Study of rotational dynamics of

trapped asymmetric particles in optical

tweezers
SpontaneousRevolutionofMicro
swimmersinsideAnnularTrap
We used an inverted microscope, where a Laser (Lasever Technologies, wavelength 1064 nm
with a linewidth of less than 1 nm, maximum power output 600 mW) that is checked to have
a TEM00 (Gaussian) output mode is coupled to the microscopes through the back port. The
basic setup schematic is shown in Fig.4.1. In general a Laser power of around 500 mW is
used for trapping. Because of power loss from the coupling optics the typical power that goes
past the microscope objective inside the sample chamber is around 40% of the laser power
that is coming out of the source. The optical tweezers is setup around a Carl Zeiss Axiovert
Observer-A1 microscope. A CCD camera attached at its side port is used for imaging. A
motorized sample stage is used in order to scan the sample when required. The laser beam
passes through a beam expander that resizes the beam so that it overfills the back aperture of
the objective lens, followed by a beam steering arrangement that allowed to center and make
the beam perpendicular to the plane of the sample. We use Carl Zeiss 100x, PlanApochromat, Infinity corrected objective Lens that has NA of 1.41. The trapping laser is
focused inside the sample chamber by varying the objectives distance from the sample. The
sample consists of a dilute solution of a peapod-shaped soft oxometalate particle, which is
loaded with a catalyst, in distilled water. The shape of the particle is not symmetric. The
solution is placed between a glass slide and a cover slip. The typical working distance of this
objective is 170m, however, we observed that we could focus into our sample chamber at
distances around 300m from the exit pupil of the objective. For this particular experiment
our set-up varies from the conventional setup in the sample loading arrangement in the
following ways:
1. The cover-slip thickness and material.2. The spacing between the glass slide and the coverslip.3. The amount of sample loaded inside the sample chamber.
In this arrangement the sample was compressed between a glass slide having a refractive
index of 1.516, and a cover-slip. This was then placed directly above the microscope
objective in contact with the immersion oil. This is shown in Fig.4.1. The cover-slip that we
use is made out of a polymer (Sigma Aldrich Hybrid Cover-slips, Part no. Z365912-100EA)
having a refractive index of 1.572 (at 1064nm) and had a thickness of 250m. The refractive
index of the immersion oil was also around 1.516, hence there is a significant refractive index
mismatch with the immersion oil. For certain focusing. In the experiment we trapped a
peapod-shaped soft oxometalate particle, which is loaded with a catalyst, inside the ringshaped optical trap. The particle was observed to rotate along the periphery of the ring shaped
potential. One such rotation event can be seen in the image sequence in Fig.4.2. The mode of
rotation was not specific and the occurrence of clockwise or counterclockwise rotation was
random. The radius of the annular trap can be varied from 3.2m to 5m the particle was seen
to follow the modified ring potential. However the rotational frequency decreases with the

increase in the ring diameter.

The input polarization of the Laser was linear and has no intrinsic angular momen- tum in it.
Therefore there is no possibility of angular momentum transfer CXXX for which the peapod
happened to rotatate inside the annular trap. One way to explain such revolution is because of
imbalance in the scattering of the input electric field from the surface of the particle it gets a
non-zero resultant force component along one direc- tion. If this force is tangential to the
perimeter of the ring potential, the particle will rotate inside it and it will never come out of
the trap. Another important to remember is that the force has to overcome the viscous drag
force in order to start revolving. The lowering in rotational frequency can be attributed to the
redistribution of the constant input power into a bigger ring. The change in intensity leads to a
different amount of scatter from the peapod. The rate of revolution can be controlled from a
few Hz to tens of Hz by changing the intensity of the trapping light which can be achieved
either by modifying the laser power or the annular trap diameter. In order to confirm whether
the scattering force is the governing body for such revolution of peapods inside the annular
trap we have performed a theoretical simulation of our physical system. In summary the
problem is to propagate a highly focused Gaussian beam through a stratified medium, which
comprises of three layers, i.e., (1) Oil with thickness around 5m, (2) Coverslip of 250m
thick, and on top of it is sample chamber of 25m in height. The peapod shaped particle has
been mimicked with a truncated cone like structure as shown in Fig. FXXX. Our goal is to
calculate the scattering force acting on the particle and compare it with the drag force.
Numerous methods have been proposed to solve Maxwells equations in media under many
approximations. However, all methods can be classified broadly into two regimes - the time
dependent and the time independent. In certain class of problems like light pulse propagation
in optical fibers, time domain methods are used, whereas in other problems like determining
standing wave modes of an optical cavity, time independent methods are more useful.
Classifications can also be done based on the form of approx- imations that are used to solve
the equations like paraxial or small angle approximation methods, and also the type of
mathematical techniques that are used to solve such problems - Fourier methods, propagator
methods, grid-based or finite element methods.

Shape Anisotropy Induced Spinning of

Red Blood Cells Inside an Optical Trap
The experimental setup was same as mentioned in Sec. 4.1 except the sample chamber
configuration. Unlike previous experiment we used 160m glass coverslip below the glass
slide. The refractive index of the coverslip was same as the immersion oil used. Therefore no
ring potential was seen to be formed. Normally the size of an RBC varies around 4 5m and
as they are heavy enough to get precipitated with in few seconds and attach with the lower
surface. In order to circumvent this problem we prepared a bigger sample chamber by putting
double sided tape in between the cover slip and the glass slide. We can load up to 50l of
sample inside such chamber. It helped us to continue the experiment for longer duration. The
sample was prepared in normal salt water. We collected about 50l blood sample from a
healthy donor (it was the author) and diluted it in 1ml of saline (0.9%w/vNaCl + H 2O) water
to prepare the stock solution. 10l of anti-coagulating agent Heparin was added into this stock
solution to prevent clumping of RBC. The solution was further diluted by mixing 200l of the

stock solution into 1ml of 2%w/v salt water solution. The sample was kept at 4 C so that RBC
does not disintegrate with in a day. 1 hour after preparing the diluted solution in hypertonic
solvent, we start the experiment. By that time the RBCs get deformed as the water flows out

of the cell to maintain the concentration equilibrium. Because of shape anisotropy we

observed RBCs to spin inside the optical trap. The rotational speed increases with the increase
in laser power. The deformed RBCs were found to rotate inside the optical trap, where as no
rotational dynamics was observed in case of healthy normal RBCs. Two possible explanations
were given by Mathur et.al. CXXX. One is because of shape anisotropy of the deformed
RBCs it gives rise to asymmetric scattering, which led to generation of non-zero torque.
Another hypothesis was due to change in shape the birefringence of RBCs CXXX in- crease
rapidly. As the input laser source is linearly polarized, any misalignment between input
polarization and the optical axis of the RBC produces a torque. It tries to reorient the RBCs
optical axis along the polarization axis of the input beam. Such an initial kick gives rise to
rotational dynamics, which is governed by many other factors like hydrodynamic interaction
CXXX. But as we have already pointed out, while explaining the dynamics of microswimmers, in optical tweezers we are dealing with objects that are in length scale of m. At
this stage the system can be assumed to be over damped. Therefore kick or to be specific
inertial force cannot be held as possible explanation for any dynamics inside the optical trap.
We conclude that the imbalanced scattering is the major reason behind the rotation of RBC
when it is trapped by optical tweezers. In order to confirm our conjecture we simultaneously
executed the numerical simulation using Lumerical and performed the experiment. We
simulated the coupled Brownian motion in Matlab using Eqn. 5.1, and 5.2 that gave us
computer generated Brownian motion of a particle inside a trap. The cross- correlation of
translational Brownian with rotational Brownian motion was studied for different coupling
constant. From Fig. FXXX it can be seen that as the coupling increases the cross correlation
amplitude tends to be a rising linear function. Also Fig. FXX shows two auto-correlation
function between rotational and translational Brownian motion for two different coupling
1
constant of = 6.9 rad/s Fig. FXXX(A), and = 1.9rad/s Fig. FXXX(B). The dotted line
in Fig. FXXX shows the best fitted curve using Eq. 5.4. The best way to acquire Brownian
motion data in experiment is to use Quadrant Photo Diode (QPD). The basic working
principle can be found in many literature CXXX. But using the traditional method one can
only get the translational Brownian motion from a QPD. We used a QPD taken from a CD
player head (chip Sony KSS-213C) (see Ref. CXXX for details). It gives independent voltage
outputs for each of the 4 quadrants. The newly developed method in our lab CXXX to detect
rotational Brownian motion was used in this experiment for data acquisition. Both the
rotational and translational data acquisition was done at a sampling frequency of 40KHz taken
for 5 seconds using PCIe-6361 data card. The cross-correlation between the translational and
rotational motion of a deformed RBC as shown in Fig. FXXX, matches with our numerical
simulation. Therefore it confirms that the Brownian motion of a deformed RBC inside an
optical trap is a combination of rotational and translational motion as they are not uncoupled.
The rate of decay of correlations depend on two factors: (A) the strength of the white noise in
the system, and (B) the regularity of the observables. Therefore the amplitude and the decay
rate gives an important information about the shape anisotropy of the RBC because of its
change in rheology due to the increase in osmolarity. To nullify the idea the we are getting
spurious results because of experimental artifacts we checked the cross correlation of
rotational and translationl Brownian motion of a 3m polystyrene bead trapped using the
same optical tweezers. As a polystyrene bead is spherical and there is not presence of shape
anisotropy that can couple the Brownian motions, they should be uncorrelated and will give a
constant value equal to zero. The amplitude of the cross-correlation data for a bead was 100
times less than RBC data. As a remark we can state that in hypertonic salt solution RBCs get
deformed and gives rise to shape anisotropy. The confirmation for that is the coupled rotationtranslation Brownian motion inside an optical trap. But this does not confirm whether shape
anisotropy plays the mojor role behind spinning of RBC inside optical tweezers. In the
following section we showed that shape anisotropy induced form birefringence is not enough
to generate a net torque in order to spin the RBC.