathogensUsing viruses to attack bacterial pathogens
• Bacterial pathogens: natural killers and potent bioweapons
• Bacteriophages (viruses that infect bacteria) are their natural enemies
• Phages - or phage derivatives - can be employed in many ways to battle pathogens and bioweapons
Phage “Corndog”Anthrax
• Identify organism
• Determine drug sensitivity
• Immediate & effective treatment
Tuberculosis and Anthrax:
Common Strategies
G. Hatfull
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LuciferaseLuciferase Reporter PhagesReporter PhagesRapid, sensitive detection of bacterial cellsRapid, sensitive detection of bacterial cells
No dr
ug
Rifam
picin
Isonia
zid
Strep
tomyci
n
wt
Rif-R
INH-R
Strep-R
Phage-delivery of a reporter gene
Drug-susceptibility testing
700000 70000 7000 700 70 7 > 1 cell > 1 cell
102
103
104
105
106
107
colony forming units
Luci
fera
se a
ctiv
ity (R
LU)
Single cell detection
Simple
Inexpensive
G. Hatfull
S. Asher, Dept of Chemistry
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Dialysis / Ion Exchange Resin
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Crystalline Colloidal Array SelfCrystalline Colloidal Array Self--AssemblyAssembly
Polymerized CCA FabricationPolymerized CCA Fabrication
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Hydrogel swellscausing red-shift
washing
-
enzyme
--titrant
creatinine diffuses into hydrogel and binds to enzyme
creatinine hydrolysisproduces OH -
OH - deprotonatestitrant
OH -
-
= = = creatinine
General Clinical Sensing MotifGeneral Clinical Sensing Motif
Ext
inct
ion
Wavelength / nm
0
0.5
1.0
1.5
2.0
400 450 500 550 600 650
water
41ppb 410 ppb
4.1 ppm
Pb 2+
Pb 2+
Pb 2+
Pb 2+
Pb 2+
Pb 2+
Pb 2+
Pb 2+
Dif
frac
tio
n W
avel
eng
th S
hif
t / n
m
Lead Concentration / ppm
0
20
40
60
80
100
120
140
160
0.02 0.2 2 20 200 2000
CO
O
O
O O
O
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N H
N H
O C
H
N H
O C
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Pb 2 +
H
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Chelation of Pb2+ ion results inimmobilization of the counter ion.Osmotic pressure swells the gel,which results in the red shift of diffraction.
S. Asher et al., Anal Bioanal Chem, 373, 632 (2002)
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PhotonicPhotonic Crystal Glucose SensingCrystal Glucose SensingResponse to glucose in artificial tear fluidResponse to glucose in artificial tear fluid
5 2 5 5 5 0 5 7 5 6 0 0 6 2 5
G l u c o s e c o n c e n t r a t i o n i n m M
1 0 2 0 1 0
Diff
ract
ion
inte
nsity
/a.u
.
W a v e l e n g t h / n m
2 mM tris-HCl, pH 8.5;150 mMNaCl
S. Asher et al., Anal. Chem. 75, 2316 (2003)
PointPoint--ofof--Care Clinical SensorsCare Clinical Sensors
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QuantumDot Corp.
Quantum Dot Quantum Dot FluorophoresFluorophores
d
λ
T/f ~ (d/λ)4 T ?
Transmission of Light through Transmission of Light through SubSub--wavelength Apertures in Metal Filmswavelength Apertures in Metal Films
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Optical Interactions in Metallic Optical Interactions in Metallic Nanoaperture Nanoaperture ArraysArraysSlit width: 80 nmSlit width: 80 nmFDTD simulation
Applications:
Nano-optic filters, beam shaping and
concentrating devices beyond diffractive optics
H.K. Kim et al., Appl. Phys. Lett. 83, 3021 (2003)
NanoNano--optic optic MultispectralMultispectral Fluorescence DetectionFluorescence Detection
multispectral fluorescence signals
monochromator/detectorarray chip
excitation
antibody/protein/antibody-fluorophores
H.K. Kim
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Protein ArraysProtein Arrays
High-density sensor arrays and electrochemical electrodes (nanoprobearrays)
Functional Functional nanoelement nanoelement arrays on a chiparrays on a chip
Template/Molding/Replication
Separation (such as antibody-based nanomembranes for drug separation) and absorbent media, catalytic surface and supports
H.K. Kim et al., Appl. Phys. Lett. 81, 3458 (2002)
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NanosystemsNanosystems--onon--aa--chip based on selfchip based on self--organized organized nanochannelnanochannelarraysarrays
Electronics Photonics
Magnetics/Spintronics
Mechanics/Fluidics
Biochemistry
LogicMemoryCommunicationControlSensingSeparationAbsorbentActuation
Pore diameter: > 5 nmInter-pore distance: > 50 nm(pore density ~10 11 cm-2)
Nanopore arrays formed on silica witha directed self-organization method
Single-domain, square-lattice arrayof holes over entire chip area (~cm2)
