Gold nanoparticles. and Light. and Bioapplications

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1 Gold nanoparticles and Light and Bioapplications February 2014

2 2 The Incredible Internet...

3 3 ome basics Gold nanoparticles preparation and functionalisation properties Bio- and medical use of Gold nanoparticles

4 4 Light- electromagnetic radiation 1 c E photon h hc

5 5 Quantum mechanical picture of matter ATOM ENERGY DIAGRAM MOLECULE OLID Formation of energy levels in solids Atomic orbitals Molecular orbitals Atom 4 atoms Lots of atoms together =BAND ource: s * 1s s 1s In solid structures (here a metal) the outermost electrons form continuous energy bands from the atom orbitals of the solid. Electron with a particular spin

6 6 Light interaction with matter ABORPTION Absorption PECTRUM of carotene LUMO HOMO Ground state Excited state LUMO=lowest unoccupied molecular orbital HOMO=highest occupied molecular orbital A CHROMOPHORE= part of a molecule that absorbs visible light

7 7 Light interaction with matter EMIION one possible route for relaxation Emission spectrum ource: ource:

8 Nanoparticles- between molecules and bulk 8 Bulk Nanoparticles from ~1 to < 1000 nm Atoms and molecules Energy diagram

9 9 History of colloidal gold... Michael Faraday prepared colloidal gold in his lab Colloidal gold had been used in stained glass from 500 BC. Michael Faraday museum, London The therapeutic use of gold can be traced back to the Chinese in 2500 BC. Colloid: a fluid with particles in the range1 nm-1 mm.

10 Chemical synthesis of gold nanoparticles 10 Precursor solution alt of the desired metal HAuCl 4, AgNO 3 tabilator -> to prevent aggregation (polymers, surface active molecules etc.) Reducing agent -> to reduce the precursor ions into metal e.g.nabh 4 + = EAY!

11 11 Two-phase synthesis of MPC s toluene TOA + stir TOA + - AuCl 4 - Alkane thiol Au + - thiol NaBH 4 Au thiol nanoparticle AuCl 4 - NaBH 4 Aqueous phase Thiols(-H ) have a strong tendency to form strong bonds with Au. TOA + =tetraoctyl ammonium ion, the phase transfer catalyst Product: 2 nm gold nanoparticles with e.g. hexanethiol protective layer MPC=monolayer protected particle Au TEM-picture -size Brust et. al. J. Chem. oc. Chem. Comm. (1994) 801

12 12 Functionalised nanoparticles Thiols can have different functional groups (X) Au X X Au X X A thiol can be replaced by another with a ligand exchange reaction. In short, alkanethiol stabilised particles are mixed in solution with thiols with desired functional groups and stirred for e.g. 24 hours. Porphyrin-functionalised gold particle. Cormode et al. J. Inorg. Organomet. Polym. 18 (2008) 32

13 13 Different shapes of nanoparticles Gold nanorods TEM image of Gold nanorods 50 nm Perez-Juste et al. Coord. Chem. Rev. 249 (2005) 1870

14 Tuning the properties of nanoparticles 14 The properties of metal nanoparticles can be affected by ize and monodispersity of the particles Metal Metals can be coated by other metals to form core-shell nanoparticles Bimetallic particles tabiliser Hydrophobicity/hydrophilicity Functional groups Versatile building blocks for different applications!

15 A Optical properties of spherical 15 gold nanoparticles C 12 -Au max = 529 nm in toluene max = 580 nm in film in toluene 2 L films (100%) (nm) Above: Aggregation changes the absorbance i.e. the colour of the nanoparticles from red to blueish. This is used in many sensor applications. Left: imilar particles with different sizes have Different colours i.e. The size effects the plasmon resonance. There is absorption due to the collective motion of the conduction electrons. This is called plasmon resonance of the nanoparticles. Part of the exctinction seen in the spectrum on the left is due to this absorption, part is due to scattering of the small particles.

16 Changing the optical properties of rod-like nanoparticles 16 Absorption also at longer wavelenghts. The absorption can be tuned by the rod aspect ratio Lenght/diameter=L/d (a,b,c,d,e). Transverse and longitudinal plasmon resonance

17 Interaction of gold nanoparticles with chromophores Fluorescence quenching Energy transfer Charge transfer Fluorescence 17 Enhancement Au * Au - * + Au * 1. Chromophore is photoexcited 2. Energy is transferred from the chromophore to the gold nanoparticle 3. The excited gold nanoparticle relaxes 1. Chromophore is photoexcited 2. Charge is transferred between the chromophore and gold nanoparticle 3. Charges recombine 1. Chromophore is photoexcited 2. The electromagnetic field around gold nanoparticles affects the relaxation of the chromophore 3. Fluorescence of the chromophore is enhanced

18 A F (norm.) Example:tudy of the interaction between gold nanoparticles and porphyrins 18 Film samples on quartz plates N NH NH N Au Kotiaho, Lahtinen et al. J. Phys. Chem. C, 112 (2008) TBP=porphyrin Emission spectra Absorption spectra TBP (30%, L)/ C8Au (100%) TBP (30%, L) (nm) TBP (30%) TBP (30%)/ C8Au (100%) (nm) exc = 420 nm Gold quenches porphyrin emission

19 counts Fluorescence lifetimes bytcpc (time correlated single photon counting) TBP 2-exp fit TBP TBP/ C8-Au 3-exp fit TBP/C8-Au lamp 404 nm TBP fluorescence lifetime decreases when in contact with gold nanoparticles Quenching by energy transfer t (ns) ample t 1 (ns) P 1 (%) t 2 (ns) P 2 (%) t 3 (ns) P 3 (%) t avg (ns) 2 TBP (30%, L) TBP (30%, L)/ C8-Au (100%) 5.1 ± ± ± ± ±

20 counts (a.u.) F/F0 (at 654 nm) Dependence of quenching on distance 20 1.E+00 1.E-01 1.E-02 1.E-03 1.E-04 TCPC t (ns) Interaction mechamisns possible to study with spectroscopic methods. TBP TBP/ 5ODA/ C8Au TBP/ 3ODA/ C8Au TBP/ 1ODA/ C8Au TBP/ C8Au d (nm) F F m ODA For a film the dependence of intensity F is(n = 2 ): 1 d d from which a critical distance d o is obtained as: n d 0 = nm

21 21 Why gold for bioapplications? Gold is biocompatible. Biocompatibility of gold nanoparticles has been confirmed by various in vitro and in vivo experiments and use throughout the history of civilization Gold nanoparticles are easy to synthesize and characterize due to the presence of surface plasmon resonance band Gold is a soft acid, so it is known to bind strongly with soft bases like thiols and moderately strongly with amine functionalities. Therefore it can be functionalised in a very versatile manner. Proteins, by virtue of having cysteine and lysine residues, represent a unique ligand for binding to metal nanoparticles such as gold and alter their biological activities.

22 Bio- and medical applications of gold nanoparticles 22 Imaging Biosensors anion sensors colorimetric sensors Gold nanoparticles (GNP) as tranfers agents Photodynamic cancer therapy Plasmonic oncology

23 Gold nanoparticles as an X-Ray Contrast Agents 23 Gold nanoparticles are injected in vivo and they will enable X-ray imaging of soft tissues like kidneys. Kidneys in live mouse 60 minutes after intravenous injection of (a) gold nanoparticles or (b) iodine contrast medium ource: X-ray based computed tomography (CT) makes it possible to images even smaller tumors down to the molecular level. Dark field microscope images of neck cancer cells matching UM-A9 antibody-coated gold nanorods. cale bar: 10 μm. Ref: Popovtzer et. al. Nanolett. 8(12) (2008) 4593

24 Nanoparticles for detecting cancer cells 24 Many cancer cells have a protein, known as Epidermal Growth Factor Receptor (EFGR), all over their surface, while healthy cells typically do not express the protein as strongly. By binding the gold nanoparticles to an antibody for EFGR, anti-efgr, the nanoparticles attach themselves to the cancer cells. Then they can be detected by microscope. ource: Georgia Institute of Technology February 28th 2008

25 Gold nanoparticles in sensors -pregnancy test 25 Urine passes from the flow stick to a central reservoir containing gold nanoparticles (red) and latex microparticles (grey). Nano- and micro- particles are modified with antibodies (blue) that bind to pregnancy hormone. If pregnancy hormone (yellow) is present in urine, particles will aggregate and are unable to pass through the downstream filter. This produces a red signal in the viewing window.

26 Nanoparticles as delivery agents 26 Gold nanoparticles are capable of delivering large biomolecules, without restricting themselves as carriers of only small molecular drugs. They offer efficient recognition and delivery of biomolecules. They have shown the success in delivery of peptides, proteins, or nucleic acids like DNA or RNA. The release of the transported molecule can be triggered by internal or external stimuli. Popovtzer et. al. Nanolett. 8(12) (2008) 4593

27 Nanoparticles in cancer therapy: Photodynamic cancer therapy 27 inglet oxygen ( 1 O 2 ), a cytotoxic species, is used in photodynamic therapy of cancer. The quantum yield of free pthalocyanines PC was increased by 50% (from 0.45 to 0.65)upon conjugation to MPCs. Moreover, the conjugation promoted solubility of hydrophobic PCs in polar solvent (from toluene to ethanol). Gosh et al. Adv. Drug Deliv. Rev. 60 (2008)

28 Nanoparticles in cancer therapy: Plasmonic oncology 28 Preparation of nanoparticles that can recognise damaged cells. This is achieved by coating the nanoparticles with molecules that detect and enter cancer cells. Nanoparticles are excellent sources of heat. The shape of the nanoparticles optimises the generation of heat in response to an external light source. In principle, gold is biocompatible and is readily evacuated by body fluids. Gold nanoparticles conduct heat that can burn away cancerous cells.

29 Gold nanoparticles for treating parasites -in the future Nanoparticles (20 nm) were coated with the same antibody that body produces for the parasite Toxoplasmosis gondii Nanoparticles will attach to the parasite in the brain Nanoparticles are heated with a laser (previous slide) and the parasite dies pecific treatment Pissuwan et al. Trends in Biotechnology, 28(4)

30 30 Imaging Biosensors anion sensors colorimetric sensors FRET based sensoring Gold nanoparticles (GNP) as tranfers agents Photodynamic cancer therapy Plasmonic oncology

31 31 ummarising... You CAN buy gold nanoparticles for therapeutic purposes (probably not to be used on your own though) The easy preparation and tunable properties of biocompatible gold nanoparticles have resulted in a wide variety of applications also in the field bioscience and medicine. This research field is currently developing rapidly and new applications are likely to appear.

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