Background

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Importance of Project
* * *

Results
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SCREEN SCALING /CONVERSION FACTOR: The actual dimension of a grating line :

135mm
Grating on screen

White Lamp (20 W) Distance =30 m >30 m Average Speeds (m/s) 2.31 6.75 Maximum Speeds (m/s) (m/s) 11.45 22.22 Force (pN) 0.86 1.17

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Optical tweezers may provide the missing key for DNA base pair manipulation. manipulation. Lives can be saved if optical tweezers can safely perform precise movement of micro-particles. precise microFurther research can be conducted in molecular biology and other fields if tweezers can be demonstrated to trap small particles.

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11

10 7 6 5 4 1

Dimension of the line on screen: = 135mm Conversion factor for the dimensions measured on screen: k =

125,400m = 42.3m 2*300lines

= 0.3136m / mm

Previous Work
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Optical tweezers have been used effectively for almost twenty years [5]. years Three Skywalker team members participated in zoom lens design in Fall 2003. All members have laboratory experience and the required theoretical background. Reference [4] details popular physical layout and important issues. The current tweezers technology issues. can produce picoNewton (pN) forces on dielectric spheres.

For an object that appears as D(mm) on screen, the actual dimension will be d=kD.

A cold illumination (LED) assures larger distances of particle manipulation. (No extra heat, little thermal motion)

190mm
Ftrap = Fr = 6 rv (Stokes force for a constant speed motion in a fluid) Measured speed, v Ftrap
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LED Source (47mW) Distance 45 m Average speeds (m/s) 16.68 Maximum Speeds (m/s) (m/s) 17.3 Force (pN) 1.31

1 2 3 4 5

HeNe Laser Mirror Mirror L1, f =10mm L2, f = 76.8mm Iris Diaphragm L3, f =150mm Monochromatic Monitor CCD Camera Dichroic Mirror 100x Objective XYZ Translator Light Source Distances Between Parts 14 45 57 7 10 10 11 9 10 340 mm 86.8 mm 100 mm 216.8 mm 88.9 mm 90 mm

Screen size Figure 1: Screen Scaling

Approach and Methodology

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Concepts based on mathematical modeling of Gaussian beam optics [3]. Used ray matrices to trace beam spot size and radius of curvature through system of optics [2,3]. curvature

m i c r os p he r e s ' s pe e d s d i s tr i bu ti o n

Fr

* * *

Used CCD camera images to geometrically analyze positions of trapped particles. trapped Optical distances measured with millimeter ruler to ensure reproducibility of design. reproducibility Laser power and spot size along the path of the beam measured for estimation of the trapping force. for

7 8 9 10 11 12 13

m ic r o n s /s e c o

Development of Specifications

Trapped sphere

2 3
v = vmeas + vT vT =10 m , forLAMP; r
2 m , forLED

20

15

3.7 m;

10

10 3 Ns / m 2 ( water )

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18

Figure 2:Theory applied for calculating trapping force

Figure 3: speed for eighteen different trapped particles was measured

Overview
Project Goal: Goal: A working prototype of optical tweezers that can trap and manipulate micron-sized particles for manipulate micronapplications in biological research. Particles in the size range 3.2-4.4 microns were trapped range 3.2and moved in all three directions. The Following were Measured and Calculated: * The power at different points of the setup. * The force that the laser exerts on the particle. * The speeds with which the particles were moved with the laser beam. beam. Merits of the Design: The optical tweezer that has been designed can hold and manipulate micron- sized particles. micronThis provides the solution to difficulties faced in micro scale research. The sample illumination can be chosen between a light emitting diode and a white light source diode depending on how much power we want to use or how clear images we want. we

The spheres manipulation speed was provided by translating the stage while one particle was trapped. The speed was measured relative to other untrapped particles. For comparing the Brownian motion effect, two different light sources were used, and the thermal speed was measured for untrapped particles using screen distances. Without regarding the power used, the estimated forces, when moving the micro spheres with constant velocity, were in the range of pN.

OPTICAL INTERCEPTOR
Skywalkers Optical Tweezing Package
Optical Interceptor is a laser manipulator of micron-sized particles. Team micronSkywalkers is a research and development group that uses cutting edge technology to provide optical solutions to todays science.

The Influence of the Beam Size and Radius of Curvature upon the Trapping Force Waist Size (mm) Meas 0.9 Objective Aperture (mm) Proposed 7.3 Actual 5.99 M (magnification) Actual 7.7 Trial 3.9 f1 (mm) f2 (mm) d2 (mm) d1 (mm)

Actual 10

Trial 10

Actual 76.8

Trial 38.5

Actual 86.8

Trial 48.5

Proposed 200

Actual 340

For a trial 3.9 magnification and the values in red, the objective is not entirely filled, and no trapping effect was achieved. objective However the magnification was unchanged, moving Lens 3 for more than 1 cm, the trapping effect was lost. The radius of curvature was changed by moving Lens 3, and the best trapping effect was found to be where our initial best calculations had predicted.

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Power and Trapping Effect No. Power at the Pinhole (mW) 12 8 4 Trapping (Y/N) Y Y N Using polarizers (neutral density filters), we changed the power of the beam at the objective. No trapping was observed for powers less than 8 mW. Pinhole alignment was never achieved; however the pinhole position was used for measuring the correct distance to the objective. A diaphragmed Lens 1 ( =5 mm) and an iris (=20 mm) after Lens 3 were used for ( alignment. The meniscus of Lens 3 assured a more precise alignment compared to a plano-convex lens. plano-

1 2 3

TEAM SKYWALKERS MEMBERS

Specifications
The Optical Interceptor System is built with a useruserfriendly and easily upgradeable design. Laser Properties Type Power Wavelength Waist Size HeNe 25.5 mW 632.8 nm 0.9 mm Specification Can Translate Particle in X, Y, and Z Directions Trappable Particle Radius Recommended Translation Speed using LED Recommended Translation Speed using Incandescent Source Trapping Force Reproduction Cost of System Performance Yes * 3.2 - 4.4 m 6.9 m/s **

Emanuela Ene Theorist Maqsood Khandaker Marketing Advisor Benjamin Lee Design Coordinator Gim Khuan Ng System Operator Lakshmi Thamizhmani Technical Specialist Brady Whisenhunt Systems Analyst

Power Measurements Upon Exiting HeNe Laser Mirror 1 Mirror 2 Lens 1 Lens 2 Lens 3 Objective Power (mW) 25.5 22.0 16.0 15.0 15.0 14.0 12.0 Position At the Laser After lens 2 At the Minimum At the Objective At the Trap

Laser Spot Diameter Spot (mm) 0.71 7.53 < .025 7.2 0.0024 to 0.0048 Observations According to image analysis * Measured with caliper ** According to the pinhole aperture According to the geometrically illuminated area According to image analysis *

4.6 m/s ** 0.86 - 1.31 pN $5,522 ***

* Image analysis was performed using the Frame Grabber and MATLAB MATLAB software.**Measurements were standardized from image analysis. All values All were taken while using two ND filters.

Future Work

It is equipped with real-time image viewing and realcan instantaneously capture images with sizes of 45 m x 34 m on a 190 mm x 140 mm screen, giving an on-screen magnification of over on4000X.The image size at the focal plane was determined using a grating lens. The stage has knobs that allow for 250 m of translation (in the x, y, and z directions) per rotation with scale readings every 10 m.

* Z translation can be accomplished on a 10 m range, while X and Y translation is only limited by the stage size. ** Recommended speeds are approximately 40% of maximum speeds. Skywalker feels that this is a comfortable operating zone, but a skillful operator may attempt to achieve faster velocities at the risk of dropping particles. *** Cost of all parts and equipment except a computer.

References
Photonics Lab Website, http://www.ece.okstate.edu/photonicslab http://www.ece.okstate.edu/photonicslab E. Hecht, Optics, 2nd ed. Reading: Addison Wesley, 1989 Optics, J. T. Verdeyen, Laser Electronics, 3rd ed. New Jersey: Prentice Hall, 1995 Electronics, S. P. Smith, et al., Inexpensive optical tweezers for undergraduate laboratories, Am. J. Phys.67(1) , Jan.1999 Phys.67(1) [5] Physics Web, Optical Tweezers: The Next Generation, http://physicsweb.org/article/world/15/10/8 [1] [2] [3] [4]

The offsetting of focus can be set right by using a computer-assisted alignment system that allows for precise setting computerof focus and accurate error control. Power from the laser is not effectively transmitted to the objective because of the losses due to the mirrors. Using objective better mirrors and a higher power laser would be a possible solution to this problem. Note that better mirrors would solution have been used if square holders could have been obtained in tim e [1]. An infrared laser and a cuvette as sample holder will assure long living conditions for cell and microorganisms. long A better imaging system would help in precise manipulation of the beam and the system can be customized the according to the users needs. A larger monitor could be an initial improvement to the imaging system. (The system initial is equipped with a 9-inch monitor.) 9-

Weaknesses in the Design

Light Source Blue LED Incandescent Light

Screen Clarity Some Blurriness Very Bright

Fluid Motion Low High

The incandescent light allows better visibility, but the heat from from the lamp causes intense thermal particle motion. Although the LED gives less visibility, it reduces particle motion by reducing reducing the applied heat. The user will have the option to switch between the two light sources.

The designed optical tweezer could not trap and move E.coli bacteria from samples. It did not work for pseudomonas syringae, a plant pathogen. syringae, The optical apparatus is easily misaligned and calls in for a very skillful operator. very It is difficult to set the focus in the Z- direction. ZThe force exerted by the laser is not strong enough to trap particles that travel at greater speeds. particles The tweezer was unable to trap live particles and the solution dried up too soon resulting in the death of live cells.

In this picture several particles can be seen. They are the circular bright spots in the picture. A reference particle and a particle to be trapped have been outlined. The particle to be trapped will be held in the tweezing system and translated to the up and left, past the reference particle.

The particle to be trapped has now been caught inside the laser beam. The reflections of the beam from the particle inside the trap can easily be seen.

Now the trapped particle is being translated in the X and Y directions past the reference particle, which is moving only very slowly compared to the speed the trapped particle is moving past it.

Finally, the trapped particle is released in its new X and Y position to the left of the reference particle.

Reference Particle

Reference Particle

Trapped Particle After Translation

Trapped Particle Released

Particle to be Trapped Particle Trapped in Laser Beam

Reference Particle

Reference Particle