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Optical Tweezers
A revolution in
micro-manipulation
Jonathan [email protected]
k
University of Glasgow
Today’s talk
• What are optical tweezers?
• Dynamic movement and multiple particles
• Current research around the World
What are optical tweezers?
Optical tweezers use light to trap, manipulateand position micron sized objects.
Invented approximately 20 years ago by A. Ashkin et al.
K.C. Neuman and S. M. Block, Optical Trapping, Rev. Sci. Inst., (2004)
J. E. Molloy and M. J. Padgett, Lights, Action: Optical Tweezers, Cont. Phys., (2002)
What are optical tweezers?
A tightly focused laserproduces a force great enough to trap micron sized dielectric particles.
Require……1. Laser2. Lens3. Object4. Damping medium
Fscatt
Fgrad
Optical tweezers in action
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The equipment
Optical tweezers are based on high magnification microscope lenses
–produces tightly focussed beam–provides visualisation of image
Samples suspended in fluid
–provides cooling–provides buoyancy
The equipment
Require tight focusing so needhigh numerical aperture, N.A.
Magnification typically X100
N.A. = n sin()
n is the refractive index of the medium between the objective lens and the sample. Using oilimmersion lenses, n ~ 1.3 so N.A >1is possible.
Optical Trapping - a>> Conditions for Mie scattering when the particleradius a is larger than the wavelength of the light .
We can use a ray optics argument andlook at the transfer of momentum
a
Optical Trapping - a<< Condition for Rayleigh scattering when the particleradius a is smaller than the wavelength of the light .
Scattering force and gradient force are separable
Fgrad > Fscatt requires tight focusing
a
The scalesCan trap 0.1 to 10’s m
1m is…..…the same as 1/100th diameter of a hair.
In water, you can move a particle at about 20-30m per sec.
Require 10mW per trap.
Can rotate at 100’s of Hz.
If absorbed by particle of refractive index n, a beam of power P produces a reaction force
F = nP/c
(e.g. P = 1mW: F = 5pN)
The Q factor of optical tweezers
The efficiency Q, of optical tweezers is defined as
Q = Factual/ (nP/c)(typically Q ≈ 0.05-0.3)
Optical Trap Dynamics
Equation of motion of particle in a potential well
restoring force
Brownian motion
Newtonian force
drag force
Particle in fluid
Solution is of exponential decay
Damping provided by water
Particle in ideal trap
Solution is of simple harmonic motion
Spring constant
Trapped particle in fluid
Solution is of damped simple harmonic motion
The whole picture
Time averaged effect is 0
Stochastic events introduce fluctuations in the particle’s position
Add in the effect of Brownian motion
Trap dynamics
Look at the movement of the particle in x and y
Power spectrum
Trap strength
Fourier transform to get the power spectrum
Lorenzian
Real data
Coming next
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Exam question?
In groups of 3 or 4, create two exam questions, one long, (6 marks), one short (3 marks).
5mins
Collecting data
How can we collect this data?
Moving 100s nm at a few kHz!!!
3 options
Option 1 - Camera
Camera placed in the image plane ofthe objective lens.
Uses the light fromthe illumination source.
Fast shutter speed to take clean image of particle.
Option 1 - Camera
Advantages
• Easy to use• Visual image of particle• Multiple particles
Disadvantages
• 2D measurement • Bandwidth limitations <100Hz• Very slow compared to f0
• Require very fast shutter so need a sensitive camera
Option 2 - Quadrant Photodiode A
Quadrant photodiode placed in the image plane of the objective lens (exactly the same as the camera).
Uses the light fromthe illumination source.
Option 2 - Quadrant Photodiode A
Advantages
• Large bandwidth 100s kHz• Very fast compared to f0
Disadvantages
• Single particle• Low light level, so small signal• 2D measurement
Option 3 - Quadrant Photodiode B
Quadrant photodiode collects the laser light transmitted through the condenser lens.
Small changes in the transmitted and scattered light are measured.
Option 3 - Quadrant Photodiode B
Advantages
• Large bandwidth 100s kHz• Very fast compared to f0
• 3D measurement•High light level as collecting
laser light
Disadvantages
• Complex arrangement• Single particle
Moving particles and multiple particles
Some background optics
An angular shift in the object plane results in a lateral shift in the imageplane.
Collimated light is brought to a focus a distance f, from a lens of focal length f.
Object plane
Image plane
Some background optics
If the beam is not collimated there isa shift in the axial position of the focus.
Moving objects around
f f f f f’ f’
Relay lenses
Beam steeringmirror
Angular deflection at mirror gives lateral shift of trap
Diffractive optics
Placing a diffractive optical element in theobject plane can generate a number of focused spots.
Diffraction grating
Spatial Light ModulatorsCalculated pattern
Video signal
Incoming beam
reflected/diffractedbeam
SLM
optical addressing
• Spatial light modulator = computer-controlled hologram– Liquid crystal (introduce phase or amplitude modulation)– Optically addressed SLMs convert intensity pattern to phase– diffraction efficiency >50%– >VGA resolution
Holograms at work
split the beam focus the beam
transform the beam combinations of the above
Whole beam path
SLM
beam-steerin
g mirror mirror imaged on
microscope entrance pupil
microscope objective
SLM imaged on beam-steering mirror
also: plane waves conserved
Holographic optical tweezers can do (just
about) anything!Hologram
Incident beam
Diffracted beams
Curtis et al. Opt. Commun. 207, 169 (2002)
• Holographic beam generation can create– multiple beams– modified beams– focussed beams– or all these at the
same time• REAL TIME control
of the beams
Dynamic multiple traps
Eriksen et al. Opt. Exp. 10, 597 (2002)
• Use spatial light modulator to create multiple traps– Lateral
displacement– Axial
displacement• Update trap positions
– Video frame rate
Rotating cube
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Exam question?
In groups of 3 or 4, create two exam questions, one long, (6 marks), one short (3 marks).
5mins
Applications of optical tweezers
Bio-applications
The size of particles that can be trapped is ~0.1m to 10’s m
Approximately the same size asmany biological specimen.
e.g. Blood cells, stem cells, DNA molecules
Either trapped directly, or beads used as handles to reduce optical damage.
Ashkin et al. Nature. 330, 768 (1987)
Block et al. Nature. 338, 514 (1989)
Measuring force/motion
Molloy et al. Biophys J. 68, S298 (1995)
biologicalobject
trapped bead
quadrant detector
imaging lens
• Image trapped bead (handle) onto quadrant detector
• Measure movement of shadow– nm accuracy!– kHz response
• Adjust trap to maintain position gives measurement of force– sub-pN accuracy!
e.g. Observation of myosin binding
• Handles attached to actin filament
• Intermittent binding to myosin suppresses thermal motion of beads due to stiff physical bond
e.g. Stretching/twisting of DNA
Perkins et al. Science. 264, 822 (1994)
Wang et al. Science. 282, 902 (1998)
• Attach handles to ends of DNA molecule
• Pull, let go and observe what happens!– understanding of
protein folding
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Work at Glasgow
5 microns5 microns
Jordan et al., J. Mod. Opt.,2004
• Permanent micro-structures
• Use SLM to create tweezers arrays
• Trap pseudo 2D crystals (≈100) (Curtis 2002)
• What happens when you turn the light off?– Fix structure in gel
• Permanent micro-structures
• Use SLM to create tweezers arrays
• Trap pseudo 2D crystals (≈100) (Curtis 2002)
• What happens when you turn the light off?– Fix structure in gel
Physical applications
Transfer of angular momentum
Angular momentum per photon = hbar
Angular momentum per photon = -hbar
If the particle Is birefringent it will absorb angular momentum and rotate.
Half-waveplate
Circularlypolarised light
Direction of propagation
Physical applications
Polarisation vectors rotatePolarisation vectors rotate(circular polarisation)(circular polarisation)
Spin angular momentumSpin angular momentum
Phase structure rotatesPhase structure rotates(helical phase fronts)(helical phase fronts)
Orbital angular Orbital angular momentummomentum
Padgett and Allen, Contemp. Phys. 41, 275 (2000)
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Absorption of orbital and spin angular momentum
Orbital AMOrbital AMSpinSpin
O’Neil et al. Phys. Rev. Lett. 053601 (2002)
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Microfluidic applications
Micro-machines driven by optical tweezers
optical micro-pump
Terray et al. Science. 296, 1841 (2002)
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• Translational (or rotational) control
• Fluid pumps• Optically driven stirring
Vortex Arrays
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Ladvac and Grier, Optics Express, 2004
Work at Glasgow
Optically driven pump using two counter rotating birefringent particles
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Flow
Flow meter
v = d/t
Turn laser on and off and measure particle displacement.
d
Work at Glasgow
Flow meter
Work at Glasgow
Flow meter results
A few of the (many) active groups
• World-wide– Grier et al. NY USA– Glückstad et al. Risø Denmark– Rubinstein-Dunlop et al. Queensland, Australia
• UK– Us! Glasgow– Dholakia et al. St Andrews– Molloy et al. National Institute for Medical Research, London
Conclusions
Trap dynamics and mechanisms
Positioning, manipulation and control
Bio, micro, physical applications
ConstantsN.A. = numerical aperturen = refractive index= anglea = radius of particle = wavelength of lightI0 = intensity nm = refractive index trapping mediumnp= refractive index particlem = np/nm (in the Fscatt, Fgrad equation)c = speed of lightm = mass (in the equation of motion)P = powerQ = trapping efficiencya = accelerationv = velocityx = positiont = timeT = temperaturekB = Boltzmann’s constantS = power spectrum = 6a= viscosity = trap strength
Exam question?
In groups of 3 or 4, create two exam questions, one long, (6 marks), one short (3 marks).
5mins