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Light Scattering When light encounters matter, matter not only re-emits light in the forward direction (leading to absorption and refractive index), but it also reemits light in all other directions. Biomedical Optics Tissue optical properties E. Göran Salerud Department Biomedical Engineering Molecule Light source This is called scattering. Light scattering is everywhere. All molecules scatter light. Surfaces scatter light. Scattering causes milk and clouds to be white and water to be blue. It is the basis of nearly all optical phenomena. Scattering can be coherent or incoherent. Spherical waves A spherical wave is also a solution to Maxwell's equations and is a good model for the light scattered by a molecule. • where k is a scalar, and • r is the radial magnitude. E( r ,t) ∝ ( E0 / r ) exp[i(kr − ω t)] A spherical wave has spherical wave-fronts. Unlike a plane wave, whose amplitude remains constant as it propagates, a spherical wave weakens. Its irradiance goes as 1/r2. Scattered spherical waves often combine to form plane waves A plane wave impinging on a surface will produce a reflected plane wave because all the spherical wavelets interfere constructively along a flat surface. The interference is measured in one direction at time and far away This way we can approximate spherical waves by plane waves in that direction, vastly simplifying the math. The mathematics of scattering The math of light scattering is analogous to that of light sources. If the phases aren t random, we add the fields: Coherent Etotal = E1 + E2 + … + En Far away, spherical wavefronts are almost flat… Usually, coherent constructive interference will occur in one direction, and destructive interference will occur in all others. Itotal = I1 + I 2 + ... + I N + ce Re{E1E2* + E1E3* + ... + EN -1EN* } I1, I2, … In are the irradiances of the various beamlets. They re all positive real numbers and add. Ei Ej* are cross terms, which have the phase factors: exp[i(qi-qj)]. When the q s are not random, they don t cancel out! If the phases are random, we add the irradiances: To understand scattering in a given situation, we compute phase delays Because the phase is constant along a wave-front, we compute the phase delay from one wave-front to another potential wave-front. If the phase delay for all scattered waves is the same, then the scattering is constructive and coherent. If it varies uniformly from 0 to 2p, then it s destructive and coherent. Incoherent Coherent constructive scattering A beam can only remain a plane wave if there s a direction for which coherent constructive interference occurs. Wave-fronts Consider the different phase delays for different paths. L1 qi qr L2 L3 L4 Scatterer Potential wave-front fi = k Li Incident wave-front Potential outgoing wave-front If it s random (perhaps due to random motion), then it s incoherent. Coherent constructive interference occurs for a reflected beam if the angle of incidence = the angle of reflection: qi = qr. Coherent destructive scattering Coherent scattering occurs in one direction, with coherent destructive scattering occurring in all others •Imagine that the reflection angle is too big. •The symmetry is now gone, and the phases are now all different. f = ka sin(qtoo big) qi qtoo big a A smooth surface scatters light coherently and constructively only in the direction whose angle of reflection equals the angle of incidence. f = ka sin(qi) Potential wave front Coherent destructive interference occurs for a reflected beam direction if the angle of incidence ≠ the angle of reflection: qi ≠ qr. Incoherent scattering: reflection from a rough surface No matter which direction we look at it, each scattered wave from a rough surface has a different phase. So scattering is incoherent, and we ll see weak light in all directions. Looking from any other direction, you ll see no light at all due to coherent destructive interference. Why can t we see a light beam? Unless the light beam is propagating right into your eye or is scattered into it, you won t see it. This is true for laser light and flashlights. This is due to the facts that air is very sparse (N is relatively small), air is also not a strong scatterer, and the scattering is incoherent. This eye sees almost no light. This eye is blinded (don t try this at home…) This is why rough surfaces look different from smooth surfaces and mirrors. To photograph light beams in laser labs, you need to blow some smoke into the beam… On-axis vs. off-axis light scattering Off-axis light scattering: scattered wavelets have random relative phases in the direction of interest due to the often random place-ment of molecular scatterers. Forward (on-axis) light scattering: scattered wavelets have nonrandom (equal!) relative phases in the forward direction. Randomly spaced scatterers in a plane Incident wave Off-axis scattering is incoherent when the scatterers are randomly arranged in space. Path lengths are random. Light scattering regimes Particle size/wavelength ~1 Large Rayleigh-Gans Scattering Mie Scattering Totally reflecting objects Geometrical optics Air Rayleigh Scattering Relative refractive index Large ~1 ~0 For large particles, we must first consider the fine-scale scattering from the surface microstructure and then integrate over the larger scale structure. If the surface isn t smooth, the scattering is incoherent. If the surfaces are smooth, then we use Snell s Law and angle-of-incidenceequals-angle-of-reflection. Incident wave Forward scattering is coherent— even if the scatterers are randomly arranged in space. Path lengths are equal. Scattering from particles is much stronger than that from molecules There are many regimes of particle scattering, depending on the particle size, the light wave-length, and the refractive index. Rainbow This plot considers only single scattering by spheres. Multiple scattering and scattering by non-spherical objects can get really complex! Then we add up all the waves resulting from all the input waves, taking into account their coherence, too. Scattering • The light scattered by a tissue has interacted with the ultrastructure of the tissue. • Tissue ultrastructure extends from membranes to membrane aggregates to collagen fibers to nuclei to cells. • Photons are most strongly scattered by those structures whose size matches the photon wavelength. Scattering possibilities Scattering • Scattering provides feedback during therapy. 10 µm cells nuclei • during laser coagulation of tissues, the onset of scattering is an observable endpoint mitochondria • Scattering has diagnostic value. 1 µm lysosomes, vesicles • the density of lipid membranes in the cells, the size of nuclei, the presence of collagen fibers, the status of hydration in the tissue. 0.1 µm striations in collagen fibre macromolecular aggregates 0.01 µm Scattering coefficient membranes Scattering Coefficient Scattering coefficient Scattering cross-section • The scattering coefficient (µs) describes a medium containing many scattering particles and is defined as: • The scattering cross-section (ss) is defined as: µs = rss s Where, ss is the scattering cross-section (cm2) rs is a volume density (cm-3) Where, s s = Qs A s Qs is the scattering efficiency (can be calculated from Mie theory) As is the area of the scatterer (cm2) Scattering Coefficient Scattering coefficient µs Reduced scattering coefficient efficiency abs. crosssectional area (µs’) • Lumped property incorporating the scattering (µs) coefficient and the anisotropy factor (g): σ s = Qs A geometrical area [cm 2 ] = [−][cm 2 ] µs ' = µs (1 - g ) Where, µs is the scattering coefficient (cm-1) σ s = Qs A A geometrical cross-section effective cross-section ECE532 Biomedical Optics ©Steven L. Jacques, Scott A. Prahl Oregon Graduat Institute Mean Free Path Scattering coefficient µs µs = ρ sσ s ls = scattering crosssectional area density The probability of transmission T of the photon without redirection by scattering after a pathlength L is: • µs • ls • Most tissues 1 µs 100 cm-1 = 0.1 mm µ s > 50 cm-1 (prostate) < 1000 cm-1 (tooth emanel) T = exp[−µsL] 24 Anisotropy g Scattering Phase Function azimuthal angle y deflection angle q Differential scattering cross section: scattering in direction s from input direction s’ dσ s (ŝ, ŝ ') dΩ y The angular dependence of scattering is q p(ŝ ⋅ ŝ ') = 4π dσ s (ŝ ⋅ ŝ ') σ s +σ a dΩ cos(q) photon trajectory scattering event • Often the scattering phase function does not depend on input direction: p(θ) • p(θ) describes the probability of a photon scattering into a unit solid angle, relative to the original photon trajectory • p(θ) has historically been called the scattering phase function 26 Scattering Anisotropy The proper definition of anisotropy (g) is the expectation value for cos (θ): Effectiveness of Scattering g≡ ∫ p(ŝ ⋅ ŝ ')ŝ ⋅ ŝ 'dΩ ∫ p(ŝ ⋅ ŝ ')dΩ ∫ • The proper definition of anisotropy (g) is the expectation value for cos (q): 1 π p(θ )cosθ 2π sin θ dθ 0 = ∫ p(cosθ )cosθ d(cosθ ) -1 1 π where ∫ 0 p(θ )2π sin θ dθ = 1 Expression for anisotropy Sometimes also written in terms of cos(θ) g =< cos (θ ) = Scattering Coefficient where ∫ p(cosθ )d(cosθ ) = 1 −1 g =< cosq > Scattering Anisotropy • Anisotropy is a measure of forward direction retained after a single scattering event (mean value of cos(θ)) " −1 Total backward scattering $ g=# 0 $ 1 Total forward scattering % Scattering function • The angular dependence of scattering is called the scattering function, p(q) which has units of [sr-1] and describes the probability of a photon scattering into a unit solid angle oriented at an angle relative to the photons original trajectory. BiologicalTissues,0.65<g>0.95 29 Scattering function Scattering functions • Plotting p(q) indicates how photons will scatter as a function of q in a single plane of observation • Plotting p(q) 2psinq indicates how photons will scatter as a function of the deflection angle q regardless of the azimuthal angle y, in other words integrating over all possible y in an azimuthal ring of width dq and perimeter 2psinq at some given q . ECE532 Biomedical Optics ©Steven L. Jacques, Scott A. Prahl Oregon Graduate Institute Isotropic scattering Albedo π 1 p(θ ) = , such that 4π p(ŝ ⋅ ŝ ') = ∫ p(θ )2π sinθdθ = 1 4π dσ s (ŝ ⋅ ŝ ') σ s +σ a dΩ Fraction of light energy incident on a scatterer or absorber from direction s that gets scattered into direction s prime. 0 g=0 ∫ p(ŝ ⋅ ŝ ')dΩ = σ 4π µs = + σ µ s a s + µa Albedo, in tissue can range from 0.3 to 0.99 depends on wavelength Ratio of scattering versus total attenuation Scattering in tissue is dominant! 34 Henyey-Greenstein scattering function Henyey-Greenstein Heyney Greenstein scattering phase function is an analytical expression which mimics the angular dependence of light scattering by small particles and is based on the anistropy factor g (used for Monte Carlo Simulations) • The function allows the anisotropy factor, g to specify p(θ) such that <cos(θ)> returns the same value of g • This function is useful in approximating the angular dependence of single scattering events in biological tissue π 1 1− g 2 p(θ ) = , such that 2 4π (1+ g − 2g cosθ )3/2 ∫ p(θ )2π sinθ dθ = 1 0 π and ∫ p(cosθ )d(cosθ ) = 1 • The function does not represent true scattering phase functions very well but it is a good average approximation 0 p(θ ) = 1 1− g 2 , such that 2 2 (1+ g − 2g cosθ )3/2 π ∫ p(cosθ )d(cosθ ) = 1 0 π and ∫ p(θ )2π sinθ dθ = 1 0 36 Different anisotropy values Reduced scattering coefficient • µs' = µs(1 - g) [cm-1] • The purpose of µs' is to describe the diffusion of photons in a random walk of step size of 1/µs' [cm] where each step involves isotropic scattering. • This occurs if there are many scattering events before an absorption event, i.e., µa << µs'. • This situation of scattering-dominated light transport is called the diffusion regime and µs’ is useful in the diffusion regime which is commonly encountered when treating how visible and near-infrared light propagates through biological tissues. ECE532 Biomedical Optics ©Steven L. Jacques, Scott A. Prahl Oregon Graduate Institute Example, reduced scattering coefficient Importance of light scattering • Important because g = <cos(q) = 0.90 • light propagation is affected by the tissue optical properties, 26o • the physiological condition or state of single cells or tissues is expressed through (but not exclusively) changes in cell size or refractive index, <q> ≈ one mfp´ • changes in refractive index or cell size influence the optical properties. µs = µs(1-g) = 0.1 mfp = 1/µs mfp = 1/µs ten mfp Therefore measurements or analysis of scattering provide information about the tissue. Derived Characteristic properties • Four important properties • Cross section • • absorption • scattering • extinction Cross sections Scattering cross section σs = 4π σt 4π ∫ Albedo p(s,i)dω 4π W0 = Absorption cross section σa Angular dependence • ∫ σ d dω = scattering phase function σs σ = s σs + σa σt Note: W0 is close to zero for most tissues Back-scatter cross section σ b = 4 πσ d (−i,i) Extinction cross section σt = σs + σa σ b = differential scattering cross section Scattering cases • Size parameter x = 2πa /( l/nmed) • Refractive index ratio nr = np/nmed • Rayleigh approximation • the particle a dipole, strength related to its volume • valid if a<<1 or until a is 5% of l • proportional to l-4 Rayleigh scattering • Scattered intensity I s ( R,θ ) = (1+ cos 2 θ ) • k4 2 α Ii 2 2R where k=angular wavenumber and nr=refractive index ratio k= 2π nmed λ nr = np nmed Rayleigh scattering • • Scattering cross-section σs = Rayleigh scattering 8π k 4 2 α 3 Polarizability of a sphere n2 − 1 3 α = 2r a nr + 2 Scattering cross-section 8π a 2 x 4 nr2 − 1 σs = , x = ka 3 nr2 + 2 2 • • • Size parameter x = ka Scattering efficiency 8x 4 nr2 − 1 Qs = 3 nr2 + 2 2 • Diameter of sphere: 2a = 20 nm • Wavelength in vacuum: l = 400 nm • Refractive index of sphere: ns = 1.57 • Refractive index of background: nb = 1.33 • Specific weight of sphere: rs = 1.05 g/cm3 • Specific weight of background: rb = 1.00 g/cm3 • Concentration of spheres in background by weight: Cwt = 10-5 • ss = 2.15x10-20 m2 • Qs = 6.83x10-5 Mie theory model for tissue optical properties • Mie theory describes the scattering of light by particles, with refractive index (np) that differs from the refractive index of its surroundings (nmed). • The dipole re-radiation pattern from oscillating electrons in the molecules of such particles superimpose to yield a strong net source of scattered radiation. • • Also, the re-radiation patterns from all the dipoles do not cancel in all but the forward direction of the incident light as is true for homogeneous medium, but rather interfere both constructively and destructively in a radiation pattern. Hence, particles "scatter" light in various directions with varying efficiency. Mie • Mie's classical solution is described in terms of two parameters, nr and x: • the magnitude of refractive index mismatch between particle and medium expressed as the ratio of the n for particle and medium, nr = np/nmed • a the size of the surface of refractive index mismatch which is the "antenna" for re-radiation of electro-magnetic energy, expressed as a size parameter (x) which is the ratio of the meridional circumference of the sphere (2pa, where radius = a) to the wavelength (l/nmed) of light in the medium, x = 2pa /( l/nmed) Mie calculations MIE scattering • A Mie theory calculation will yield the efficiency of scattering which relates the cross-sectional area of scattering, ss to the true geometrical cross-sectional area of the particle, A = pa2 {Q s,p(θ )} = Mie( np 2πa , ) n med λ /n med σ s = Q s πa 2 • ss = QsA • Finally, the scattering coefficient is related to the product of scatterer number density rs, and the cross-sectional area of scattering, ss µs = rsss π g= ∫ p(θ )cosθ ⋅ 2π sinθdθ 0 π ∫ p(θ ) ⋅ 2π sinθdθ 0 The math of Mie scattering Scattering matrix • Describes the relationship between incident and scattered electric field components perpendicular and parallel to the scattering plane as observed in the "far-field” ⎡E lls ⎤ exp(−ik(r − z)) ⎡S2 S3 ⎤⎡E lli ⎤ ⎢ ⎥= ⎢ ⎥⎢ ⎥ ikr ⎣E ⊥s⎦ ⎣S4 S1 ⎦⎣E ⊥i ⎦ • S is known as the "amplitude scattering matrix” • The total field (Etot) depends on the incident field (Ei), the scattered field (Es) , and the interaction of these fields (Eint). If one observes the scattering from a position which avoids Ei, then both Ei and Eint are zero and only Es is observed. Scattering matrix • For the "far-field” observation of Es at a distance r and particle of diameter d such that: kr >> nc2 2π k= λ d nc = λ • Then the scattering elements S3 and S4 are equal to zero. Measured scattering matrix • Practical experiments measure intensity, I =< EE * >= (1/2)a 2 ,where E = aexp(−iδ ) ⎡ Ills ⎢ ⎢⎣ I⊥ s ⎡ S ⎤ 2 ⎥ = constant ⎢ ⎢ 0 ⎥⎦ ⎢⎣ 2 0 S1 2 Stokes vector Angular scattering pattern of polarized light ⎤⎡ ⎥ ⎢ Illi ⎥ ⎢ I⊥ i ⎥⎦ ⎣ Angular patterns of Mie scattering Click here for Mie calculator http://omlc.ogi.edu/calc/mie_calc.html ⎤ ⎥ ⎥⎦ Scattered light intensity for random polarised Mie theory calculation of anisotropy (g) π 2 2 1 1 S11 (θ ) = S1 (θ ) + S2 (θ ) 2 2 where S11 (θ ) = first element in Mueller matrix g= ∫ S (θ ) cos(θ )2π sin(θ )dθ 11 0 π ∫ S (θ )2π sin(θ ) dθ 11 I s = S11 * I i 0 p(θ ) = Mueller Matrix, a 4x4 matrix which relates an input vector of Stokes parameters (Ii, Qi, Ui, Vi) describing a complex light source and the output vector (Is, Qs, Us, Vs) describing the nature of the transmitted light. S11 (θ ) π ∫ S (θ )2π sin(θ ) dθ 11 0 Scattering vs wavelength Mie scattering from cellular structures • Soft tissue optics are dominated by the lipid content of the tissues. • Mie theory provides a simple first approximation to the scattering of soft tissues • The approximation involves a few assumptions: • Assume the refractive index of the lipid membranes of cells is 1.46, based on the reported • Assume the refractive index of the cytosol of cells is 1.35, based on the reported value for cellular cytoplasms • Assume the lipid content of soft tissue is about 1-10% (fv = 0.02-0.10). • Let's choose fv = 0.02 for this example to match the value for several typical soft tissues such as lungs, spleen, prostate, ovary, intestine, liver, arteries, to name a few. • Assume all the lipid is packaged as small spheres of various sizes whose number density maintains a constant volume fraction fv. • Ignore the interference of scattered fields from particles which can alter the apparent scattering properties based on isolated particles. Scattering vs wavelength Scattering Assumptions • The optical properties of the particle are completely described by the refractive index • The medium in which the particles are embedded is considered to be homogeneous • Only the interactions of a single particle with light of arbitrary wavelength are considered • The total scattered field is merely the sum of the fields scattered by each particle (i.e. the particles do not affect each other). • For a collection of particles, the number of particles is large and their separations are random ECE532 Biomedical Optics ©Steven L. Jacques, Scott A. Prahl Oregon Graduate Institute 62 Scattering vs wavelength Scattering vs wavelength ECE532 Biomedical Optics ©Steven L. Jacques, Scott A. Prahl Oregon Graduate Institute ECE532 Biomedical Optics ©Steven L. Jacques, Scott A. Prahl Oregon Graduate Institute Summary optical properties Summary • Optically tissue may be characterized by its • • • scattering, refractive index, and absorption. • The scattering arises from • cell membranes, cell nuclei, capillary walls, hair follicles... • The absorption arises from • visible and NIR wavelengths (400 nm - 800 nm); • » hemoglobin and melanin, • IR wavelengths; • » water and molecular vibrational/rotational states. A survey of all tissue types has not been completed. A survey would only be a guideline since subject to subject variation and normal vs diseased variation apparently are sufficiently significant to warrant ad hoc measurements on any particular tissue site of interest.Other tissues may present different types of scattering than just the simple "soft tissue" and "dermis" models. • Muscles have myoglobin and actin-myosin fibers. • Brain has myelin sheaths around nerves. • Fatty tissues are distinct in their properties. The scattering by the nucleus has been ignored here, but is a potential scattering site of clinical importance. Wavelength dependence in tissue Wavelength dependence in tissue • Absorption and scattering decreases as a function of wavelength • Ratio of scattering to absorption coefficient increases with the wavelength www.liu.se