Optical tweezers

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Optical trapping
Photons carry momentum; consequently when light
shines on a surface there is an effective force exerted
on that surface.
Usually this effect is very small and can be neglected.
For intense light shining on a small particle, forces in
the range of 1 to 200 pN can be generated.
An extension of this concept is to create a specially
focussed light beam that creates a potential well that
traps a bead or small particle, typically 12 m in
diameter
 Physics of optical tweezers
Dielectric objects are attracted to the center of the
beam depending on the refractive index mismatch to
physically hold and move microscopic dielectric
objects
The force applied on the object depends linearly on
its displacement from the trap center just as with a
simple spring system.
A generic optical tweezer diagram
 Optical tweezers
Optical tweezers are capable of manipulating nanometer
and micron-sized dielectric particles by exerting extremely
small forces via a highly focused laser beam.

The beam is typically focused by sending it through a

microscope objective.

The narrowest point of the focused beam, known as the

beam waist, contains a very strong electric field gradient.

It turns out that dielectric particles are attracted along the

gradient to the region of strongest electric field, which is
the center of the beam.
 Optical tweezers

Photons that are absorbed or scattered by the tiny

dielectric particle in its path impart momentum to
the dielectric particle.
p = mv

This is known as the scattering force and results in

the particle being displaced slightly downstream
from the exact position of the beam waist
Schematic description of the optical tweezers manipulating a
bead that is attached to a cultured cell. Bead position is
monitored by microscopy and so the relative position of the
bead and the optical trap center are known.
 Optical tweezers
This principle can be used for biological measurements by
coating the bead with fibronectin (or some other
molecule that will bind to receptors on the cell surface)

The beam is then moved laterally, and the motion of the

bead is observed microscopically

From knowledge about the characteristics of the light

trap, the force exerted on the bead by the moving light
beam can be determined from the bead position relative
to the center of the optical trap.
 Optical tweezers
Since the displacement of the bead is simultaneously
monitored, this information can be used to determine the
local stiffness of the cell.

Essentially the same technique can be used to measure

the force exerted by individual molecules.

Optical trap techniques have been used for a variety of

biomechanical measurements
 The Optical Cell
Rotator is a fiber
based laser trap
that can hold and
precisely orient
living cells for
tomographic
microscopy.
 Magnetic bead microrheometry
In the magnetic bead microrheometry technique, a
paramagnetic bead, typically 45 m in diameter, is
coated with fibronectin or some other suitable
molecule.

The fibronectin binds to integrins on the cell surface,

providing a direct link between the bead and the
actin cytoskeleton of the cell

The paramagnetic bead is then subjected to a

magnetic field that either twists it (magnetic twisting
cytometry or displaces it (magnetic bead
microrheometry
 Schematic description of the central measuring unit of a magnetic
bead microrheometer based on an electromagnet.
The electromagnet is attached to a microscope stage and consists of
a coil (1200 turns of 0.7mm copper wire) and a soft iron core.
This core extends beyond the windings, forming a pole piece that
penetrates the sample chamber.
The tip of the pole piece can be positioned at distances of (r) of 10 to
100 m from a magnetic bead attached to a cell.
This produces maximal forces of 10 000 pN on a paramagnetic bead
of 4.5 m diameter.
 concentrate on magnetic bead
microrheometry, where the magnetically
induced motion of the bead is visualized by
light microscopy

Information about bead displacement and

applied force is used to estimate local
mechanical properties of the cell

One advantage of this approach is that it can

produce forces in the 10010,000 pN range,
which is relatively large.
 Calibration
An important issue is how the system will be
calibrated.
This is accomplished by suspending a bead in a liquid
of known viscosity and applying a magnetic field to
the bead.
The resulting displacement of the bead can be
tracked optically as a function of time, and then
Stokes law can be used to estimate the force on the
bead.
From this, the force profile as a function of applied
current and distance from the magnets pole piece
can be determined
 RESULTS
when a force is applied to the bead, there is a
nearly immediate displacement of the bead
(phase I ), followed by a gradual creep (phases
II and III).
This is consistent with the cytoskeleton and
cytoplasm demonstrating viscoelastic
behavior.
technique can be used to image the strain field
around a single paramagnetic bead
bound to a cell
displacement field induced by a paramagnetic bead is mapped on the
plasma membrane
of a fibroblast. (A) Microphotograph showing one magnetic bead (M,
radius 2.25 m) and several non-magnetic
particles (19, radius 0.5 m) attached to a region of the cell membrane
 This approach involves seeding the surface
of the cell with latex microspheres and
watching their displacement in the
neighbourhood of the magnetic bead.

This gives information about the length scale

over which displacements are coupled within
the cell.
 Structural cytoskeletal components within the cell
altering the local deformation field

By variations in the strength of attachment of the

small beads to the integrin receptors

By differences in the strength of the coupling

between the integrin receptors and the cytoskeleton