3. Theory of one, two and N plasmonic epitopes
We start our analysis with a single plasmonic epitope on the equator of a microspheroid (
Fig. 3
) in an attempt to verify the results of Zhu
et al. [
11J. Zhu, Ş. K. Ozdemir, Y. F. Xiao, L. Li, L. He, D. R. Chen, and L. Yang, “On-chip single nanoparticle detection and sizing by mode splitting in an ultrahigh-Q microresonator,” Nat. Photonics 4(1), 46–49 (2010).
] before moving on to the necklace. We concentrate theoretically on the equatorial mode of an oblate axisymmetric microcavity. Without the plasmonic epitope or significant roughness we have a bare cavity in which two counterpropagating equatorial WGMs have identical resonance frequencies; they are degenerate. Since the microcavity is a high
Q system, the modes of this system can be considered to be quasi-normal modes [
21E. S. C. Ching, P. T. Leung, and K. Young, “The role of quasinormal modes,” in Optical Processes In Microcavities, R. K. Chang, A. J. Campillo, eds. (World Scientific, 1996), pp. 1–75.
]. This allows for quantum analogs such as photonic atoms [
22S. Arnold, “Microspheres, photonic atoms, and the physics of nothing,” Am. Sci. 89, 414–421 (2001).
]. Such analogs compel one to borrow the theoretical machinery of quantum mechanics in order to describe high
Q optical systems. For bound nanoplasmonic epitope perturbing a WGM, degenerate perturbation theory can be used to predict the eigenvalues and eigenstates of the system.
Fig. 3 Illustration of a WGM ring resonator with one epitope.
The oblateness of the spheroid lifts the
degeneracy of a sphere allowing the equitorial mode having a principle quantum number
s, angular momentum quantum number
, and a two fold degeneracy
to be excited through guided wave excitation. This equitorial WGM (
Fig. 3) provides the simplest model for which a first order perturbation will not couple radial modes having different principle quantum numbers since their frequencies do not cross. First, we start to interpret the signal in the presence of a plasmonic epitope attached to the WGM ring (equator) at an angle
, as shown in
Fig. 3; then we apply this theory to two epitopes and N epitopes, respectively. The N epitopes will first be considered as random and then periodic.
Our theory will apply the approach of Teraoka and Arnold [
23I. Teraoka and S. Arnold, “Resonance shifts of counterpropagating whispering-gallery modes: degenerate perturbation theory and application to resonator sensors with axial symmetry,” J. Opt. Soc. Am. B 26(7), 1321–1329 (2009).
]. We will briefly review the results of this paper before applying it to the case of periodic epitopes.
First we start with the bare cavity framework. By separating the Helmholtz equation the azimuthal differential equation for the equitorial TE mode, which is almost entirely polarized along the polar direction (
) has the form:
where
r is radial variable (~radius of the ring),
k is the wave vector and
is the electric field. The solutions of the above Eq. take the form
The orbital boundary condition,
where integer
m guarantees that
m =
kr. Bare WGM microcavities typically support both clockwise (
cw) and counterclockwise (
ccw) equitorial modes, which for the same
m are degenerate in frequency. Using
Eqs. (2) and
(3), orthogonal basis vectors for these
ccw and
cw equatorial traveling waves are
respectively, where
is the semi-classical field amplitude associated with a single photon. Since our interest is in the TE equatorial mode,
[
23I. Teraoka and S. Arnold, “Resonance shifts of counterpropagating whispering-gallery modes: degenerate perturbation theory and application to resonator sensors with axial symmetry,” J. Opt. Soc. Am. B 26(7), 1321–1329 (2009).
]. The dielectric function of the cavity is
and consequently the energy in either travelling wave
, where the 2 accounts for an equal amount of electric and magnetic energy in the cavity, the matrix element requires integration over the cavity volume, and the usual conjugate relationship between the bra and ket is maintained. In order to return to a familiar result we will at first suppose that a perturbation to the cavity maintains the travelling wave. Having to polarize an added Rayleigh particle on the equator at position
with excess polarizability having a real part
corresponds to a dielectric perturbation
. This perturbation costs the cavity energy
[
3S. Arnold, M. Khoshsima, I. Teraoka, S. Holler, and F. Vollmer, “Shift of whispering-gallery modes in microspheres by protein adsorption,” Opt. Lett. 28(4), 272–274 (2003).
], for which the frequency shift of either travelling wave is
After the original presentation of
Eq. (5) [
3S. Arnold, M. Khoshsima, I. Teraoka, S. Holler, and F. Vollmer, “Shift of whispering-gallery modes in microspheres by protein adsorption,” Opt. Lett. 28(4), 272–274 (2003).
], this first order perturbation in frequency of either of the traveling wave states has become known in the literature as
g [
11J. Zhu, Ş. K. Ozdemir, Y. F. Xiao, L. Li, L. He, D. R. Chen, and L. Yang, “On-chip single nanoparticle detection and sizing by mode splitting in an ultrahigh-Q microresonator,” Nat. Photonics 4(1), 46–49 (2010).
,
24A. Mazzei, S. Götzinger, L. S. Menezes, G. Zumofen, O. Benson, and V. Sandoghdar, “Controlled Coupling of Counterpropagating Whispering-Gallery Modes by a Single Rayleigh Scatterer: A Classical Problem in a Quantum Optical Light,” Phys. Rev. Lett. 99(17), 173603 (2007).
].
Equation (5) is in a conveniently simple form by keeping Planck’s constant in place. However, because of our single photon approach
is simply a place saver for the energy in the cavity divided by frequency. With
converted in this way
Eq. (5) can be evaluated using our basis states in
Eq. (4) and the position
of the plasmonic epitope with the result [
3S. Arnold, M. Khoshsima, I. Teraoka, S. Holler, and F. Vollmer, “Shift of whispering-gallery modes in microspheres by protein adsorption,” Opt. Lett. 28(4), 272–274 (2003).
]:
It is important to understand that had we constructed our theory for N
p photons
g would not change since the intensities in the numerator and denominator scale together with N
p. In what follows we will stick with the form in
Eq. (5) by including
. However, our description currently is not complete since the dielectric perturbation can couple the traveling wave states.
Coupling requires interaction between the
ccw and
cw states through degenerate perturbation theory. For a two state system one has to solve an eigenvalue equation involving a 2x2 matrix
where
accw and
acw are amplitudes for being in a superposition state
In the absence of an off-diagonal term we return to
Eq. (5) as expected. Unique solutions are guaranteed by setting the determinant of the secular equations represented by
Eq. (7) equal to zero.
The diagonal terms can be written as
, whereas the off-diagonal terms are quickly evaluated from
Eqs. (4) since
and consequently the upper rt. element is
, since the perturbation is at azimuthal angle
. The matrix element on the lower left is just the conjugate of this,
. With these inputs
Eq. (9) becomes
where
is the optical phase at the epitope. The eigenvalues obtained from the determinant are
in agreement with Zhu
et al. [
11J. Zhu, Ş. K. Ozdemir, Y. F. Xiao, L. Li, L. He, D. R. Chen, and L. Yang, “On-chip single nanoparticle detection and sizing by mode splitting in an ultrahigh-Q microresonator,” Nat. Photonics 4(1), 46–49 (2010).
]. The epitope clearly splits the degeneracy, and the magnitude of the splitting 2|
g| depends only on the value of
g. If
g is a negative number, as it would be for an epitope having a polarizability which is positive,
is larger than
. As we go forward the subscript + on the frequency shift will be assigned to the state maintaining the higher frequency for g < 0 (i.e.
).
So far we have been assuming quasi-normal modes. Losses can cause the modal line widths to overlap, in which case one expects to measure a shift of a single feature that is the average of frequency shift of the two states. In order to look at the wave function amplitudes of the mixed states (
i.e., the location of the nodes and anti-nodes in the relation to the epitopes), we need to determine the eigenvectors in
Eq. (10). The solution of the following equation provides the required eigenvectors,
For simplicity, the epitope is assumed to be on the
X-axis (i.e.,
) then this Eq. is reduced as follows,
The eigenvalues remain the same (
). The relation between eigenvectors corresponding to the eigenvalues
and
, respectively are
and
. After inserting these relations in the
Eq. (8) (using
), and normalizing the eigen vectors to a single photon (
i.e.,
), we obtain the following new states.
and
It is clear that these wave functions are mutually orthogonal. In other words, in the presence of an epitope, the wave functions redistribute themselves in such a way that one standing wave mode
has an antinode at the epitope thereby maximizing the interaction; a symmetric standing wave (SSW), resulting in a frequency shift 2
g. It should be noted that the amplitude squared of the SSW state is twice that of either travelling wave state consistent with the factor of 2 increase in frequency shift. The other state
minimizes reactive interaction: an antisymmetric standing wave (ASW), resulting in no shift.
If another epitope is attached to the ring at an angle
, then the eigenvalues in the presence of the two epitopes are derived from the determinant
where
represents the optical phase of the wave at the second epitope. The eigenvalues of the system are
The total magnitude of the splitting (
) in the presence of two epitopes is
. This is consistent with the results in Ref [
25J. Zhu, Ş. K. Özdemir, L. He, and L. Yang, “Controlled manipulation of mode splitting in an optical microcavity by two Rayleigh scatterers,” Opt. Express 18(23), 23535–23543 (2010).
] and reveals that splitting magnitude is not only determined by the size, shape, and refraction index of the epitope, but also is strongly dependent on the phase difference between them (
i.e., relative positions). If the second epitope is identical to the first epitope (
g1 =
g2 =
g), then the total magnitude of the splitting is 4|
g|
. This clearly indicates that the mode splitting disappears when the phase difference is π/2, which results from the fact that the two epitopes exert the same influence on each of the two standing wave modes. However, if the phase difference between any consecutive epitopes is π, this gives rise to the maximum splitting 4|g|. In order to get the eigenvectors, for simplicity
and
are assumed to be 0 and π. The solution of the following equation provides the required eigenvectors;
The relation between eigenvectors corresponding to the eigenvalues
and
, respectively are
and
. By substituting these relations in the
Eq. (8), we get the same mutually orthogonal wave functions given by
Eqs. (14) and
15.
Suppose now that N epitopes are attached to the surface in random positions on the ring. Then the eigenvalues of the system are derived from the determinant
This equation can be written as
For random adsorption the ensemble average
Therefore the eigenvalues of the system are
and the magnitude of splitting is 2|
g|
. This is consistent with the results of Chantada
et al. [
26L. Chantada, N. I. Nikolaev, A. L. Ivanov, P. Borri, and W. Langbein, “Optical resonances in microcylinders: response to perturbations for biosensing,” J. Opt. Soc. Am. B 25(8), 1312–1321 (2008).
].
Having shown that our theoretical approach is consistent with the results of others we now apply it to the case of periodic epitopes. If N identical epitopes are periodically attached to the ring in the presence of an optical phase difference ΔΦ between the consecutive epitopes, then the eigenvalues of the system are given by the following equation,
where
. The eigenvalues of this system are
When
the eigenvalues are
The total magnitude of the splitting in this case is 2N|
g|. This means, when the optical phase difference between consecutive epitopes is π, then frequency shift due to each epitope adds independently. In the presence of 10 epitopes, variation in the frequency shift as a function of ΔФ is shown in
Fig. 4
. It can be seen that maximum frequency shift is 20g at ΔФ = π due to the superposition of the travelling waves, and the average shift for all possible phase differences is 10g; this is exactly the shift produced by a travelling wave. The peak that appears at π is known as the first order peak and higher order peaks appear at integer multiples of π. The peak width is inversely proportional to
N; in order to observe a significant splitting experimentally, the inter-epitopes phase difference must have an error of less than half of the peak width. If the position of each epitope is Gaussian-distributed around the correct position with standard deviation σ, the mode splitting will be reduced. For example, for an operating wavelength of 800 nm in silica, if σ/
r = 0.025 μm/40 μm, the mode splitting becomes 90% of the full splitting. For a smaller σ of 0.015 μm, the fraction rises to 96%. If one epitope is missing, then the model fails. In practice, both issues can be dealt with by quality control and post-fabrication examination. We can also view the proposed periodic epitope system as a diffraction grating, with each epitope being the grating groove. It is well known that the grating sensitivity (resolution) depends upon the total number of grooves illuminated on the surface of the grating (
N) and diffraction order (
n). Although the sensitivity (magnitude of splitting) of the proposed epitope system is dependent on
N like a diffraction grating, it is independent of
n. In other words, the magnitude of the splitting is the same for all orders.
Fig. 4 Variation of the frequency shift of the WGM in the presence of 10 epitopes adsorbed on a microcavity as a function of phase difference between the consecutive epitopes.
The eigenvectors for the
N-periodic epitope system are given by the following Equation by assuming
and
.
The relation between eigenvectors corresponding to the eigenvalues
and
, respectively are
and
, provide the same mutually orthogonal wave functions given by
Eqs. (14) and
15. The eigenstates of a system with 10 epitopes on a ring are shown in
Fig. 5
. It is clear that all epitopes are at anti-nodes (symmetric mode) in one case, resulting in the maximum frequency shift 20g. In the other case, all epitopes are at nodes (asymmetric mode), which results in no shift.
Fig. 5 Two states of a WGM ring with 10 plasmonic epitopes. Here optical phase difference between the consecutive epitopes is π.
For N periodic epitopes, that are different from each other, then the eigenvalues of the system are given by the following equation,
when
the eigenvalues of the system are
In the case of mode splitting, along with the physical properties of the epitopes, the quality factor of the microcavity also plays a major role. To observe the mode splitting requires that its magnitude be considerably larger than the line width of WGM resonance. In the case of a cavity containing
N particles, the observation of mode splitting requires
where
is the spectral width due to the scattering and absorption losses of an epitope and
is the intrinsic resonance width of the bare cavity. Thus, increasing the value of
N increases the possibility of observing the splitting.
In single dielectric nanoparticle (
e.g., bio-nanoparticle) detection, if a dielectric analyte is adsorbed to any one of the epitopes then (
N-1) epitopes have the same interaction with the microcavity (
g) and one epitope which adsorbs the analyte particle has the different interaction (
). Here it is to be noted that
is the combined interaction of single dielectric analyte particle and epitope with the microcavity. The splitting now changes to
. Similarly, if two dielectric nanoparticles are adsorbed each at the center of the dipole hot spots of two different epitopes (
i.e., position T or B in
Fig. 2), then the splitting becomes
, and so on. Here, the interaction
is proportional to the real part of effective polarizability
of a plasmonic-dielectric cluster. Suppose that an extremely small dielectric particle (
i.e., a protein molecule) is attached to a spherical plasmonic epitope, then
for transverse electric (TE) mode excitation can be estimated using the following analytical expression:
where
is the real part of the excess polarizability of the plasmonic epitope,
and
are the electric permittivities of the vacuum and relative permittivity of the surrounding medium, respectively,
is the radius of the plasmonic epitope (
e.g., solid gold sphere or gold shell structure as in
Fig. 2),
excess polarizability of the adsorbed dielectric particle, and the squared quantity on the right hand side is the intensity enhancement factor at the center of the hot spot.
Figure 6
demonstrates the advantage of the periodic plasmonic epitope system with respect to sensing. We first start the discussion of this figure by considering adsorption onto the equator of a bare resonator (lower trace), and take our analyte to be the smallest aqueous borne RNA virus [MS2, 13.6 nm radius]. This discussion will be an extension of Ref [
6V. R. Dantham, S. Holler, V. Kolchenko, Z. Wan, and S. Arnold, “Taking whispering gallery-mode single virus detection and sizing to the limit,” Appl. Phys. Lett. 101(4), 043704 (2012).
] for which the radius of the silica resonator was
and the unperturbed resonance occurs near a free space wavelength of 780nm [
6V. R. Dantham, S. Holler, V. Kolchenko, Z. Wan, and S. Arnold, “Taking whispering gallery-mode single virus detection and sizing to the limit,” Appl. Phys. Lett. 101(4), 043704 (2012).
]. To be consistent with Ref [
6V. R. Dantham, S. Holler, V. Kolchenko, Z. Wan, and S. Arnold, “Taking whispering gallery-mode single virus detection and sizing to the limit,” Appl. Phys. Lett. 101(4), 043704 (2012).
], we will talk of shifts, splittings and linewidth changes in wavelength terms. Adsorption on the silica equator is calculated to produce a resonance shift of 0.25 fm, well below the 2fm r.m.s. noise in Ref.
6V. R. Dantham, S. Holler, V. Kolchenko, Z. Wan, and S. Arnold, “Taking whispering gallery-mode single virus detection and sizing to the limit,” Appl. Phys. Lett. 101(4), 043704 (2012).
. The calculated splitting would be 2x0.25 = 0.5 fm, much smaller than the experimental linewidth of 1950fm (
Q = 4x10
5). If 8 copies of the virus adsorbed randomly on the equator of this resonator the mean splitting for a statistical number of trials would be
, still much smaller than the line width. Even if we increase the Q to ~4x10
6 (line width = 195 fm), a value easily within reach of such a microcavity [
27W. Kim, Ş. K. Özdemir, J. Zhu, and L. Yang, “Observation and characterization of mode splitting in microsphere resonators in aquatic environment,” Appl. Phys. Lett. 98(14), 141106 (2011).
], the splitting due to these 8 viruses would still not be seen. It is to be noted that the mode splitting due to the imperfections (Rayleigh scattering) will be present when the ring resonator has an extremely high Q-factor (~10
8). With a low Q-factor, such a splitting does not occur. In our case, because of the lossy metallic nanoparticles, our Q-factor is much lower (~10
6).
Fig. 6 Comparison between the mode splitting magnitude upon adsorbing 8 randomly located viruses to a bare microsphere equator (black) with the additional splitting for binding the same viruses to periodic plasmonic nanoshell epitopes (N = 4) on a similar microsphere (red).
Next we consider the effect of including functionalized periodic plasmonic epitopes as binding sites. We suppose now that epitopes are added to the microcavity described in the preceding paragraph. These epitopes will be of the same type as the single epitope used in Ref.
6V. R. Dantham, S. Holler, V. Kolchenko, Z. Wan, and S. Arnold, “Taking whispering gallery-mode single virus detection and sizing to the limit,” Appl. Phys. Lett. 101(4), 043704 (2012).
(gold nanoshell with silica core). We will need some additional experimental information concerning the shift |
g| and linewidth change
produced by the addition of the single nanoshell; in wavelength terms we have measured these to be 175 fm and 230 fm, respectively, during our experiments described in Ref.
6V. R. Dantham, S. Holler, V. Kolchenko, Z. Wan, and S. Arnold, “Taking whispering gallery-mode single virus detection and sizing to the limit,” Appl. Phys. Lett. 101(4), 043704 (2012).
. From the inequality
Eq. (27) no splitting would be seen for the adsorption of a single nanoshell for a Q ~4x10
6 ; 2|g| = 350 fm does not exceed the overall linewidth of 425 fm (195 fm from the original linewidth plus 230 fm caused by the deposition of a nanoparticle). Splitting would be seen for two epitopes separated by an optical phase of
nπ since the magnitude of the splitting is 700 fm compared with an overall linewidth of 635 fm. With 4 optimally placed epitopes the splitting magnitude would be 2x4x175 = 1400 fm, well in excess of the overall linewidth. The key question now is: how much additional splitting will be produced by the binding of viruses onto the functionalized hotspots of these epitopes? From Ref.
6V. R. Dantham, S. Holler, V. Kolchenko, Z. Wan, and S. Arnold, “Taking whispering gallery-mode single virus detection and sizing to the limit,” Appl. Phys. Lett. 101(4), 043704 (2012).
, which only used one epitope, the maximum shift for a virus was 17 fm, which would have produced an unseen increase in splitting of 34 fm. With 8 viruses adsorbing on our 4 epitopes for which a splitting should be apparent, an increase in splitting of ~280 fm is calculated (upper trace in
Fig. 6). So we have gone from an invisible splitting of 3.5 fm by random adsorption on silica to a visible increase of 280 fm for adsorption onto a phased array of 4 epitopes.