Nanoparticles for in-vitro and in-vivo biosensing and imaging

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6 CONTENTS 6.15.2 Olympus BX51 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 242 6.16 The non-descanned detection system . . . . . . . . . . . . . . . . . . . . . 243 6.16.1 The ND-unit design . . . . . . . . . . . . . . . . . . . . . . . . . . 244 6.17 Detectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 245 6.17.1 Single Photon Avalanche Diode (SPAD) . . . . . . . . . . . . . . . 246 6.18 Dynamic Light Scattering:optical system . . . . . . . . . . . . . . . . . . . 248 6.18.1 Softwares . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 250 6.19 TimeHarp and Symphotime softwares . . . . . . . . . . . . . . . . . . . . 252
Introduction In the last two decades several groups have investigated the changes of chemical and physical properties of materials with size in nanometric scales. These studies have highlighted a number of possible applications for nanostructures, which are now employed, for example, in biology and medicine for imaging, disease detection, diagnosis, sensing and therapy. Noble metal (especially gold and silver nanoparticles) are particularly versatile for these applications due to the phenomenon known as surface plasmon resonance (SPR), an in-phase oscillation of all the conduction band electrons that resonates with the light wave electric field. The resonance frequency depends on the size, shape, orientation and dielectric constant of the nanoparticle. This coupling of SPR with the electromagnetic field leads to a great enhancement of all the nanoparticle radiative properties, such as absorption and scattering. The extinction cross section of these nanoparticles is 10 5 -10 6 larger than that of organic dye and in contrast to common molecular chromophores they are extremely photostable and, depending on the shape, they convert efficiently light into heat. The SPR, tunable in the visible (for spherical NPs) and near-infrared region (for anisotropic Nps) of the electromagnetic spectrum, can also interact, for gold nanoparticles a few nanometers in size, with the fluorescence emission of dyes and substantially modify their brightness and excited-state lifetime. Depending on the fluorophores-NP distance and the NPs anisotropy one can obtain fluorescence enhancement or quenching. In both cases we expect that any change in the dielectric constant of the NP surface, induced, for example, by a biorecognition process that occurs on the surface itself, can produce a change in the emission properties of the fluorophores. SPR effect becomes also particularly important when combined with two-photon excitation (TPE), which consists in the simultaneous absorption of two photons, each carrying about half the energy necessary to excite the molecule. This excitation technique has been particularly useful in the context of this work because it affects greatly the luminescence quantum yield of anisotropic NPs: due to non-linear phenomena such as the TPE, the luminescence (TPL) intensity is enhanced (when coupled with an appropriate 7
Page 1 and 2: UNIVERSITÀ DEGLI STUDI DI MILANO B
Page 3 and 4: CONTENTS 3 3.2 Dynamic Light scatte
Page 5: CONTENTS 5 5.15.2 Spectral characte
Page 9 and 10: Chapter 9 then we have applied the
Page 11 and 12: Chapter 1 Metal nanoparticles 1.1 N
Page 13 and 14: Chapter 1 13 carry out reactions in
Page 15 and 16: Chapter 1 15 donor and the metal at
Page 17 and 18: Chapter 1 17 Figure 1.4: Surface pl
Page 19 and 20: Chapter 1 19 where the scattering a
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Page 23 and 24: Chapter 1 23 easily be seen from eq
Page 25 and 26: Chapter 1 25 The first is equal to
Page 27 and 28: Chapter 1 27 dipole approximation a
Page 29 and 30: Chapter 1 29 used for disease or ca
Page 31 and 32: Bibliography [1] Nanotoxicology: na
Page 33 and 34: Chapter 1 33 [30] Bohren CF; Huffma
Page 35 and 36: Chapter 2 Principles 2.1 Basics of
Page 37 and 38: Chapter 2 37 A direct consequence o
Page 39 and 40: Chapter 2 39 be considerably more c
Page 41 and 42: Chapter 2 41 Figure 2.5: Effect of
Page 43 and 44: Chapter 2 43 2.4 Two-photon excited
Page 45 and 46: Chapter 2 45 m = λV MI hc 2 N A (2
Page 47 and 48: Chapter 2 47 P SF OP E (u, v) = |h(
Page 49 and 50: Chapter 2 49 renditions of the spec
Page 51 and 52: Bibliography [1] Lakowicz J.R. In p
Page 53 and 54: Chapter 2 53 [29] Born M. and Wolf
Page 55 and 56: Chapter 3 55 3.2 Dynamic Light scat
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Chapter 3 57 where n is the refract
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Chapter 3 59 with g V H (t) ∝ β
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Chapter 3 61 κ m (Γ) = dm K(−τ
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Chapter 3 63 that plagues conventio
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Chapter 3 65 set equal to the new p
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Chapter 3 67 the molecules. In orde
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Chapter 3 69 number of molecules in
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Chapter 3 71 γ 1 G(τ) = V ex 〈C
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Chapter 3 73 Figure 3.2: Auto-corre
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Chapter 3 75 electronic distortions
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Chapter 3 77 Figure 3.5: left panel
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Chapter 3 79 p 1 (k, V 0 , ɛ) = 1
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Chapter 3 81 3.9.3 Photon Moment An
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Chapter 3 83 Figure 3.6: Schematic
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Bibliography [1] Chu B. In Laser Li
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Chapter 3 87 [27] Eigen S.M. and Ri
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Chapter 3 89 [53] Chen Y., Tekmen M
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Chapter 4 91 • L.Sironi, S.Freddi
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Chapter 4 93 die. Mutations in the
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Chapter 4 95 arm of chromosome 17.
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Chapter 4 97 shown that one of the
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Chapter 4 99 DNA replication while
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Chapter 4 101 4.2). The ratio R = [
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Chapter 4 103 The polydispersity of
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Chapter 4 105 The fitting of the FC
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Chapter 4 107 the streptavidin (4.5
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Chapter 4 109 29 nm for the 10 nm s
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Chapter 4 111 A 1 Γ 1 (nm) λ 1 (n
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Chapter 4 113 Figure 4.8: (Left: Fl
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Chapter 4 115 been systematically i
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Chapter 4 117 Figure 4.12: Left: Me
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Chapter 4 119 at the basis of the o
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Chapter 4 121 presence of p53, comp
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Chapter 4 123 Figure 4.17: The figu
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Chapter 4 125 from probably aspecif
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Chapter 4 127 Figure 4.21: Panel A:
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Bibliography [1] Anker J. N.; Hall
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Chapter 4 131 [47] Das P.; Metiu H.
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Chapter 5 133 In this chapter the c
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Chapter 5 135 Figure 5.1: Left:Symm
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Chapter 5 137 5.3 Experimental deta
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Chapter 5 139 back aperture of the
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Chapter 5 141 5.4.2 Transmission El
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Chapter 5 143 For anisotropic branc
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Chapter 5 145 coefficient D is then
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Chapter 5 147 Figure 5.9: ACF of th
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Chapter 5 149 measured through a ve
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Chapter 5 151 objective in epifluor
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Chapter 5 153 The fitting of FCS cu
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Chapter 5 155 Sample Population (%)
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Chapter 5 157 5.8 Concentration eva
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Chapter 5 159 Figure 5.17: Emission
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Chapter 5 161 5.12 Dependence of TP
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Chapter 5 163 Figure 5.22: Histogra
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Chapter 5 165 5.13 Cellular uptake
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Chapter 5 167 Figure 5.29: NPs obta
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Chapter 5 169 Figure 5.33: Autofluo
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Chapter 5 171 5.14 Photothermal eff
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Chapter 5 173 Figure 5.37: ζ-poten
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Chapter 5 175 Temperature [ ◦ C]
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Chapter 5 177 In figure 5.39 the fl
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Chapter 5 179 P [mW] < τ > [ns] 10
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Chapter 5 181 The dependence of the
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Chapter 6 183 Currently, only alumi
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Chapter 6 185 tact organ studies, a
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Chapter 6 187 with liquid flow chan
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Chapter 6 189 Figure 6.1: DC in clo
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Chapter 6 191 • New contrast agen
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Chapter 6 193 experiments by flowin
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Chapter 6 195 it has discontinuitie
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Chapter 6 197 P t. rection. If all
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Chapter 6 199 Figure 6.6: Commonly
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Chapter 6 201 Figure 6.9: 3D and 2D
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Chapter 6 203 Figure 6.11: (A) NK c
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Chapter 6 205 are somehow confined,
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Chapter 6 207 Figure 6.15: Distance
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Chapter 6 209 Figure 6.18: (A)The s
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Chapter 6 211 Figure 6.21: The figu
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Chapter 6 213 Figure 6.24: Paramete
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Chapter 6 215 of the natural killer
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Chapter 6 217 and CD8 + T cells, is
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Chapter 6 219 6.10.1 TLRs TLRs are
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Chapter 6 221 arise from myeloid pr
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Chapter 6 223 of infection, primari
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Chapter 6 225 Figure 6.30: Lymph-no
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Bibliography [1] A. Diaspro, G. Chi
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Chapter 6 229 [25] E. O. Long. Regu
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Chapter 6 231 [47] S. H. Wei, M. J.
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Chapter 6 233 [72] S. Fossum and W.
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Chapter 235 mode-locked cavity. Thi
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Chapter 237 Figure 6.34: Energy lev
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Chapter 239 Figure 6.37: Prisms seq
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Chapter 241 part of the TE300 and t
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Chapter 243 Figure 6.42: The Olympu
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Chapter 245 a f=100 mm bispherical
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Chapter 247 are just events’ dete
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Chapter 249 smaller quartz cell (He
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Chapter 251 needed to a linear corr
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Chapter 253 Figure 6.48: Channel ar
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Chapter 255 ming for burst height a
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Chapter 257 • M. Caccia, T. Gorle
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