There is a plenty of room at the bottom
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1 NanoBiotechnology
2 Lecture by Richard Feynman (1959) There is a plenty of room at the bottom We could arrange the atoms one-by-one the way we want them High-resolution microscopes would allow a direct look at single molecules in biological systems It is very easy to answer many of these fundamental biological questions; you just look at the thing! Unfortunately, the present microscope sees at a scale which is just a bit too crude. Make the microscope one hundred times more powerful, and many problems of biology would be made very much easier.
3 Nanotechnology Creation of useful materials, devices, and systems through the manipulation of matter on nanometer scale. - Generally nanotechnology deals with structures sized between 1 to 100 nanometer in at least one dimension. Ability to design systems with defined structure and function on the nanometer scale. Involves developing materials, devices within that size, and analytical tools (methodology), which can be used for analysis and measurement on a molecular scale Interdisciplinary area : Biology, Physics, Chemistry, Material science, Electronics, Chemical Engineering, Information technology
4 Nanotechnology Plays by Different Rules Normal scale Nanoscale
5 Analytical methods and Nano-sized materials Analytical tools : Atomic force microscopy(afm), Electron microscopy (EM) Nano-sized materials Unusual and different property - Semiconductor nanocrystals: Size-dependent optical property - Nanoparticles: Magnetic nanoparticles (Ferromagnetic, superparamagnetic), Gold nanoparticles, Carbon nanotubes, Graphene - Superparamegnetism: In the absence of an external magnetic field, magnetization is in average zero
6 Graphene Allotrope of carbon in the form of a two-dimensional, atomic-scale, hexagonal lattice Extraordinary properties. - About 200 times stronger than steel by weight - Conducts heat and electricity with great efficiency - Nearly transparent Potential applications: -semiconductor, electronics, battery energy, and composites Andre Geim and Konstantin Novoselov at the Univ Manchester won the Nobel Prize in Physics in Groundbreaking experiments regarding the two-dimensional material graphene
7 Scanning probe microscopy image Graphene-Based Nanomaterials
8 Future implications of nanotechnology Nanotechnology may be able to create many new materials and devices with a vast range of applications, such as in medicine, biomaterials, electronics, and energy production. Nanotechnology raises many of the same issues as with any introduction of new technology, including concerns about the toxicity and environmental impact of nano-sized materials, and their potential effects on global economics.
9 Nano-Biotechnology Integration of nano-sized/structured materials, nano-scale analytical tools, and nano-devices with biological sciences for development of new biomaterials and analytical tool-box as well as for understanding life science Use of bio-inspired molecules or materials Typical characteristics of Biological events/materials - Self assembly - Highly efficient : high energy yield - Very specific : extremely precise Bio-molecules Proteins, DNA, RNA, Aptamers, Peptides, Antibody, Virus
10 Nano-Bio Convergence Bio-inspired device and system Bio-Technology Nano-Technol Molecular Imaging Molecular Switch DNA barcode Biochip / Biosensor Nanotherapy / Delivery Bionano-machine / Nano-Robot
11 Applications and Perspectives of Nanobiotechnology Development of new tools and methods - More sensitive - More specific - Multiplexed - More efficient and economic Implementation - Diagnosis and treatment of diseases - Rapid and sensitive detection (Biomarkers, Imaging) - Targeted delivery of therapeutics - Drug development - Understanding of life science
12 Examples Nano-Biodevices Nano-Biosensors Drug and gene delivery using nanoparticles Imaging with nanoparticles Analysis of a single molecule/ a single cell
13 Issues to be considered Synthesis or selection of nano-sized/ structured materials: bottom-up or top-down Functionalization with biomolecules or for biocompatibility Integration with devices and/or analytical tools Assessment : Reproducibility, Toxicity Mass production and practical implementation
14 The size of things
15 NanoBiotech was initiated by the development of SPM(Scanning Probe Microscopy) that enables imaging at atomic level in 1980 Atomic Force Microscopy (AFM) Scanning Tunneling Microscopy (STM)
16 AFM (Atomic Force Microscope) One of the foremost tools for imaging, measuring, and manipulating matters at the nanoscale. A cantilever with a sharp tip (probe) at its end that is used to scan the specimen surface When the tip is brought into a close proximity of a sample surface, force between the tip and sample leads to a deflection of the cantilever according to Hooke s law (F= -kx) Deflection is measured using a laser spot reflected from the top surface of the cantilever into an array of photodiode
17 Principle and mode of AFM
18 VEECO TESPA VEECO TESPA-HAR NANOWORLD SuperSharpSilicon Tip length : 10 m Radius : 15~20 nm Tip length :10 m (last 2 m 7:1) Radius : 4~10 nm Tip length :10 m Radius : 2 nm
19 Resolution of protein structure by AFM Image of ATP synthase composed of 14 subunits
20 Molecular imaging Biomedical & Biological Sciences : - Ultra-sensitive imaging of biological targets under non-invasive in-vivo conditions - Fluorescence, positron emission tomography, Magnetic resonance imaging Ultra-sensitive imaging - Cancer detection, cell migration, gene expression, localization of proteins, angiogenesis, apotosis - MRI : Powerful imaging tool as a result of non-invasive nature, high spatial resolution and tomographic capability Resolution is highly dependent on the molecular imaging agents Signal enhancement by using contrast agents : iron oxide nanoparticles
21 Semiconductor Nanocrystals Quantum Dots Properties and Biological Applications
22 Synthesis of CdSe/ZnS (Core/Shell) QDs Step 1 CdO + Se CdSe Step 2 Solvent : TOPO, HAD, TOP Surfactant : TDPA, dioctylamine ZnS ZnEt 2 + S(TMS) 2 CdSe Growth temperature 140 (green) 200 (red) CdSe/ZnS 5.5 nm (red) Se solution Ar Thermocouple 320 CdO solution 20 nm Bawendi et al. J. Am. Chem. Soc. (1994)
23 Optical Properties of Quantum Dots a) Multiple colors b) Photostability c) Wide absorption and narrow emission d) High quantum yield Quantum Yield 60 ~ 70 % Single source excitation
24 Coating of QD Surface for Biocompatibility Encapsulation with the hydrophobic core of a micell P O CdSe O P O P O P O P O P O P Coating with PC Coating of the outer Shell with ZnS P O O P ZnS O P CdSe O P O P O P O P NH 2 NH 2 P O O P ZnS O P NH 2 CdSe O P + N O P + N O P O P + N + N + N NH 2 NH 2 NH 2 NH 2 CdSe QDs CdSe/ZnS core-shell Quantum Dots encapsulated in phospholipid micelles NH 2 PEG-PE (n-poly(ethylenglycol)phosphatidylethanolamine): micell-forming hydrophilic polymer-grafted lipids comparable to natural lipoproteins PEG : low immunogenic and antigenic, low non-specific protein binding PC : Phosphatidylcholine Dubertret et al. Science (2002)
25 In vitro imaging Y QD QD-Antibody conjugates + Organelle Antigen Y QD Organelle 3T3 cell nucleus stained with red QDs and microtubules with green QDs - Multiple Color Imaging - Stronger Signals Wu et al. Nature Biotech
26 In vivo imaging Live Cell Imaging Quantum Dot Injection Cell Motility Imaging Red Quantum Dot locating a tumor in a live mouse 10um Green QD filled vesicles move toward to nucleus (yellow arrow) in breast tumor cell Alivisatos et al., Adv. Mater.,
27 Bio-inspired systems Inherent capabilities of molecular recognition and self-assembly Attractive template for constructing and organizing the nano-structures Proteins, toxin, coat proteins of virus etc.
28 α -Hemolysin: Self-assembling transmembrane pore A self-assembling bacterial exo-toxin produced by some pathogens like Staphylococcus aureus as a way to obtain nutrients lysis of red blood cells Alpha-hemolysin monomers bind to the outer membrane of susceptible cells. Monomers oligomerize to form a water-filled heptameric transmembrane channel that facilitates uncontrolled permeation of water, ions, and small organic molecules. Rapid discharge of vital molecules, such as ATP, dissipation of the membrane potential and ionic gradients, and irreversible osmotic swelling leading to the cell wall rupture (lysis), can cause death of the host cell.
29 - Mushroom-like shape with a 50 A beta-barrel stem - Narrowest part (1.4 nm in diameter) of channel at the base of stem
30 Biotechnological applications :Stochastic sensors A molecular adaptor is placed inside its engineered stem, influencing the transmembrane ionic current induced by an applied voltage Reversible binding of analytes to the molecular adaptor transiently reduces the ionic current - Magnitude of the current reduction : type of analyte - Frequency of current reduction intervals: Analyte concentration Stochastic system: systems that are unpredictable due to the influence of a random variable
31 Construction of stochastic sensors
32 a : Histidine captured metal ions (Zn+2, Co+2, mixture ) b: CD captures anions (promethazine, imipramine, mixture) c : biotin ligand
33 DNA sequencing Transmembrane pore can conduct big (tens of kda) linear macromolecules like DNA or RNA Eelectrophoretically-driven translocation of a 58-nucleotide DNA strand through the transmembrane pore of alpha-hemolysin Changes in the ionic current by the chemical structure of individual strands Nucleotide sequence directly from a DNA or RNA strand A single nucleotide resolution
34 DNA sequencing by nanopore
35
36 Understanding Cancer and Related Topics Understanding Nanodevices Developed by: Jennifer Michalowski, M.S. Donna Kerrigan, M.S. Jeanne Kelly Brian Hollen Explains nanotechnology and its potential to improve cancer detection, diagnosis, and treatment. Illustrates several nanotechnology tools in development, including nanopores, quantum dots, and dendrimers. These PowerPoint slides are not locked files. You can mix and match slides from different tutorials as you prepare your own lectures. In the Notes section, you will find explanations of the graphics. The art in this tutorial is copyrighted and may not be reused for commercial gain. Please do not remove the NCI logo or the copyright mark from any slide. These tutorials may be copied only if they are distributed free of charge for educational purposes.
37 What Is NanoBiotechnology? Water molecule Nanodevices Nanopores Dendrimers Nanotubes Quantum dots Nanoshells White blood cell A period Tennis ball
38 Designing Nano-devices for Use in the Body Too Small Too Big
39 Manufacturing Nanodevices Top-down approach: Molding or etching materials into smaller components Bottom-up approach :Assembling structures atom-by-atom or molecule-by-molecule, useful in manufacturing devices used in medicine. X-ray beam Crystal Scattered X-rays Detector Atoms in crystal Crystal Nanodevices White blood cell
40 Nanodevices Are Small Enough to Enter Cells Cell (10,000~ 20,000 nm) Nanodevices Nanodevices Water molecule White blood cell
41 Nano-devices can improve cancer detection and diagnosis at early stages NanoBiotechnology Imaging Physical Exam, Symptoms
42 Nanodevices can improve sensitivity Normal cells Precancerous cells Nanodevices could potentially enter cells Normal cells and determine which cells are cancerous or precancerous. Precancerous cells
43 Nanodevices can preserve patients samples Traditional Tests Cells from patient Cells altered Active state lost Nanotechnology Tests Cells from patient Cells preserved Active state preserved Additional tests
44 Nanodevices can make cancer tests faster and more efficient Patient A Patient B Many diagnostic tests simultanelusly
45 Cantilevers can make cancer tests faster and moreefficient Cancer cell Antibodies with proteins Antibodies Bent cantilever Water molecule White blood cell Nanodevices Cantilevers
46 Nanopores Single-stranded DNA molecule Nanopore A T C G Nanopore A Singlestranded DNA molecule T Nanopore Single-stranded DNA molecule Water molecule Nanodevices Nanopores White blood cell
47 Quantum Dots Ultraviolet light off Ultraviolet light on Quantum dot bead Quantum dots Quantum dots emit light Water molecule Nanodevices Quantum dots White blood cell
48 Quantum dots can find cancer signatures Cancer cells Quantum dot beads Healthy cells Cancer cells Quantum dot beads Healthy cells
49 Improving cancer treatment Traditional Treatment Nanotechnology Treatment Drugs Toxins Nanodevices Cancer cells Cancer cells Toxins Noncancerous cells Noncancerous cells Dead cancer cells Dead cancer cells Dead noncancerous cells Intact noncancerous cells
50 Nanoshells Near-infrared light off Near-infrared light on Nanoshell Gold Nanoshell absorbs heat Water molecule Nanodevices Nanoshells White blood cell
51 Nanoshells as cancer therapy Nanoshells Nanoshells Cancer cells Cancer cells Healthy cells Healthy cells Near-infrared light Dead cancer cells Intact healthy cells
52 Nanodevices as a link between detection, diagnosis, and treatment Traditional Cancer Treatment NanoBiotechnology Cancer Treatment Cancer cell Cancer cell Drug Nanodevice Imaging Reporting Detection Targeting
53 Dendrimers Cancer cell Dendrimer Water molecule Nanodevices Dendrimer White blood cell
54 Dendrimers : Highly Branched Dendritic Macromolecules
55 Poly (amido amine) Dendrimers Characteristics Monodisperse macromolecule Globular (Spherical) Facile surface bio-functionalization Similar molecular size to biomolecules (Glucose oxidase nm) Applications 4.5 nm G4 Poly(amidoamine) Dendrimer Vehicles for delivery of genes and drugs Biomimetic catalysts (Peptides-, Glycodendrimers) Medical applications (MRI contrast enhancer) Molecular carriers for chemical catalysts (Core, Peripheral)
56 Dendrimers as cancer therapy Manipulate dendrimers to release their contents only in the presence of certain trigger (molecules or light) caged molecules Therapeutic agent Cancer detector Reporter Cell death monitor Water molecule Nanodevices Dendrimer White blood cell
57 NanoBiotechnology in Diagnosis and Patient Care Today 2020
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