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To cite this article: Justin E. Molloy & Miles J. Padgett (2002) Lights, action: Optical tweezers, Contemporary
Physics, 43:4, 241-258, DOI: 10.1080/00107510110116051
To link to this article: http://dx.doi.org/10.1080/00107510110116051
1.
Introduction
2.
2.1.
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Contemporar y Physics ISSN 0010-7514 print/ISSN 1366-581 2 online # 2002 Taylor & Francis Ltd
http://www.tandf.co.uk/journals
DOI: 10.1080 /0010751011011605 1
242
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2.2.
Figure 1.
The ray optical origin of the lateral and axial trapping force within optical tweezers.
2.3.
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2.4.
243
2.5.
q2 x
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244
1
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10
3.
3.1.
3.2.
245
4.
Multiple tweezers
4.1.
Figure 3.
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b
A simple optical relay system to give lateral control of the beam position in the sample plane of optical tweezers.
11
246
d 2Dt
1=2
12
4.2.
Figure 4. Video images of spheres held by four optical tweezers, which have been synthesized by rapidly scanning a single laser beam
using an acousto-optic beam deector. The single beam was time-shared between the diVerent locations at 10 kHz. Successive frames
show how the four trapped plastic microspheres 1 mm in diameter can be independently manipulated.
5.
247
248
Figure 5. Layout of a multiplexed dual optical tweezers transducer for making measurements of force and movement produced by
single molecules. The apparatus is based around a conventional research grade inverted microscope (e.g. Zeiss Axiovert). The optical
tweezers are produced by a diode-pumped neodymium-doped yttrium aluminium garnet (Nd:YAG) laser introduced using a light path
similar to gures 2 and 3. A dual dichroic mirror (DDM) is used to reect infrared and green light but to transmit red light from
uorochromes. Mechanical shutters (MS), dichroic mirrors (DM), excitation lters (EF) and barrier lters (BF) together with an
intensied charge-coupled device (ICCD) camera allow observation of uorescent proteins. Central to this particular design is a
computer that controls the piezoelectric transducer (PZT) on the microscope, the beam steering optics (acousto-optic deectors AOD)
and also collects data from the two four-quadrant imaging detectors (4QD) that monitor the position of both objects. Computer software
allows a closed feedback loop to control the bead positions to compensate for any detected motions.
determined using a four-quadran t photosensor to measure the `centre of gravity of the object, either using
conventiona l imaging (bright eld, dark eld or phase
contrast) or by interferometry. If a very intense
illumination source is used, then it is possible to measure
nanometre displacements over a bandwidth of 0.1 Hz to
5 kHz. The sensor and detection circuitry is calibrated by
moving either the sensor itself or the trapped objects
through a known distance. Once the position calibration
factor is known, then the optical tweezers stiVness can be
calculated either from analysis of the thermal motion of
the trapped object (knowing equation (9) [gure 6 (c)]) or
by applicatio n of known viscous drag forces (either from
spectral analysis, equations (8) and (10) [gure 6(b)] or
by applyin g a viscous drag force F bv). Usually, a
combination of approaches is used to check that they are
249
Figure 6. Operation of an optical tweezers transducer. (a) The motion of a plastic microsphere 1.5 mm in diameter held in optical
tweezers of k 0.018 pN nm 1. Note that the bead position uctuates on a relatively slow time scale (tens of milliseconds) because of
the high viscous damping and low tweezers stiVness. (b) Spectral analysis of the movement shows the expected Lorenzian behaviour with
characteristic single-pole roll-oV. (c) The histogram shows how the number of observations of the bead at position x is determined by a
Boltzmann distribution given by N expkx2 =2kB T.
250
251
Figure 7. Mechanical recording made from a single processive kinesin, a molecular porter that walks along the microtubule track
taking steps that are commensurate with the microtubule 8 nm lattice repeat. In the experiment a single (double-headed) kinesin
molecule was attached to a latex microsphere held in optical tweezers. (a) The position of the microsphere was monitored using a fourquadrant detector, as the kinesin walked along a xed microtubule track. (b) Note that the staircase structure to the position data is a
direct result of the single kinesin pausing in between individual ATP cycles. This record was kindly provided by Dr N.J. Carter and Dr
R.A. Cross, Molecular Motors Group, Marie Curie Research Institute, Oxted, Surrey. For further information, see http://
mc11.mcri.ac.uk/motorhome.html.
252
6.
253
Figure 8. Mechanical recording made from a single non-processive myosin, a molecular rower that interacts intermittently with actin.
(a) The actin lament and myosin must be positioned close to one another so that multiple individual events can be observed. Myosin is
coated on a large glass microsphere xed to a microscope coverslip, while actin is suspended between two beads held using a dual optical.
By monitoring the position of one or both beads held in the optical tweezers, individual binding events can be observed. (b) Note that the
thermal noise is reduced during binding events because the actomyosin bond forms a stiV link to mechanical ground.
Figure 9. The various schemes employed to achieve rotation of a particle within optical tweezers; transfer of (a) spin and (b) orbital
angular momentum of the light beam to the trapped particle, torque generated by the gradient force created by (c) rotation of a highorder mode, (d) rotation of an asymmetric interference pattern or (e) rotation of an aperture and nally (f) the radiation pressure acting
on propeller-shaped objects (see text for further details).
254
Figure 10. The gradient force associated with a rectangularly symmetric mode will create a torque on an asymmetric cell, causing
alignment of the cell with the beam.
255
6.2.
256
which have a lower refractive index than their surrounding media, the direction of the gradient force is reversed.
Consequently, rather than being attracted to the highest
intensity region, the low-index particles are repelled from
the beam axis. Use of a Laguerre Gaussian mode
overcomes this problem because, once within the annular
ring, a low-index particle experiences a gradient force
again directed to the beam axis. Called `vortex traps,
optical tweezers based on Laguerre Gaussian modes have
enabled the three-dimensional connement of hollow glass
spheres between 2 and 50 mm in diameter [52, 53]. It is
interesting to note that, for a beam directed downwards,
the stable trapping position for low index spheres lies just
above the beam focus. The same group demonstrated that
these vortex traps could simultaneously conne high-index
particles to the normal position of just below the beam
focus.
In his early considerations of the trapping mechanism,
Ashkin pointed out that, since the trapping force results
from a change in ray direction, it is only the oV-axis rays
that contribute to the axial trapping force. Indeed, his
calculation s predicted that modest improvements in Qaxial
were possible using a TEM01 `doughnut mode, which has
no on-axis rays. With their zero on-axis intensity,
Laguerre Gaussian modes also satisfy this condition.
Indeed recent experimental results conrm that high-order
Laguerre-Gaussian modes do, at least for spheres above
2 mm in diameter, improve the axial trapping e ciency of
optical tweezers [54].
A further recent development in optical tweezers is the
use of Bessel light beams. Bessel beams [55], which are
frequently referred to as `diVraction free beams, consists of
a bright central spot surrounded by concentric rings of
Figure 13. Schemes employed for trapping metal particles: (a) submicron particles trapped at the beam focus by the force arising from
light scattering; (b) Mie particles trapped by the surface plasmon wave; (c) larger particles trapped by rotating or (d) annular beams to
conne that particle or (e) by light scattering around the outside of a Laguerre Gaussian mode.
central core of the beam (gure 12). In this case the Bessel
beam was created from a standard Gaussian beam using a
specially fabricated glass axicon, although computergenerated hologram s could also be used for the same
purpose.
6.4.
6.5.
7.
257
Acknowledgements
M.J.P. and J.E.M. both thank the Royal Society for
establishing their research careers by the awards of
University Research Fellowships.
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