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# 1 Summary of events: A vortex half ring is injected into vortex-free superflow, generated with rotation around the vertical axis (Ω=0.21rad/s or Reₛ≈30). The figures represent snapshots from simulation calculations in the rotating frame, which are consistent with the experimental observations. The only temperature dependence is contained in the mutual-friction parameters 𝛼 and 𝛼' which we take from Ref. []. (Top) At high temperatures the loop grows monotonically into a single rectilinear vortex line, pulled by the Magnus force from the superflow. The motion is highly damped since the Magnus force is balanced by the mutual friction force. (Bottom) At low temperatures the loop moves with the circulating superflow: (44 s) The vortex is unstable with respect to loop formation and develops through reconnections into a tangle at the injection site. (68 s) The Magnus force from the vortex-free superflow extends the tangle along the vertical axis. The measured flight time 𝜏_𝐹 for the tangle to extend across a vertical distance 𝑑 is controlled by mutual-friction damping: 𝜏_𝐹≈𝑑/(Ω𝑅𝛼). (108 — 160 s) The final step is the reconnection of the tangle to rectilinear lines. The growth in the relative polarization is experimentally observed as a relaxation in the NMR absorption and the total number of vortex lines in the final stable state is determined from the NMR line shape (Fig. ).

Transition to superfluid turbulence governed by an intrinsic parameter

A.P. Finne, T. Araki§, R. Blaauwgeers∗,†, V.B. Eltsov∗,‡, N.B. Kopnin∗,∥, M. Krusius, L. Skrbek1, M. Tsubota§, & G.E. Volovik∗,∥

Low Temperature Laboratory, Helsinki University of Technology, P.O.Box 2200, FIN-02015 HUT, Finland.
§ Department of Physics, Osaka City University, Sumiyoshi-Ku, Osaka 558-8585, Japan.
Kamerlingh Onnes Laboratory, Leiden University, P.O.Box 9504, 2300 RA Leiden, The Netherlands.
Kapitza Institute for Physical Problems, Kosygina 2, 119334 Moscow, Russia.
Landau Institute for Theoretical Physics, Kosygina 2, 119334 Moscow, Russia.
1 Joint Low Temperature Laboratory, Institute of Physics ASCR and Charles University, Prague, Czech Republic.

Hydrodynamic flow in both classical and quantum fluids can be either laminar or turbulent. To describe the latter, vortices in turbulent flow are modelled with stable vortex filaments. While this is an idealization in classical fluids, vortices are real topologically stable quantized objects in superfluids. Thus superfluid turbulence  is thought to hold the key to new understanding on turbulence in general. The fermion superfluid 3He offers further possibilities owing to a large variation in its hydrodynamic characteristics over the experimentally accessible temperatures. While studying the hydrodynamics of the B phase of superfluid 3He, we discovered a sharp transition at 0.60 Tc between two regimes, with regular behaviour at high-temperatures and turbulence at low-temperatures. Unlike in classical fluids, this transition is insensitive to velocity and occurs at a temperature where the dissipative vortex damping drops below a critical limit. This discovery resolves the conflict between existing high- and low-temperature measurements in 3He-B: At high temperatures in rotating flow a vortex loop injected into superflow has been observed to expand monotonically to a single rectilinear vortex line , while at very low temperatures a tangled network of quantized vortex lines can be generated in a quiescent bath with a vibrating wire . The solution of this conflict reveals a new intrinsic criterion for the existence of superfluid turbulence.

In conventional liquids the vorticity $`𝝎=\mathbf{}\times 𝐯`$ obeys the Navier-Stokes equation

$`{\displaystyle \frac{𝝎}{t}}=\mathbf{}\times [𝐯\times 𝝎]+\nu \mathbf{}^2𝝎.`$The interplay of the two terms on the rhs, the inertial first term and the viscous second term, governs the transition to turbulence . It is determined by the external conditions through the Reynolds number, $`Re=RU/\nu `$, formed by the characteristic size $`R`$ of the system, the flow velocity $`U`$, and the kinematic viscosity $`\nu `$. At large $`Re1`$, the effect of the inertial term is dominating, and laminar flow becomes increasingly unstable. If vorticity is released into a meta-stable laminar state, a sudden transition to a chaotic flow of eddies occurs. The evolution of the turbulent flow is described by the Kolmogorov energy cascade: the kinetic energy of the flow is transferred to ever smaller length scales, with large vortex loops decaying into smaller loops, until a scale is reached where the energy is dissipated by viscosity.

In superfluids turbulence acquires new features: A superfluid can be described as consisting of two inter-penetrating fractions, the frictionless (superfluid) and viscous (normal) components. If both fractions are moving, as is the case eg. behind a pulled grid in measurements on superfluid turbulence in 4He , the turbulent state bears more resemblance to that of viscous liquids. A new class of superfluid turbulence becomes possible when the normal component is so viscous that it is essentially immobile and only the superfluid component is moving with respect to the boundaries. This is the usual case in the flow of 3He, which is considered here.

The vorticity of the superfluid component is quantized in terms of the circulation quantum $`\kappa =2\pi \mathrm{}/M`$ where $`M`$ is the mass of a superfluid particle. The flow of the superfluid component can be characterized with the ‘superfluid Reynolds number’ $`Re_\mathrm{s}=RU/\kappa `$. The Feynman criterion $`Re_\mathrm{s}1`$ gives the velocity at which it becomes energetically favorable to form a quantized vortex. If a large nucleation barrier exists, vortices are not created and the superfluid remains vortex-free, in our measurements here up to $`Re_\mathrm{s}200`$. Quantized vortices can then be injected, either by some extrinsic means or by increasing the velocity so that the nucleation barrier is overcome and spontaneous vortex formation occurs. The limit of large $`Re_\mathrm{s}1`$ is equivalent to a vanishing Planck’s constant, $`\kappa \mathrm{}0`$, so that the vorticity becomes a continuous variable, like in the classical case. However, the fluid still remains unconventional because of its two-fluid nature: In addition to the superflow, there is the normal component which is at rest in the container frame, $`𝐯_\mathrm{n}=0`$. Interactions between the superfluid vorticity and the normal component give rise to a mutual friction force on a unit volume of the superfluid,

$`𝐟_{\mathrm{mf}}=\alpha ^{}\rho _\mathrm{s}[𝐯_\mathrm{s}\times 𝝎_\mathrm{s}]+\alpha \rho _\mathrm{s}[\widehat{𝝎}_\mathrm{s}\times [𝝎_\mathrm{s}\times 𝐯_s]],`$which is described by the dimensionless temperature-dependent parameters $`\alpha `$ and $`\alpha ^{}`$. These correspond to the usual mutual friction parameters, if we are dealing with one vortex or locally polarized vorticity, or they may be renormalized in the general case. Here $`𝐯_\mathrm{s}`$ and $`𝝎_\mathrm{s}=\mathbf{}\times 𝐯_\mathrm{s}`$ are the superfluid velocity and vorticity, and $`\widehat{𝝎}_\mathrm{s}`$ is the unit vector in the direction of $`𝝎_\mathrm{s}`$. Inserting the mutual-friction force in the Euler equation one obtains the hydrodynamic equation for $`Re_\mathrm{s}1`$ :

$$\frac{𝝎_\mathrm{s}}{t}=(1\alpha ^{})\mathbf{}\times [𝐯_\mathrm{s}\times 𝝎_\mathrm{s}]+\alpha \mathbf{}\times [\widehat{𝝎}_\mathrm{s}\times (𝝎_\mathrm{s}\times 𝐯_\mathrm{s})].$$
(1)
The energy dissipation is determined by the mutual friction damping $`\alpha `$ in the second term, while the reactive mutual-friction $`\alpha ^{}`$ renormalizes the inertial term of conventional hydrodynamics. Like in a classical fluid, the inertial term drives the instability by increasing the number of vortex loops, whereas the dissipative term acts to stabilize laminar flow by decreasing the number of loops. The fundamental difference from conventional hydrodynamics is that these two competing terms now have a similar dependence on the velocity and its gradients. Their relative importance is determined by the intrinsic dimensionless parameter of the superfluid, $`q=\alpha /(1\alpha ^{})`$, in contrast to classical liquids where it is governed via the Reynolds number by the extrinsic quantities $`R`$ and $`U`$. We thus expect laminar flow to become unstable when $`q`$ drops below a critical value $`q_c1`$. In the case of a single vortex line the parameter $`q`$ also marks the crossover from propagating Kelvin waves $`(q1)`$ to the over-damped regime $`(q1)`$ . In Fermi superfluids and superconductors with finite energy gap one has $`q(\omega _0\tau )^1`$, where $`\omega _0`$ is the spacing between the bound states of quasiparticles in the vortex core and $`\tau ^1`$ is their lifetime broadening, owing to scattering from the normal component . In 3He-B, $`q`$ approaches infinity at $`T_\mathrm{c}`$ and drops monotonically to zero, when cooled down to $`T0`$ . Such an evolution of $`q`$ with temperature would in classical fluids correspond to scanning the Reynolds number from 0 to $`\mathrm{}`$: We can thus study the emergence of turbulence as a function of temperature when $`q`$ falls below unity.

Measurements:—In rotating 3He-B high-velocity vortex-free superflow can be achieved, into which vortex seed loops can be injected (Fig. 1). We record as a function of temperature $`T`$ and rotation $`\mathrm{\Omega }`$ the number of rectilinear vortex lines after injection $`(N_\mathrm{f})`$, when all transients have died out. Non-invasive NMR measurement is used to determine the number of vortex lines in the final state. The NMR absorption spectrum of 3He-B maps the order-parameter texture, which is strongly affected by the orienting effect from the $`𝐯_\mathrm{s}`$ field. It allows us to determine $`N_\mathrm{f}`$ with a resolution better than 10 vortex lines in the temperature range of Fig. 2. In Fig. 2d we list the result after each injection event in the $`(\mathrm{\Omega },T)`$ plane, classified as a regular high-temperature (blue) or turbulent low-temperature (red) process. Here the injection method is the shear-flow instability of the phase boundary between the 3He-A and 3He-B phases, which produces a small random number ($`\mathrm{\Delta }N10`$) of B-phase vortex loops whose one end sticks out of the AB interface while the other end is on the cylindrical sample boundary . At high temperatures an injected loop expands and becomes a single rectilinear vortex line. At low temperatures the loop evolves into tangled vorticity which finally fills the whole sample volume with rectilinear lines. The central plot Fig. 2d leads to a surprising conclusion: Only a narrow cross-over region at $`0.60T_\mathrm{c}`$ separates the two processes, with little dependence on the maximum superflow velocity $`U=\mathrm{\Omega }R`$ of the initial vortex-free state. At the transition temperature $`0.60T_\mathrm{c}`$, the experimental value of $`q=\alpha /(1\alpha ^{})1.3`$ .

Vortices can also be injected with other methods. In the absence of the AB interface vortices are created (i) at a rough spot on the cylindrical sample boundary , (ii) under neutron irradiation , or (iii) by a sudden leak of vortex loops through the small orifice which connects the sample to the rest of the liquid 3He volume . These three processes have different critical velocities and temperature dependences, but in all cases the measurements are consistent with a crossover at $`0.60T_\mathrm{c}`$ from a low-yield to a high-yield process with decreasing temperature. Depending on sample geometry, below $`0.50T_\mathrm{c}`$ the vortex leak through the orifice may occur at 0.5 rad/s or less. Even at these low velocities $`(Re_\mathrm{s}70)`$ vortex loop expansion is found to be turbulent.

The long cylindrical sample is monitored with two signal coils at different locations, connected to independently operated NMR spectrometers. In addition to the number of vortex lines in the final stable state, we can obtain information on the transient state: (i) on the axial expansion of the vorticity and (ii) below $`0.60T_\mathrm{c}`$ on the decay of the tangle into a polarized array of rectilinear lines. The former process, the expansion of the vorticity along the $`\widehat{𝛀}`$ axis between the two signal coils, is controlled by the mutual-friction damping $`\alpha `$: The extracted flight time for the motion in the axial direction reproduces $`\alpha (T)`$ from Ref.  and is continuous across the transition at $`0.60T_\mathrm{c}`$. Thus the axial expansion is indifferent to whether the vorticity moves as noninteracting loops or as a network of loops.

Below $`0.60T_\mathrm{c}`$, the arrival of the tangled vorticity within a pick-up coil is signalled by a massive abrupt shift of the absorption close to the Larmor edge, where a new sharp absorption peak is formed (Fig. 2c). This line shape is similar to that measured in the non-rotating state $`(\mathrm{\Omega }=0)`$. Here the orientational effects from the superflow fields trapped around the vortex cores are averaged out, owing to the randomly distributed vortex tangle with density $`\omega _\mathrm{s}\omega _{\mathrm{eq}}`$ and the removal of essentially all large-scale vortex-free superflow. The sharp peak then decays with a relaxation time $`30`$s towards the broad final stable state spectrum of a polarized vortex array with rectilinear lines and $`\omega _\mathrm{s}=\omega _{\mathrm{eq}}=2\mathrm{\Omega }`$. This restructuring of the NMR line shape is thus a direct signal from the transient epoch with a vortex tangle.

Numerical simulation—As seen in Fig. 1, the measured features are consistent with our simulation calculations. We use the vortex filament model  in the rotating frame . A vortex is represented in parametric form by $`𝐬=𝐬(\xi ,t)`$, where $`𝐬`$ refers to a point on the filament and $`\xi `$ is the arc length along it. The spatial and time evolutions are integrated rigorously using the Biot-Savart law. The vortex velocity $`\dot{𝐬}`$ is calculated from the dynamic equation $`\dot{𝐬}=𝐯_{\mathrm{sl}}+\alpha ^{}𝐬^{}\times [𝐬^{}\times 𝐯_{\mathrm{sl}}]\alpha [𝐬^{}\times 𝐯_{\mathrm{sl}}]`$ (recall that $`𝐯_\mathrm{n}=0`$), where the local superfluid velocity $`𝐯_{\mathrm{sl}}`$ includes all contributions to the superflow at $`𝐬(\xi ,t)`$. As boundary conditions we use smooth solid-walls with image vortices.

The initial state is a vortex half ring (diameter 3.5 mm) in the centre of a cubic sample container (10 mm wide). At high temperatures $`T/T_\mathrm{c}=0.8`$ the high damping $`\alpha `$ causes the seed loop to expand, mostly by motion in the radial and axial directions, into a single rectilinear vortex line in the centre of the sample. At low temperatures $`T/T_\mathrm{c}=0.4`$ the seed loop travels azimuthally and undergoes the Kelvin-wave instability, when the superflow is oriented along the vortex . Here $`q<1`$ so that Kelvin wave oscillations are only lightly damped and loop formation starts via reconnections. Thus a vortex network develops already at the injection site. The vortex-free superflow above the tangle stretches the loops with right orientation, while others shrink, and thus the tangle extends and travels in the axial direction. Through reconnections the properly oriented loops are gradually joined to lines which form a bundle almost parallel to the rotation axis $`\widehat{𝛀}`$. The bundle precesses initially around the $`\widehat{𝛀}`$ axis, but presumably more reconnections of remaining loops will continue adding lines to the bundle and thereby reducing the superflow around the bundle, until the equilibrium number of rectilinear vortex lines is reached. The complete calculation is so far too time consuming.

Discussion:—Both the results from the numerical simulations in Fig. 1 and from the measurements in Fig. 2 support the conclusion from Eq. (1) that the transition to turbulence is determined by the parameter $`q=\alpha /(1\alpha ^{})`$: There exists a critical value $`q_\mathrm{c}1`$ which separates the mutual-friction-dominated regime at $`q>q_\mathrm{c}`$ from the inertia-dominated turbulent regime at $`q<q_\mathrm{c}`$. With $`q>q_\mathrm{c}`$, the vortex-loop expansion is a regular stable process even in high-velocity superflow with $`Re_\mathrm{s}1`$, as has been verified in many previous measurements. With $`q<q_\mathrm{c}`$, a multiplication process is switched on, the vortex lines create loops, become entangled, and reconnect forming new loops. Could $`q_\mathrm{c}`$ be a fundamental universal number which, when $`Re_\mathrm{s}1`$, places the upper bound to superfluid turbulence?

Why has such a transition, driven by an intrinsic parameter, not been observed previously in superfluid 4He? A number of reasons can be listed: (i) In 3He the viscosity of the normal fraction is four orders of magnitude higher. Thus there is no ambiguity about the state of the normal component, unlike in the case of 4He. (ii) The vortex-core radius in 3He-B exceeds by two orders of magnitude that of 4He, where it is only of atomic size. Therefore vortex pinning and remanence at solid walls can often be neglected in 3He-B and metastable states with rapid superflow become possible. (iii) The most important difference lies in the value of mutual friction. In 4He the dissipative part $`\alpha `$ is small and influences vortex motion relatively little. Only within a narrow temperature interval $`\mathrm{\Delta }T/T_\lambda 210^3`$ from $`T_\lambda `$ one finds $`q>1`$ and expects different behavior. No conclusive measurements exist from so close to $`T_\lambda `$.

In the helium superfluids, the transition at $`q_\mathrm{c}1`$ lies very close to $`T_\lambda `$ in 4He, in the middle of the experimentally accessible temperature range in 3He-B, and below that at extremely low temperatures in 3He-A. This prediction is consistent with present experimental information. Here we found that turbulence is unstable even at high $`Re_\mathrm{s}`$, if $`q>q_\mathrm{c}`$. Perhaps in other multi-component hydrodynamic systems, superfluid or viscous, the stability of turbulence might also be governed by an intrinsic parameter, in addition to the Reynolds number.

This work was supported in part by the following EU and ESF programmes: EU-IHP ULTI-3, ESF-COSLAB, ESF-VORTEX. NK and GV are grateful to the Russian Foundation for Basic Research; LS thanks for the grant GAČR (202/02/0251). We thank W.F. Vinen for valuable discussion.


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# Aharonov-Bohm signature for neutral excitons in type-II quantum dot ensembles

## Abstract

The Aharonov-Bohm (AB) effect is commonly believed to be a typical feature of the motion of a charged particle interacting with the electromagnetic vector potential. Here we present a magnetophotoluminescence study of type-II InP/GaAs self-assembled quantum dots, revealing the Aharonov-Bohm-type oscillations for neutral excitons when the hole ground state changes its angular momentum from $`\mathrm{}_h`$ = 0 to $`\mathrm{}_h`$ = 1, 2, and 3. The hole ring parameters derived from a simple model are in excellent agreement with the structural parameters for this system.

In 1959, Aharonov and Bohm proposed an experiment to assess the manifestations of the electromagnetic potentials in the quantum domain aharonov59 . By allowing a charged particle to circulate a confined magnetic field flux region, they showed that surprisingly, there exist effects of the vector potential on the charged particles moving outside. After the circulation the particle wavefunction acquires a phase that is proportional to the magnetic flux contained within the closed path berry84 . Therefore, all observable phenomena depend upon the flux $`\mathrm{\Phi }`$ through the excluded region, and are shown to be periodic with period $`\mathrm{\Phi }_0=hc/e`$ byers61 ; bloch68 ; buttiker83 . This oscillatory characteristic is the signature of the Aharonov-Bohm (AB) effect aharonov59 ; peshkin89 .

The definite measurement of the original AB effect was performed by Tonomura et al. in a very clever experiment tonomura86 ; peshkin89 . Investigation and applications of the AB concepts in condensed matter have been carried out in metals webb85  and superconductors byers61 ; bloch68 ; parks64 . With the advance of lithography and semiconductor growth techniques, it was possible to devise mesoscopic timp87 ; ford88  and nanoscopic fuhrer01 ; yau02  devices where many theoretically predicted signatures evidencing ring-like electronic properties altshuler81 ; buttiker85 ; aronov87  have been observed by transport experiments. For the case of few electrons, self-assembled quantum rings lorke00  were used for the observation of the characteristic spectra including excitations of states of different angular momentum lorke00 . Due to the small sizes of these structures, it was possible for the carriers to maintain the coherence of their wavefunctions during their motion (at sufficiently low temperatures), thus allowing the observation of relative quantum phase effects (AB-like).

It was proposed that a neutral excitation could also exhibit such an AB oscillatory behavior chaplik95 ; kalameitsev98 . Electron-hole pairs created by optical excitation and bound together via Coulomb interaction (excitons) are a good candidate when either one or both carriers are confined to a ring-like geometry. The existence of the AB oscillations for neutral excitons in semiconductor quantum rings, where both carriers are confined in the rings, has been a matter of controversy in recent years chaplik95 ; romer00 ; song01 ; hu01 ; govorov02a ; govorov02b  since some of the work predicted that the AB oscillations might be very weak because of exponentially small electron-to-hole tunnel amplitude chaplik95 ; kalameitsev98  or would vanish for a finite width ring song01 ; hu01 . Recently, the AB effect was observed in optical experiments by Bayer et al. on quantum-rings fabricated by lithographical methods bayer03 . They recorded AB oscillations for charged excitons. Perhaps due to the aforementioned reasons, the AB effect was not found in the spectra of neutral excitons. The second possibility for observing AB-like oscillations in the optical spectra of a neutral excitation is the case of type-II quantum dots (QD) kalameitsev98 ; govorov02a ; janssens01 ; janssens02 , where the confinement of one carrier inside the QD and the other carrier in the barrier naturally creates a ring-like structure (see Fig. 1). In such a structure, the carrier outside the QD would be bound to that confined in the QD via Coulomb attraction. This spatial charge separation produces a polarization of the exciton and thus AB oscillations could be observed in the energy of those carriers that are confined in the ring-shaped orbit around the QD. Moreover, the AB effect with a polarized exciton may not involve an electron-to-hole tunnel amplitude kalameitsev98 ; govorov02a . There are several self-assembled quantum dot systems where a type-II heterojunction is expected; for instance, GaSb on GaAs hatami95  and Ge on Si schittenhelm95  provide a three-dimensional confinement for the holes, thereby creating a ring-like potential for the electrons. Alternatively, InP on InGaP hayne00  and InP on GaAs ribeiro02  provide the localization of the electrons in the QD, and consequently a ring-like hole wavefunction. Figure 1 sketches the valence band potential in a very simplified way, not taking into account the contribution of the strain. It is known that the QD strains the GaAs substrate and cap layer, and that in turn changes the potential landscape in the GaAs, creating an additional ring-like potential around the QD for heavy-hole confinement xu91 . The light hole may have a well-like strain-induced potential below the QD xu91 , however being shallower than the heavy hole ring potential. InP QDs are particularly suitable for observing oscillatory behavior with AB period $`\mathrm{\Phi }_0`$. With diameters of the order of 35 nm one obtains a period of few Tesla for the AB effect.

Here we present data on InP/GaAs self-assembled quantum dot ensembles where the confined magnetoexcitons trap fluxes in the range of 0-3 flux quanta, corresponding to a magnetic field span of 0-12 T. Samples were grown at different rates and PH3 fluxes by metal-organic chemical vapor deposition in a commercial reactor on nominally flat semi-insulating Cr-doped GaAs  substrates at 550 oC ribeiro02 . The structures consisted of a 300 nm undoped GaAs grown at 600 oC buffer layer followed by the QD layer. The islands were then capped with a 50 nm undoped GaAs cap. For this work, the sample with the narrowest luminescence line width ($``$35 meV) was chosen, representing an ensemble of $`3\times 10^{10}`$ QDs/cm2 ribeiro02  with small size dispersion. The dimensions of the QDs were determined by cross-section transmission electron microscopy (TEM) maltez03 , and were found to be (32 $`\pm `$ 6) nm and (4 $`\pm `$ 2) nm in average diameter and height. The InP QDs are not cylindrically symmetric, being elongated in the  direction. The photoluminescence (PL) experiments were performed at 2 K, with magnetic fields up to 12 T (in 0.2 T steps), and using an Ar laser with 2 W/cm2 as excitation source. These conditions guaranteed the filling of only the system ground state, allowing the assessment of single particle energies. For higher excitation power a line broadening and luminescence shift nakaema02  takes place, indicating excited state population, which would complicate our analysis.

Fig. 2 shows the InP QDs PL spectra for $`B=0`$ (circles) and $`B=12`$ T (triangles). The total energy shift within this field range was 4.5 meV. The PL peak intensity increased about 50% from 0 to 12 T, indicating an enhancement of the electron and hole wavefunction overlap. The spectra for all the field values were fit with a single Gaussian line shape in order to extract the evolution of the PL peak position of the InP QDs as a function of the applied magnetic field, $`E_{PL}(B)`$, which is plotted in Fig. 3(a). There one can see that instead of the typical monotonic diamagnetic energy shift characteristics of type-I QD wang96 , the InP QD PL shows an oscillatory behavior in the $`E_{PL}(B)`$ curve, very similar to what has been predicted by Kalameǐtsev and collaborators kalameitsev98 . In that work, it was expected that the maxima of oscillating part in $`E_{PL}(B)`$ would develop every time that the lower-lying carrier state changed its angular momentum from $`\mathrm{}_e`$ = 0 to $`\mathrm{}_e`$ = $`1`$ and so forth. These transitions were seen to occur when $`R^2/l_B^2|\mathrm{}_e|`$ kalameitsev98 , with $`R`$ being the ring radius, $`l_B^2=\mathrm{}/eB`$ the magnetic length and $`\mathrm{}_e`$ the electron state angular momentum. Rewriting this expression for the holes in InP QDs, $`R(\mathrm{}\mathrm{}_h/eB)^{1/2}`$ and then by inserting the experimental values of $`B`$ for the observed plateau-like regions \[Figure 3(a)\] an estimate for the hole ring radius can be obtained. Averaging the results for the three plateaux, a radius of $`R=(16\pm 1)`$ nm is obtained, in agreement to what has been measured by TEM maltez03 .

Nevertheless, it is possible to further elaborate the analysis of the data. The electron ground state of InP/GaAs QDs has been investigated by capacitance spectroscopy (CV) bufon04 . It has already been established wang96  that the lateral confinement for the self-assembled quantum dot can be reasonably modelled by a parabolic potential. The diamagnetic shift of the electron ground state can be described by $`E_e=((\mathrm{}\omega _0)^2+(\mathrm{}\omega _c)^2/4)^{1/2}`$ fock28 , with $`\omega _c`$ being the cyclotron frequency. For InP/GaAs QDs grown under identical nominal conditions, a characteristic energy of $`\mathrm{}\omega _0=(5.6\pm 0.9)`$ meV was inferred bufon04 . Bearing this information in mind, an expression for $`E_{PL}(B)`$ can be written, consisting of three terms: a constant contribution, related to the energy gap of the material and the exciton binding energy; the diamagnetic energy shift for the electrons, discussed above; and the variation of the hole energy with the magnetic field govorov02b ; janssens01 :

$$E_{PL}(R,B)=E_g+\sqrt{(\mathrm{}\omega _0)^2+\left(\frac{\mathrm{}\omega _c}{2}\right)^2}+\frac{\mathrm{}^2}{2m_h^{}R^2}\left(\mathrm{}_h\frac{\mathrm{\Phi }}{\mathrm{\Phi }_0}\right)^2,$$
(1)
where $`m_h^{}`$ is the hole effective mass, $`\mathrm{\Phi }_0=hc/e`$ is the flux quantum and $`\mathrm{\Phi }=\pi R^2B`$ is the total magnetic flux through the ring of radius $`R`$. Using the above result for $`\mathrm{}\omega _0`$, both the constant and the electron contributions in the experimental data \[see solid curve in Fig. 3(a)\] can be subtracted. The result is shown in Fig. 3(b), where:

$$E_h(R,B)=E_{PL}(R,B)E_gE_e=\frac{\mathrm{}^2}{2m_h^{}R^2}\left(\mathrm{}_h\frac{\mathrm{\Phi }}{\mathrm{\Phi }_0}\right)^2.$$
(2)
As shown in Fig 3(b), $`E_h`$ (open squares) clearly oscillates as a function of the magnetic field, indicating phase coherence for the hole wavefunctions. Since $`\mathrm{\Phi }_0`$ is a constant (the Aharonov-Bohm period of the excitonic energy oscillations aharonov59 ; buttiker85 ; peshkin89 ), the experimental data were fit with the parabolas given by Eq. (2), one for each value of $`\mathrm{}_h`$ = 0, 1, 2, and 3, indicating the number of flux quanta enclosed by the hole ring. The result is shown by the dashed curves in the upper part of Fig. 3(b), from which $`R=(19.1\pm 0.4)`$ nm and $`\mathrm{\Phi }_0`$ = 3.61 T were obtained. One should keep in mind that the hole ring must have a radius slightly larger than the QD diameter (Fig. 1), and therefore the obtained values are in excellent agreement with the structural and electronic results discussed above maltez03 .

It is worth noting that an ensemble of islands and not a single QD was probed. Nevertheless, the size dispersion did not suppress the AB oscillations. Some considerations can be done on the influence of the size dispersion $`\mathrm{\Delta }R`$ on the experimental results. From CV experiments bufon04  an effective size distribution of $`\mathrm{\Delta }R_{CV}`$ = 0.8 nm was inferred for the electron-confining potential. This CV estimate for $`\mathrm{\Delta }R`$ would be a lower bound for the experimentally-probed size dispersion. In order to model in a more quantitative fashion the effect of the QD ensemble on the hole-ring energy oscillations, $`E_h(R,B)`$ \[eq. (2)\] was calculated and weighted by a Gaussian size distribution function, where $`R`$ determined in Fig. 3(b) is the central peak radius and $`2\mathrm{\Delta }R_{CV}`$ is its full width at half maximum. The result is plotted in Fig. 3(b) (solid curve). It can be seen that the main effect of the size dispersion on $`E_h`$ is a progressive attenuation of the oscillation intensity, as one would expect, and it is consistent with the decrease in the amplitude of the data in Fig. 3(b). By comparing the data and the solid curve in Fig. 3(b) one can see that the experimental data of the hole ring are consistent with an effective size dispersion obtained from $`\mathrm{\Delta }R_{CV}`$.

Although describing the oscillation period and damping in a good agreement with independent structural/electronic results, this simplified model fails to predict the energy offset of the last parabola (corresponding to $`\mathrm{}=3`$). The contributions for the offset might be: (i) the experimental uncertainty in $`\mathrm{}\omega _0`$ obtained from the CV fit \[solid curve in Fig 3(a)\]; (ii) a possible dependence of the exciton binding energy on the magnetic field; (iii) deviations from the parabolic lateral confinement due to the actual QD morphology. Clearly, a more detailed and complete modelling of the system, including the points indicated above, is needed for understanding all the experimental features presented in Fig. 3(b), but such a task is beyond the scope of the present work. Nevertheless, the simple model used here consistently describes the main physical issues connected to the observation of the hole AB oscillations (Fig. 3).

In summary, we have presented clear evidence of Aharonov-Bohm oscillations in the spectrum of electrically neutral excitons confined in type-II InP/GaAs quantum dots. We have observed the trapping of up to three flux quanta inside the ring trajectory of the hole, for a reasonable span of magnetic fields. Despite the presence of a size dispersion, the AB oscillations were easily observable, evidencing the phase coherence for holes moving around QDs.

###### Acknowledgements.

The magnetic field experiments were performed at the Optical Properties Group facilities at Campinas State University (Unicamp). We would like to thank H. Gazetta Filho for the technical support on sample growth. We gratefully acknowledge financial support from Fundação de Amparo à Pesquisa do Estado de São Paulo and MCT-CNPq.


text:
# Onset of DNA Aggregation in Presence of Monovalent and Multivalent Counterions

## INTRODUCTION

Condensation and aggregation of DNA, induced by multivalent counterions, have been extensively studied in the past two decades (for a review, see Bloomfield et al. (2000) and references therein). The term condensation usually refers to the collapse of a single, long DNA chain. Condensation plays an important role in storage and packing of DNA; for example, in viral capsids Gelbart et al. (2000). Aggregation of DNA is a closely related phenomenon, where multiple chains attract each other and form a variety of condensed mesophases of complex structure Pelta et al. (1996b, a). In both phenomena multivalent counterions play a crucial role, screening the electrostatic repulsion between charged strands of DNA and mediating an effective attraction.

A variety of tri- and tetra-valent ions can induce aggregation and condensation, among them the polyamines spermidine ($`3^+`$) and spermine ($`4^+`$) Tabor and Tabor (1984); Gosule and Schellman (1978); Chattoraj et al. (1978), as well as cobalt-hexamine Widom and Baldwin (1980, 1983). In typical experiments on aggregation Pelta et al. (1996b); Raspaud et al. (1998); Saminathan et al. (1999) multivalent ions are gradually added to a solution with fixed concentration of DNA segments and monovalent salt. Two such examples for spermine and spermidine are reproduced in Fig. 1 Pelta et al. (1996b). As the multivalent ion concentration is raised above a certain threshold, DNA segments begin to aggregate, and precipitate from the solution. Above the aggregation threshold, the DNA concentration decreases gradually or abruptly, depending on various parameters such as the monovalent salt concentration and total DNA concentration. Further addition of multivalent ions at higher concentrations reverses the aggregation. Above a second, re-dissolution threshold, all the DNA is re-dissolved in the solution (Fig. 1). The re-dissolution threshold (above which all the DNA re-dissolves) is almost independent on the DNA concentration. Its value can be attributed to screening of electrostatic interactions by multivalent ions Raspaud et al. (1998).

The aggregation threshold, where the onset of aggregation occurs, is the main experimental phenomenon addressed in our theoretical paper. The multivalent ion concentration at the onset depends strongly on the monovalent salt and DNA concentrations. This dependence has been recently measured in detail for short (150 base pair) DNA segments in presence of spermine Raspaud et al. (1998), and is reproduced in Fig. 2. The figure shows measurements of spermine concentrations at the onset of aggregation, for DNA concentrations ranging over four orders of magnitude and for four different monovalent salt concentrations: $`2`$, $`13`$, $`23`$ and $`88\mathrm{mM}`$. At very low DNA concentration, the spermine concentration depends strongly on the monovalent salt concentration. At higher DNA concentration it has only a weak dependence on the monovalent ion concentration but the spermine concentration is proportional to the DNA concentration, indicating that a certain number of spermine counterions are required, per DNA base, in order to induce aggregation. The solid line in Fig. 2, adapted from Raspaud et al. (1998), corresponds to a ratio: $`c_{z,\mathrm{aggr}}/c_{\mathrm{DNA}}=0.20`$, where $`c_{z,\mathrm{aggr}}`$ is the spermine concentration at the aggregation onset and $`c_{\mathrm{DNA}}`$ is the DNA concentration. This linear relation fits a large number of the experimental points in the intermediate DNA concentration range. It has been suggested by Raspaud et al. (1998, 1999) that the deviations from this line, at low and high DNA concentrations, represent two distinct physical regimes that need to be analyzed separately from the intermediate regime, where the linear fit works well.

In this work we focus on the onset of aggregation, and specifically on its dependence on the DNA concentration. We show that this dependence is simple for all the range of DNA concentration. Furthermore, for $`c_{\mathrm{DNA}}`$ smaller than the monovalent salt concentration we show that this dependence is linear: $`c_{z,\mathrm{aggr}}=\alpha c_{\mathrm{DNA}}+\beta `$. The coefficient $`\beta `$ is the multivalent counterion concentration far away from the DNA chains, while $`\alpha `$ accounts for the excess of multivalent ions around each chain. These quantities can be extracted, e.g., from the four experimental curves of Fig. 2. Several further conclusions are then drawn on the onset of DNA aggregation and on the counterion distribution around each double-stranded DNA.

## THEORETICAL CONSIDERATIONS

Consider an aqueous solution containing monovalent (1:1) salt, multivalent ($`z`$:1) salt and DNA segments below their threshold for aggregation. Throughout this paper, the DNA solution is assumed to be dilute enough such that the DNA segments do not overlap. We also assume that these DNA segments can be regarded as rigid rods. The concentrations of added monovalent salt, multivalent salt and DNA monomers are denoted by $`c_s`$, $`c_z`$ and $`c_{\mathrm{DNA}}`$, respectively. These are the solute concentrations per unit volume as controlled and adjusted in experiments. We will assume that the monovalent and multivalent salts have the same type of co-ion, so that altogether there are three ion species in the solution:

1. A multivalent counterion contributed from the $`z`$:1 multivalent salt, of concentration $`c_z`$.
2. A monovalent counterion contributed by monovalent salt of concentration $`c_s`$, and by counterions dissociated from the DNA, of concentration $`c_{\mathrm{DNA}}`$: in total, $`c_s+c_{\mathrm{DNA}}`$.
3. Co-ions coming from both $`z`$:1 and 1:1 salts, of concentration $`c_s+zc_z`$.

Each DNA segment attracts a layer of oppositely charged counterions referred to as the condensed counterions. As long as the typical distance between segments is large compared to the electrostatic screening length $`\kappa ^1`$, the electrostatic potential decays exponentially to zero far away from the DNA segments. In turn, the concentrations of the three ion species decay to well defined bulk values denoted by $`c_1^{\mathrm{}}`$ for the monovalent ions and $`c_z^{\mathrm{}}`$ for the $`z`$-valent ones. These concentrations should be distinguished from the concentrations $`c_s`$ and $`c_z`$ introduced above, which are the average concentrations of added salts regulated experimentally.

The Debye screening length, $`\kappa ^1`$, characterizing the exponential decay of the electrostatic potential, is determined by the bulk concentrations of all three ionic species:

$$\kappa ^2=4\pi l_B\left[c_1^{\mathrm{}}+z^2c_z^{\mathrm{}}+(c_1^{\mathrm{}}+zc_z^{\mathrm{}})\right].$$
(1)
where the third term is the co-ion concentration. It is equal to $`c_1^{\mathrm{}}+zc_z^{\mathrm{}}`$ due to charge neutrality far from the DNA where the potential decays to zero. The above equation makes use of the Bjerrum length, $`l_B=e^2/(\epsilon k_BT)`$, equal to about $`7`$ Å in aqueous solution at room temperature, $`k_BT`$ is the thermal energy , $`e`$ is the electron charge and $`\epsilon =80`$ is the dielectric constant of water. The Debye length as well as $`c_z^{\mathrm{}}`$ are shown schematically in Fig. 3. Other quantities that will be defined below are also indicated in this figure.

In dilute solutions different DNA segments do not overlap. Following previous works, we introduce a cell model also shown schematically in Fig. 3. Note that the model serves to illustrate the subsequent derivations but is not essential for the validity of our main results. In the cell model, each segment, of a cylindrical cross-section, is at the center of a cylindrical cell of radius $`R`$ and area $`A=\pi R^2`$ such that

$$c_{\mathrm{DNA}}=1/(aA).$$
(2)
Namely, each DNA monomer occupies a specific volume $`aA`$, where $`a1.7`$Å is the average charge separation on the chain taken hereafter as the monomer length.

We will assume below that the DNA solution is dilute enough so that $`R`$ is large compared to the Debye length $`\kappa ^1`$. This assumption is essential for our derivation and can be verified for all the experimental data considered in this paper. Density profiles of the three ion species are then practically identical to those near an isolated DNA segment with the same bulk concentrations $`c_1^{\mathrm{}}`$, $`c_z^{\mathrm{}}`$. In other words, the profiles are determined uniquely by $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$, with practically no dependence (or, more precisely, an exponentially small dependence) on the DNA monomer concentration. A demonstration of this claim is presented in Fig. 4, using the Poisson-Boltzmann theory in a cell model. For two very different values of $`R`$ corresponding to different $`c_{\mathrm{DNA}}`$, the counterion profiles match perfectly when the values of $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$ are the same. Note that the average concentrations of added salts, $`c_s`$ and $`c_z`$, have different values in the two cells because of the contribution of condensed ions.

The total number of $`z`$-valent counterions, per cell unit length, is given by:

$$Ac_z=Ac_z^{\mathrm{}}+\rho _z(c_1^{\mathrm{}},c_z^{\mathrm{}})$$
(3)
where $`\rho _z`$ is the excess number of $`z`$-valent ions per unit length near the DNA. Throughout the paper we use the symbol $`c`$ to denote concentrations per unit volume and $`\rho `$ for concentrations per DNA unit length. The excess $`\rho _z`$ can be evaluated in the limit of infinite cell radius, corresponding to an isolated chain:

$$\rho _z=2\pi _0^{\mathrm{}}rdr\left[n_z(r)c_z^{\mathrm{}}\right],$$
(4)
where $`n_z(r)`$ is the $`z`$-valent local counterion concentration at distance $`r`$ from the axis of symmetry, and $`n_z(\mathrm{})=c_z^{\mathrm{}}`$. Following the discussion in the previous paragraph, the excess $`\rho _z`$ is determined uniquely by $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$. Its exact functional dependence on these variables is generally not known, although it can be evaluated approximately, e.g., using the Poisson-Boltzmann equation or in computer simulations.

For monovalent counterions we have, in a similar fashion:

$$Ac_s+Ac_{\mathrm{DNA}}=Ac_1^{\mathrm{}}+\rho _1(c_1^{\mathrm{}},c_z^{\mathrm{}}),$$
(5)
where $`\rho _1`$, the excess of monovalent counterions per unit length, is defined as in Eq. 4, and $`Ac_{\mathrm{DNA}}=1/a`$ is the DNA charge density per unit length. The extra term in the left-hand-side of Eq. 5 originates from monovalent counterions contributed by the DNA monomers. Using Eq. 2 we can rewrite Eqs. 3 and 5 as:

$$c_z=c_z^{\mathrm{}}+a\rho _z(c_1^{\mathrm{}},c_z^{\mathrm{}})c_{\mathrm{DNA}}$$
(6)
and

$$c_s=c_1^{\mathrm{}}+\left[a\rho _1(c_1^{\mathrm{}},c_z^{\mathrm{}})1\right]c_{\mathrm{DNA}}.$$
(7)
These two equations relate the experimentally adjustable $`c_s`$, $`c_z`$ and $`c_{\mathrm{DNA}}`$ to the bulk densities $`c_1^{\mathrm{}}`$, $`c_z^{\mathrm{}}`$ that in turn, are important because they determine the ion density profiles.

In the limit of infinite DNA dilution, $`c_{\mathrm{DNA}}=0`$, and therefore $`c_z=c_z^{\mathrm{}}`$ and $`c_s=c_1^{\mathrm{}}`$. At any finite DNA concentration $`c_z`$ and $`c_s`$ are not equal to $`c_z^{\mathrm{}}`$ and $`c_1^{\mathrm{}}`$, respectively, because each segment captures some of the multivalent ions and releases a number of monovalent ones. Equations 6 and 7 express the correction to $`c_s,c_z`$ at given $`c_1^{\mathrm{}},c_z^{\mathrm{}}`$ for both mono- and multi-valent counterion species. The dimensionless quantities $`a\rho _1,a\rho _z`$ are the excess of the mono- and multi-valent counterion species, respectively, per DNA monomer.

We would like to emphasize the generality of Eqs. 6 and 7. They do not depend on the assumption of parallel DNA residing in the middle of oriented cylindrical unit cells, or on any mean-field approximation for the distribution of counterions. The only assumption required to derive Eqs. 6 and 7 is that the average distance between DNA segments is large compared with the Debye length. Although Eqs. 6 and 7 are correct for any $`c_s`$, $`c_z`$ and $`c_{\mathrm{DNA}}`$ below the onset of DNA aggregation, we will be interested below specifically in the aggregation onset.

Onset of aggregation

Our aim now is to find how the value of $`c_z`$ at the onset of aggregation, $`c_{z,\mathrm{aggr}}`$, depends on $`c_{\mathrm{DNA}}`$. We will assume that this aggregation onset depends on $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$, but not on the average distance between DNA chains. We motivate this assumption by the fact that $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$ determine the density profile of multivalent counterions around the DNA chains, which, in turn, mediate the attraction necessary for aggregation. Before discussing this assumption in more detail, let us first consider its consequences. We can imagine an experiment where $`c_z^{\mathrm{}}`$ is gradually increased while $`c_1^{\mathrm{}}`$ is kept fixed. Aggregation will start, in this experiment, above a certain threshold value of $`c_z^{\mathrm{}}`$. Our assumption is that this threshold does not depend on $`c_{\mathrm{DNA}}`$. In real experiments, however, $`c_z`$ is adjusted rather than $`c_z^{\mathrm{}}`$, and $`c_s`$ is kept fixed rather than $`c_1^{\mathrm{}}`$. In order to find the threshold value in terms of the experimentally available $`c_z`$ we need to map $`c_1^{\mathrm{}},c_z^{\mathrm{}}`$ onto $`c_s,c_z`$. This mapping is described by Eqs. 67, and involves $`c_{\mathrm{DNA}}`$. It is only through this mapping that $`c_{\mathrm{DNA}}`$ will affect the threshold of aggregation.

The limit of $`c_{\mathrm{DNA}}c_s`$:

The limit $`c_{\mathrm{DNA}}c_s`$ offers a particularly simple dependence of $`c_{z,\mathrm{aggr}}`$ on $`c_{\mathrm{DNA}}`$ and is considered first. Most models and experiments indicate that monovalent counterions cannot overcharge DNA segments. Hence the monovalent excess, $`a\rho _1`$, in Eq. 7, is a number between zero and one, because the excess monovalent charge is smaller than that of DNA. From Eq. 7 $`|c_sc_1^{\mathrm{}}|c_s`$ as long as $`c_{\mathrm{DNA}}c_s`$. It is then possible to replace $`c_1^{\mathrm{}}`$ by $`c_s`$, leading to a simplification of Eq. 6:

$$c_z=c_z^{\mathrm{}}+a\rho _z(c_s,c_z^{\mathrm{}})c_{\mathrm{DNA}}.$$
(8)
Note that $`c_{\mathrm{DNA}}`$ is indeed smaller than $`c_s`$ in most of the experimental points in Fig. 2. However a similar simplification cannot be applied for $`c_z`$ because it is typically much smaller than $`c_s`$, and often smaller than $`c_{\mathrm{DNA}}`$.

According to our principal assumption, aggregation starts at a threshold value $`c_z^{\mathrm{}}=c_z^{}`$, which does not depend on $`c_{\mathrm{DNA}}`$ (while $`c_{z,\mathrm{aggr}}`$, the average multivalent salt concentration does depend on $`c_{\mathrm{DNA}}`$ through Eq. 8). Similarly, the density profile at the threshold does not depend on $`c_{\mathrm{DNA}}`$, because it is determined by $`c_1^{\mathrm{}}=c_s`$ and $`c_z^{}`$. The excess of $`z`$-valent counterions, as determined from this profile, is equal to:

$$\rho _z^{}=\rho _z(c_s,c_z^{}),$$
(9)
with no dependence on $`c_{\mathrm{DNA}}`$. Using the threshold values $`c_z^{}`$ and $`\rho _z^{}`$ in Eq. 8, we find that the average concentration of $`z`$-valent ions at the onset of aggregation is:

$$c_{z,\mathrm{aggr}}(c_{\mathrm{DNA}})=c_z^{}+a\rho _z^{}c_{\mathrm{DNA}}.$$
(10)
This is the threshold concentration that was measured experimentally in Raspaud et al. (1998). Note that in Eq. 10 $`c_z^{}`$ as well as $`\rho _z^{}`$ depend on the monovalent salt concentration, $`c_s`$, but the explicit dependence is omitted for clarity.

The simple relationship expressed by Eq. 10 is one of our main results. As a visualization of this result we refer again to Fig. 3. The quantities $`\rho _z`$, $`c_z^{\mathrm{}}`$ and the density profile $`n_z(r)`$ are indicated in this figure. At the onset of aggregation $`c_z^{\mathrm{}}`$ is equal to $`c_z^{}`$ and does not depend on $`c_{\mathrm{DNA}}`$ (or equivalently, on the spacing between DNA segments, $`R`$). As $`c_{\mathrm{DNA}}`$ is increased the distance between DNA strands decreases. The onset values of $`c_z^{\mathrm{}}`$ and $`\rho _z`$ do not change, but the contribution of $`\rho _z`$ to the average concentration increases, leading to an increase in $`c_{z,\mathrm{aggr}}`$.

The coefficients $`a\rho _z^{}`$ and $`c_z^{}`$ of the linear dependence in Eq. 10 are the coefficients $`\alpha `$ and $`\beta `$ defined in the introduction section. They can be easily found from the experimental data: $`c_z^{}`$ is the value of $`c_{z,\mathrm{aggr}}`$ in the limit of infinite DNA dilution, $`c_{\mathrm{DNA}}0`$, since in this limit $`c_z=c_z^{\mathrm{}}=c_z^{}`$. The excess at the onset, $`\rho _z^{}`$, can be found from the slope of $`c_{z,\mathrm{aggr}}`$ as function of $`c_{\mathrm{DNA}}`$. Before presenting a detailed comparison with experiments, we generalize the treatment for small $`c_{\mathrm{DNA}}`$ to arbitrary values.

The case of $`c_{\mathrm{DNA}}c_s`$ :

When $`c_{\mathrm{DNA}}`$ is of the same order as $`c_s`$ or larger, corrections to $`c_1^{\mathrm{}}`$ must be taken into account, as expressed by Eq. 7, and the linear relation of Eq. 10 no longer holds. The ion density profiles as well as $`c_s`$ and $`c_z`$ are now determined by the two variables $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$. The relation between $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$ and the experimentally controlled $`c_s,c_z,c_{\mathrm{DNA}}`$ is given by Eqs. 6-7. In terms of $`c_1^{\mathrm{}},c_z^{\mathrm{}}`$ the criterion for aggregation remains the same as in the previous case:

$$c_z^{\mathrm{}}=c_z^{}(c_1^{\mathrm{}}).$$
(11)
The three equations 6, 7 and 11, with the three unknowns $`c_1^{\mathrm{}}`$, $`c_z^{\mathrm{}}`$ and $`c_z`$ lead to a unique solution for $`c_{z,\mathrm{aggr}}`$. Note that $`c_1^{\mathrm{}}`$ is larger than $`c_s`$ because of counterions coming from the DNA as can be seen in Eq. 7, where $`a\rho _11`$ is negative. In Eq. 10, $`c_s`$ is replaced by $`c_1^{\mathrm{}}`$, which is larger than $`c_s`$ for large $`c_{\mathrm{DNA}}`$. Hence, increasing $`c_{\mathrm{DNA}}`$ has an effect similar to addition of monovalent salt. As noted above, this effect is significant only for $`c_{\mathrm{DNA}}>c_s`$.

## COMPARISON WITH EXPERIMENT

Raspaud et al. (1998) measured the spermine ($`z=4`$) concentration $`c_z`$ at the onset of aggregation as a function of $`c_{\mathrm{DNA}}`$ for four values of $`c_s`$ and with $`c_{\mathrm{DNA}}`$ ranging over four orders of magnitude — from $`10^2`$ to $`10^2\mathrm{mM}`$. We fitted the data (E. Raspaud and J.-L. Sikorav, private communication) for each $`c_s`$ to a straight line according to Eq. 10. The least square fit presented in Fig. 5 takes into account the experimental error bars and data points up to $`c_{\mathrm{DNA}}=10\mathrm{mM}`$. In Fig. 5 a the fit is shown using a linear scale which covers the range of $`c_{\mathrm{DNA}}`$ only up to $`c_{\mathrm{DNA}}=1.5`$ mM for clarity purposes. Due to the large range of $`c_{\mathrm{DNA}}`$ it is impossible to show all the data on the linear scale of Fig. 5 a. Instead, the same data and linear lines are shown in Fig. 5 b on a log-log scale over the full experimental range of $`c_{\mathrm{DNA}}`$.

The linear fit is very good for all four values of monovalent salt concentration $`c_s`$. Note that for $`c_s=88\mathrm{mM}`$ the fit is very good up to the largest value of $`c_{\mathrm{DNA}}=48`$ mM reported in the experiment, although our fit takes into account only data points up to $`c_{\mathrm{DNA}}=10`$ mM. It was previously suggested Raspaud et al. (1998) that a separate regime exists for $`c_{\mathrm{DNA}}10\mathrm{mM}`$, characterized by a power law relation between $`c_z`$ and $`c_{\mathrm{DNA}}`$ with an exponent smaller than unity. Our analysis suggests a different conclusion. The fit clearly demonstrates that the relation is linear all the way up to $`c_{\mathrm{DNA}}=48`$ mM, as predicted by Eq. 10. Note also that even at $`c_{\mathrm{DNA}}=48`$ mM we have $`c_{\mathrm{DNA}}<c_s`$ so the assumptions leading to Eq. 10 are still valid.

The only points in Fig. 5 b that deviate significantly from the fit are the three points where $`c_s=13\mathrm{mM}`$ (triangles) and $`c_{\mathrm{DNA}}>20\mathrm{mM}`$ (two of these points coincide with points having $`c_s=88\mathrm{mM}`$, shown using square symbols.) This deviation is easily explained by the fact that $`c_{\mathrm{DNA}}c_s`$ so that corrections to $`c_1^{\mathrm{}}`$ must be taken into account. For example, at $`c_{\mathrm{DNA}}=90\mathrm{mM}`$ the nominal monovalent counterion concentration is $`103\mathrm{mM}`$, taking into account counterions contributed by the DNA. In order to find $`c_1^{\mathrm{}}`$ we need to subtract the condensed counterions, as determined by $`\rho _1`$. We can estimate $`\rho _1`$ at this point by solving the Poisson-Boltzmann equation in a unit cell with the appropriate radius. The chemical potentials of the three ion species are tuned such that their concentrations match the known values of $`c_z`$ and $`c_s`$. This leads to an estimate: $`c_1^{\mathrm{}}68\mathrm{mM}`$. Hence, $`c_z`$ at the onset of aggregation should lie a little below the continuation of the $`c_s=88\mathrm{mM}`$ line which is, indeed, where it is found. The trend for $`c_s=13\mathrm{mM}`$ can probably be seen already at the point $`c_{\mathrm{DNA}}=15\mathrm{mM}`$, although the deviation at this point is still within the range of experimental error. The few other experimental points with $`c_{\mathrm{DNA}}c_s`$ deviate slightly from the straight line as well (still within experimental error bars). In all these cases the deviation is in the direction corresponding to a higher value of $`c_s`$, as expected.

A linear relation of the form $`c_{z,\mathrm{aggr}}=\alpha c_{\mathrm{DNA}}+\beta `$, was previously suggested on empirical basis for aggregation induced by spermidine ($`3^+`$), on a smaller range of DNA concentrations Osland and Kleppe (1977); Pelta et al. (1996b). Although this result looks similar to our prediction on the onset of aggregation, it is not directly related to our analysis because $`c_{z,\mathrm{aggr}}`$ was taken in those works to be the transition midpoint. This is the point where half of the maximal precipitation of DNA is reached. Our analysis does not apply at the transition midpoint since it requires all the DNA segments to be well separated from each other. Indeed, the coefficient $`\alpha `$, related to the transition midpoint, was found in Osland and Kleppe (1977) and Pelta et al. (1996b) to be of order $`10^2`$, much larger than unity. Such a value of $`\alpha `$ cannot be interpreted as the excess of spermidine ions per monomer near isolated chains.

The parameters of the linear fit in Fig. 5 are summarized in Table 1 for the four experimentally used values of $`c_s`$.

### Crossover in the log-log plot

For presentation purposes we plot in Fig 5 b, $`c_{z,\mathrm{aggr}}`$ vs. $`c_{\mathrm{DNA}}`$ on a log-log scale, as appeared in Raspaud et al. (1998). The linear relation that was found between these two quantities is not clearly manifested on the log-log plot, because a linear dependence of the form $`y=\alpha x+\beta `$ is not easily recognized in such a plot. Furthermore, such a linear relation appears on a log-log plot to be artificially characterized by two distinct behaviors, at low and high values of the independent variable. These two behaviors were mentioned in Raspaud et al. (1998) and can be seen in Fig. 5 b. However, they do not represent in our opinion two real physical regimes and can be understood by taking the logarithm of Eq. 10. For small $`c_{\mathrm{DNA}}`$ (large $`R`$):

$$\mathrm{log}c_z\mathrm{log}c_z^{}$$
(12)
i.e, $`c_z`$ does not depend on $`c_{\mathrm{DNA}}`$ as is seen in Fig. 5 b in the small $`c_{\mathrm{DNA}}`$ limit. In the opposite limit of large $`c_{\mathrm{DNA}}`$ (small $`R`$):

$$\mathrm{log}c_z\mathrm{log}c_{\mathrm{DNA}}+\mathrm{log}a\rho _z^{}$$
(13)
Here, the linear dependence of $`c_z`$ on $`c_{\mathrm{DNA}}`$ yields a line of slope 1 in the same figure.

The crossover between these apparent behaviors occurs when the number of bulk and excess ions are the same:

$$c_{\mathrm{DNA}}=\frac{c_z^{}}{a\rho _z^{}}$$
(14)
When $`c_{\mathrm{DNA}}`$ is much smaller than this crossover value, the number of excess multivalent ions near DNA segments is negligible compared to their total number. In the other extreme of $`c_{\mathrm{DNA}}`$ much larger than the crossover value, the number of free multivalent ions is negligible compared to the excess ions, and nearly all multivalent ions are bound to the DNA.

For the experimental data in Fig. 5 the crossover value is equal to $`0.06`$, $`0.18`$ and $`3.9\mathrm{mM}`$ for $`c_s=13`$, $`23`$ and $`88\mathrm{mM}`$, respectively, and smaller than $`1.5\times 10^3\mathrm{mM}`$ for $`c_s=2\mathrm{mM}`$. The first three crossover points are indicated by arrows in Fig. 5 b.

## DNA AGGREGATION AND COUNTERION CONDENSATION

We separate the discussion following our results in three parts. The first addresses the conditions required for DNA aggregation. The coefficients of the linear relation in Eq. 10, $`c_z^{}`$ and $`\rho _z^{}`$, have a definite physical meaning. Their values, as extracted from the experimental data provide insight on these conditions. The second part deals with condensation of counterions on DNA (to be distinguished from condensation of DNA chains). The general relation $`\rho _z=\rho _z(c_1^{\mathrm{}},c_z^{\mathrm{}})`$ that was introduced in Eqs. 3 \- 4 is a property of counterion condensation on isolated chains. By extracting the values of $`\rho _z`$, $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$ at the onset of DNA aggregation, we can learn about exact density profiles of spermine around DNA, and compare our findings with approximations such as Poisson-Boltzmann theory. Finally, we comment on our main assumption, which was used in the theoretical considerations section.

### Conditions at the onset of aggregation

Most of the proposed theoretical models for inter-chain attraction and aggregation (see, for example, Olvera de la Cruz et al. (1995); Raspaud et al. (1998); Borukhov et al. (2002, 2001); Wittmer et al. (1995); Nguyen et al. (2000); Ha and Liu (1997); Arenzon et al. (1999)) regard the charged chain as surrounded by a layer of condensed ions which is usually modeled as a one-dimensional gas. This layer mediates an inter-chain attraction, and the models predict the number of condensed ions required to initiate aggregation of the chains. In the current work we do not address this theoretical problem, but rather concentrate on what can be inferred from the experimental results using the analysis presented in the previous section. This analysis provides insight on the conditions prevailing at the onset of aggregation. In particular, the excess $`\rho _z^{}`$ characterizes the number of condensed multivalent counterions that are present near each chain at the onset. Although in general the notion of condensed counterions is somewhat ill-defined, as it depends on which ions are regarded as bound to the DNA, we show in the Appendix that in our case it does have a reasonably well defined meaning. Furthermore, the number of condensed multivalent ions per monomer is practically the same as $`a\rho _z^{}`$.

The excess of multivalent counterions per monomer, $`a\rho _z^{}`$, is shown in Fig. 6 as function of $`c_s`$. All values are taken from Table 1, as extracted from the experimental data. The dashed line is a linear fit. Two different axis scales are used on the left and right of the plot. The left axis shows the value of $`a\rho _z^{}`$. The right one shows the part of DNA charge that is compensated by condensed multivalent ions, $`za\rho _z^{}`$, where $`z=4`$ for spermine. From the plot we deduce the following two conclusions:

1. The number of condensed multivalent ions (per DNA monomer) $`a\rho _z^{}`$ at the onset decreases as the monovalent salt concentration increases, with variation between $`0.19`$ and $`0.14`$. A possible reason for this trend may be that the bare electrostatic repulsion between chains is decreased due to increased screening. Hence a smaller number of multivalent ions is required in order to overcome this repulsion. The change in $`\rho _z^{}`$ may also be related to the competition between monovalent and multivalent ions in the aggregated DNA state.
2. The data indicates that there is no over-charging of the DNA by spermine at the onset (see also Nguyen et al. (2000)) since $`za\rho _z^{}<1`$. At higher concentration of spermine, beyond the threshold, we do not rule out the possibility of DNA over-charging, as was suggested by Nguyen et al. (2000).

Although $`\rho _z^{}`$ decreases with increase of $`c_s`$, it is of the same order of magnitude for all the $`c_s`$ values in Table 1. In contrast, $`c_z^{}`$ varies in Table 1 over more than three orders of magnitude. As was previously suggested Olvera de la Cruz et al. (1995); Raspaud et al. (1998), this large variation in $`c_z^{}`$ is a result of competition between monovalent and multivalent counterions. We discuss the relation between $`\rho _z^{}`$ and $`c_z^{}`$ to some extent in the following subsection. A more detailed analysis of this relation, emphasizing the role of competition between the two counterion species, will be presented in a separate publication (see also, Belloni et al. (1984); Wilson and Bloomfield (1979); Wilson et al. (1980)).

### Counterion condensation

We now turn to analyze the condensation of monovalent and multivalent ions around DNA. Each line in Table 1 provides a measurement of the excess $`\rho _z`$ at certain values of $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$. The general relation $`\rho _z(c_1^{\mathrm{}},c_z^{\mathrm{}})`$ is a property of counterion density profiles around isolated DNA segments. Hence, the data in Table 1 can be used to test any particular theory used to calculate such ion distributions.

The most simple model to consider is the Poisson-Boltzmann (PB) theory (see Andelman (1994); Oosawa (1971); Le Bret and Zimm (1984); Guéron and Weisbuch (1980)). In Table 2 we compare the excess predicted by PB theory with the experimental result, by solving the PB equation such that $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$ match the experimental values of $`c_s`$ and $`c_z^{}`$ from Table 1. The excess is then calculated from the PB density profile, and compared with the experimental value of $`a\rho _z`$ (equal to $`a\rho _z^{}`$ of Table 1). The DNA is modeled as a uniformly charged cylinder of radius $`d=10\text{Å}`$.

Inspection of the results in Table 2 shows that there is a reasonable agreement with experiment (within the error bars) for the three smaller values of $`c_s`$ = 2, 13, 23 mM. However, for $`c_s=88\mathrm{mM}`$ there is a $`30\%`$ deviation. The two data points with $`c_{\mathrm{DNA}}>10\mathrm{mM}`$ that were not taken into account in the linear fit of Fig. 5 suggest that $`\rho _z`$ is closer to the lower bound of the experimental error range, whereas the PB value is larger than the upper bound.

Overall, the agreement with PB theory (Table 2) is surprisingly good considering that PB theory does not work so well for bulky multivalent ions. Deviations from PB theory have several sources. One of these sources is specific molecular details such as the geometrical shape of ions, DNA structure and short–range interactions. Another source for deviations are ion-ion correlations between spermine molecules, computed in theories which go beyond the mean-field approximation. However, these correlations tend to increase the number of bound multivalent counterions Lyubartsev and Nordenskiöld (1997), while for $`c_s=88\mathrm{mM}`$, the number of bound multivalent counterions is decreased. We conclude that correlation effects by themselves are not the main source of the deviations seen in Table 2. In addition the data analysis does not indicate over-charging of the DNA. Such an effect may be expected if correlation effects are strong Nguyen et al. (2000).

In Fig. 7 we compare the DNA aggregation data with PB predictions at finite DNA concentrations. For each DNA concentration the PB equation is solved in a cylindrical cell of the appropriate radius. The multivalent counterion concentration $`c_z`$ is gradually increased until the onset is reached, and its onset value, $`c_{z,\mathrm{aggr}}`$ is plotted as function of $`c_{\mathrm{DNA}}`$. Two different criteria are used to determine the onset $`c_{z,\mathrm{aggr}}`$. In Fig. 7 a it is chosen as the point where $`c_z^{\mathrm{}}`$ is equal to the experimental value $`c_z^{}`$ of Table 1; whereas in Fig. 7 b the onset is chosen the point where $`\rho _z=\rho _z^{}`$. In order to span all the data range we use for convenience a log-log plot, as in Fig. 5 b.

On a linear scale all the lines in Fig. 7 a and b are straight lines. This fact serves as additional confirmation of our general analysis in the theoretical considerations section. In accordance with our analysis, both $`c_z^{}`$ and $`\rho _z`$ are constant along each line, and the slope of each line is equal to $`a\rho _z`$. Note that the relation between $`c_z^{}`$ and $`\rho _z`$ is determined in Fig. 7 within the PB approximation, while in Fig. 5 both of these coefficients are related to the actual counterion density profiles in the experimental system. The use of the PB equation is the source of deviations from experimental data in Fig. 7.

On first inspection the match with experiment in Fig. 7 a is very good, whereas the match in Fig. 7 b is not as good. On closer inspection it is seen that the fit in Fig. 7 b is not good for small values of $`c_{\mathrm{DNA}}`$, while it is actually better than in Fig. 7 a for large $`c_{\mathrm{DNA}}`$. With the PB equation it is not possible to obtain a perfect fit for both small and large $`c_{\mathrm{DNA}}`$ because the values of $`c_z`$ and $`\rho _z`$ are not independent. Fixing $`c_z^{\mathrm{}}=c_z^{}`$ (as in Fig. 7 a) sets a value of $`\rho _z`$ that is different from $`\rho _z^{}`$; and the opposite happens in Fig. 7 b. The fit in Fig. 7 a is quite good even for large $`c_{\mathrm{DNA}}`$ because the values of $`\rho _z^{}`$ are of similar order of magnitude for all four lines.

Deviations as in Fig. 7 are inevitable if any approximations are used to model the distribution of counterions around DNA. Note however that within such approximate models our general theoretical considerations should apply, as long as the total number of ions in the system is counted properly. Such a model that goes beyond PB was proposed in Nguyen and Shklovskii (2001). Indeed, within this model a linear relationship similar to Eq. 10 was found.

The experimental results analyzed in this section may be influenced, to a certain degree, by the fact that there was more than one type of monovalent counterion in the system. For the three higher salt concentrations, except for $`c_s=2`$ mM, the solution contained $`10\mathrm{mM}`$ of TrisH<sup>+</sup> ions in addition to Na<sup>+</sup> Raspaud et al. (1998). For the largest salt concentration, $`88\mathrm{mM}`$, where significant deviations from PB theory are found, this effect is probably negligible. Another detail regarding the TE buffer is that the Tris ions may be only partly ionized. If only 80% of Tris is ionized, as suggested in Tang et al. (1997), the concentrations $`c_s=13`$ mM, 23 mM and 88 mM should be reduced by 2 mM. Although this will have only a small effect on our results, it will improve both the comparison with PB and the fit with the dashed line in Fig. 6, for the point $`c_s=13`$ mM. For the two other concentrations of 23 mM and 88 mM the effect will be negligible.

### Further comments on underlying model assumption

Our underlying assumption, that the onset of aggregation depends uniquely on $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$ (but not on $`c_{\mathrm{DNA}}`$), is an approximation that can be justified on several different levels but deserves further and more thorough investigation. The most simple motivation for this assumption is that multivalent ions, in the vicinity of the chains, mediate the attraction necessary for aggregation. In turn, the number of condensed multivalent ions near each chain is determined by $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$.

Let us first suppose that aggregation starts when a net attraction appears between two chains. This assumption may be justified if chains are sufficiently long and their translational entropy can be neglected. In order to find the onset of two-chain attraction the free energy of a two-chain complex should be calculated as a function of the distance between the two chains. This free energy represents the effective interaction between the two chains, mediated by the ionic solution. The counterion distribution near each chain will not be the same for close-by and for isolated chains. However in both cases the concentrations must decay to their bulk values throughout the solution, $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$. This requirement serves as a boundary condition, imposed at a large distance from the two chains. It will determine uniquely the counterion distribution between the chains, as well as the free energy associated with the two-chain complex. Hence $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$ determine the effective interaction between chains, and in particular whether an attraction occurs at a certain range of inter-chain separations; in terms of these variables the onset of two-chain attraction does not depend on $`c_{\mathrm{DNA}}`$.

Strictly speaking, the onset of aggregation and the onset of two-chain attraction are not the same. The aggregate phase involves interactions between multiple chains, whereas chains in the dilute phase interact very weakly with each other. Aggregation starts when the free energy per chain is equal in the dilute and aggregate phases. Note that the chemical potential of each ion species must be the same in the two phases, and that in the dilute phase these chemical potentials are directly related to $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$. Hence $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$ determine the free energy per chain in the two phases. The approximation of independence on $`c_{\mathrm{DNA}}`$ neglects the translational entropy of DNA segments, which can be justified for long enough and rigid segments. It also neglects contributions from interactions between chains in the dilute phase, which are assumed to be small compared to the free energy of the single DNA-counterion complexes.

## SUMMARY

We have shown that the onset of aggregation at finite (non-zero) DNA concentration, $`c_{z,\mathrm{aggr}}`$, is determined by the onset in the limit of infinite DNA dilution. For DNA monomer concentration smaller than that of monovalent salt, $`c_{\mathrm{DNA}}c_s`$, the multivalent counterion concentration at the onset, $`c_{z,\mathrm{aggr}}`$, depends linearly on $`c_{\mathrm{DNA}}`$. The coefficients of this linear dependence are the bulk concentration of multivalent counterions and their excess relative to the bulk near each DNA segment. Both of these coefficients are of theoretical interest and can be extracted from the available experimental data.

Our main assumption is that the onset of aggregation can be related to the ion density profiles around each chain. Hence, it is uniquely determined by $`c_1^{\mathrm{}}`$ and $`c_z^{\mathrm{}}`$, the bulk concentrations of the two counterion species, respectively. Our results and fit to experiment strongly support this assumption. Nevertheless, we believe that more detailed theoretical and experimental investigations are needed in order to fully understand its range of validity. For example, it will be of interest to test experimentally the equilibration of a DNA solution through a dialysis membrane, with a cell containing only counterions Braunlin et al. (1982); Plum and Bloomfield (1988); Subirana and Vives (1981). This procedure allows a direct control of the ionic bulk concentrations.

In order to predict precisely the onset of aggregation, the structure of the aggregated phase must be considered. Nevertheless, it is instructive to focus only on single chains at the onset, as is often done. At the aggregation onset the electrostatic repulsion between isolated chains in solution must be overcome by a sufficiently strong attraction mediated by multivalent counterions. This number of counterions is expected to depend only weakly on physical parameters such as the monovalent salt concentration. Our analysis does not address directly the question of the onset origin, but merely supports the fact that the number of condensed multivalent ions at the onset, $`a\rho _z^{}`$, is of the same order of magnitude, regardless of the $`c_s`$ value. A more refined result of our analysis is that $`a\rho _z^{}`$ is not constant but decreases with increase of $`c_s`$. On the other hand $`c_z^{}`$, the value of $`c_z^{\mathrm{}}`$ at the onset, depends strongly on $`c_s`$. This is mainly a result of the competition between monovalent and multivalent ions, as will be addressed in a separate publication.

Our analysis also sheds light on counterion condensation on DNA, which is independent on the criterion for DNA aggregation. The experimental data indicates that for high $`c_s`$ the number of spermine ions in the vicinity of DNA is smaller than the prediction of Poisson-Boltzmann theory. A similar trend was observed in computer simulations Lyubartsev and Nordenskiöld (1997) of spermidine (3<sup>+</sup>) and NaCl in contact with DNA. Spermidine binding was affected by addition of monovalent salt more strongly than the Poisson-Boltzmann prediction. For high salt concentrations spermidine binding was considerably smaller. In the computer simulations both molecular specific interactions, the geometrical shape of the constituents and inter-ion correlations were taken into account. All these effects, and in particular the geometry of the spermidine molecule, which is similar to that of spermine, were found to play an important role.

The above analysis demonstrates that specific interactions play an important role in determining the threshold of aggregation. In the dilute phase these interactions strongly influence the competition between monovalent and multivalent ions and the free energy of DNA-counterion complexes. Similarly, specific interactions play a prominent role in the dense phase Strey et al. (1998). Force measurements under osmotic stress Rau et al. (1984); Rau and Parsegian (1992a, b) provide a wealth of information on these interactions.

In conclusion, the physical parameters extracted here from experiment on the onset of DNA aggregation provide insight on the conditions required for aggregation, and on condensation of ions around DNA. These parameters may turn out to be of great value in assessment of various theoretical models. Additional detailed experiments may further deepen our understanding of these complex phenomena.

We are grateful to E. Raspaud and J.-L. Sikorav for numerous discussions and for providing us with their unpublished experimental data. We also wish to thank I. Borukhov, H. Diamant, M. Kozlov, A. Lyubartsev, G. Manning, T. Nguyen, M. Olvera de la Cruz, A. Parsegian, R. Podgornik, D. Rau and T. Thomas for discussions and correspondence. Support from the U.S.-Israel Binational Science Foundation (B.S.F.) under grant No. 98-00429 and the Israel Science Foundation under grant No. 210/02 is gratefully acknowledged. One of us (DA) thanks the Alexander von Humboldt Foundation for a research award.

## APPENDIX

In this appendix we discuss the relation between the excess and the number of condensed ions. The latter quantity is not as well defined as the former, but relates more naturally to the aggregation mechanism. The notion of condensed ions suggests that some ions are bound to the charged chain while others are free. In reality there is a density profile that extends all the way from $`r=d`$ to $`r=R`$ with no definite separation between condensed and free ions. In the following we define condensed ions rather loosely as the number of ions up to a certain characteristic distance from the chain Belloni et al. (1984); Wilson et al. (1980). We show that for multivalent ions this number does not depend strongly on the choice of this characteristic distance. Hence, the number of condensed ions is reasonably well defined. Moreover, the excess number of multivalent counterions, which can be directly calculated from the experimental data, is nearly identical to this quantity. This point will be further explained below.

Fig. 8 shows the excess of 4-valent counterions $`\delta \rho _z(r)`$ up to a distance $`r`$ from the DNA axis, as a function of $`r`$:

$$\delta \rho _z(r)=2\pi _0^rr^{}dr^{}\left[n_z(r^{})c_z^{\mathrm{}}\right]$$
(A1)
with the limit $`\delta \rho _z(\mathrm{})=\rho _z`$ of Eq. 4. The density profile was calculated using the Poisson-Boltzmann equation, with the radius of DNA taken as $`d=10\text{Å}`$ and with bulk densities of ions as in the last line of Table 1: $`c_s=c_1^{\mathrm{}}=88\mathrm{mM}`$, $`c_z^{\mathrm{}}=0.52\mathrm{mM}`$.

Three observations can be made. First, most but not all of the excess z-valent ions are localized very close to the DNA, at a distance of order $`\lambda /z`$, where $`\lambda `$ is the Gouy-Chapman length (see Andelman (1994)):

$$\lambda =\frac{1}{2\pi l_B\sigma }=\frac{d}{l_B\rho _{\mathrm{DNA}}}$$
(A2)
where $`\sigma `$ is the average charge per unit area on the cylinder surface, $`\sigma =\rho _{\mathrm{DNA}}/2\pi d`$, and $`\rho _{\mathrm{DNA}}=1/a`$ is the DNA charge per unit length. At room temperature the Bjerrum length $`l_B7\text{Å}`$ and for DNA with 4-valent counterions $`\lambda /z0.6\text{Å}`$. Second, the counterions within a layer of few times the Debye length ($`\kappa ^1=10.0\text{Å}`$ in Fig. 8) neutralize the DNA charge. Nearly all the excess distribution is in this layer. Third, in order to estimate the total amount of counterions in the condensed layer of thickness $`\alpha \kappa ^1`$, where $`\alpha `$ is a number of order unity, we need to add $`\delta \rho _z`$ to the bulk contribution, $`\pi \alpha ^2\kappa ^2c_z^{\mathrm{}}`$. Using $`\kappa `$ from Eq. 1, the latter is equal to:

$$\left(\frac{\alpha ^2}{4l_B}\right)\frac{c_z^{\mathrm{}}}{2c_1^{\mathrm{}}+z(z+1)c_z^{\mathrm{}}}$$
(A3)
In experiment, $`c_z^{\mathrm{}}`$ is much smaller than $`c_1^{\mathrm{}}`$ at the onset, and the bulk contribution of Eq. A3 can be neglected relative to $`\rho _z`$, for $`\alpha `$ of order unity. This can be seen specifically in Fig. 8 by comparing the solid and dashed lines.

The outcome of the above discussion is that $`\rho _z`$, defined in Eq. 4 as the excess of counterions throughout the cell, can be regarded, to a good approximation, as the total number of counterions within a condensation layer whose thickness is approximately the Debye length. For typical concentration ranges as considered here we do not expect that this outcome will change, even for models going beyond Poisson-Boltzmann theory.

As a further demonstration, the number of multivalent counterions up to several different distances from the DNA is shown in Table 3, as calculated in a unit cell using the Poisson-Boltzmann equation. For each $`c_s`$ in Table 1 we find the Poisson-Boltzmann density profile such that $`c_1^{\mathrm{}}=c_s`$ and $`\rho _z=\rho _z^{}`$, and then calculate the number of multivalent ions (per DNA monomer) up to the following distances from the DNA radius: 10 Å, 20 Å, $`\kappa ^1`$ and $`2\kappa ^1`$. The values of $`\kappa ^1`$, as obtained from Eq. 1 are equal to 68, 26, 20 and 10 Å for $`c_s=`$ 2, 13, 23 and 88 mM, respectively. These numbers are compared with $`a\rho _z^{}`$. All the different measures in Table 3 yield results that are very close to each other.