The effect of a 1070-nm continuous and pulsed wave ytterbium fiber laser on the growth of Saccharomyces cerevisiae single cells is investigated over a time span of 4 to 5 h. The cells are subjected to optical traps consisting of two counterpropagating plane wave beams with a uniform flux along the x, y axis. Even at the lowest continuous power investigated-i.e., 0.7 mW-the growth of S. cerevisiae cell clusters is markedly inhibited. The minimum power required to successfully trap single S. cerevisiae cells in three dimensions is estimated to be 3.5 mW. No threshold power for the photodamage, but instead a continuous response to the increased accumulated dose is found in the regime investigated from 0.7 to 2.6 mW. Furthermore, by keeping the delivered dose constant and varying the exposure time and power-i.e. pulsing-we find that the growth of S. cerevisiae cells is increasingly inhibited with increasing power. These results indicate that growth of S. cerevisiae is dependent on both the power as well as the accumulated dose at 1070 nm.
We have studied the effect of a 1070 nm continuous wave Ytterbium fiber laser on exponentially growing
Saccharomyces cerevisiae yeast cells over a span of 4 hours. The cells were immobilized onto Concanavalin A covered
microscope slides and the growth was measured using the area increase of the cells in 2D. Using a continuous dual beam
plane wave with a uniform spatial intensity distribution, we found that a continuous radiant flux through a single cell as
low as 0.5 mW in 1.5 hours significantly changed the growth and division rate of S. cerevisiae. With the dual beam setup
used we were able to successfully manipulate single S. cerevisiae cells in 3 dimensions with a minimum flux thorough
the cell of 3.5 mW. In the regime investigated from 0.7 mW to 2.6 mW we found no threshold for the photo damage, but
rather a continuous response to the increased accumulated dose.
The generation of optical landscapes may be regarded as a communication system: A user sends information to a
designated region in space where the information is represented using optical parameters like intensity. Information
capacity has been used for understanding superresolution in optical systems and we adapt this concept to gain useful
insights for characterizing techniques for generating optical landscapes. In particular, we investigate the information
capacity of the generalized phase contrast method and computer-generated holography. We also consider the invariance
of information capacity and discuss its implications for the generation of dynamic optical landscapes.
Optical trapping by use of multiple counter-propagating beam traps has not been widely implemented outside optical engineering laboratories. One, if not the primary, reason for this is the relatively complex calibration procedures involved in connection with this optical geometry. In this talk, we present automated solutions to all the calibration issues, which in effect results in a turn-key counter-propagating multi-beam 3D trapping system. These results allow a wider audience to utilize counter-propagating beam trapping systems. The calibrated system can be used to independently manipulate a plurality of cells real-time in a large 3D working area. Optionally, the system can be extended to allow for use of various spectroscopic methods concurrently with optical manipulation/trapping.
The generalized phase contrast method (GPC) can produce a rich variety of optical landscapes from an incident flattop
beam. Here we show that the GPC can generate various intensity distributions directly from an incident Gaussian
illumination. This is illustrated by using GPC-based implementation of phase-only apertures that efficiently redirect the
available photons from an initial bell-shaped intensity distribution into desired configurations. GPC can reshape a
Gaussian beam into patterns having sharp intensity transitions and a flat phase profile in the bright regions with superior
energy efficiency over amplitude masks. Eliminating the initial Gaussian-to-flattop beamshaping requirement can be
beneficial for various applications employing GPC-based patterns.
We outline the specifications of a portable Bio-photonics Workstation we have developed that utilizes just a single spatial light modulator to generate an array of up to 100 reconfigurable laser-traps with adjustable power ratios making 3D real-time optical manipulation possible with the click of a laptop mouse. We employ a simple patented optical mapping approach from a fast spatial light modulator to obtain reconfigurable intensity patterns corresponding to two independently addressable regions relayed to the sample volume where the optical manipulation of a plurality of nano-featured micro-objects takes place. The stand-alone Biophotonics Workstation is currently being tested by external partners with micro-biologic and chemistry expertise.
We demonstrate how optical trapping and manipulation can be used to assemble microstructures. The microstructures we
show being automatically recognized and manipulated are produced using the two-photon polymerization (2PP)
technique with submicron resolution. In this work, we show identical shape-complementary puzzle pieces being
manipulated in a fluidic environment forming space-filling tessellations. By implementation of image analysis to detect
the puzzle pieces, we developed a system capable of assembling a puzzle with no user interaction required. This allows
for automatic gathering of sparsely scattered objects by optical trapping when combined with a computer controlled
motorized sample stage.
Three-dimensional light structures can be created by modulating the spatial phase and
polarization properties of an an expanded laser beam. A particularly promising technique is
the Generalized Phase Contrast (GPC) method invented and patented at Risø National
Laboratory. Based on the combination of programmable spatial light modulator devices and
an advanced graphical user-interface the GPC method enables real-time, interactive and
arbitrary control over the dynamics and geometry of synthesized light patterns. Recent
experiments have shown that GPC-driven micro-manipulation provides a unique technology
platform for fully user-guided assembly of a plurality of particles in a plane, control of
particle stacking along the beam axis, manipulation of multiple hollow beads, and the
organization of living cells into three-dimensional colloidal structures. Here we present
GPC-based optical micromanipulation in a microfluidic system where trapping experiments
are computer-automated and thereby capable of running with only limited supervision. The
system is able to dynamically detect living yeast cells using a computer-interfaced CCD
camera, and respond to this by instantly creating traps at positions of the spotted cells
streaming at flow velocities that would be difficult for a human operator to handle.
Using a single phase-only spatial light modulator (SLM), we present a compact GPC-based optical trapping system for
interactively manipulating microscopic particles in three dimensions (3D) and in real-time. We employ only one GPC 4f
setup, which transforms 2D phase into intensity patterns, and utilize the SLM to form two phase-encoding regions
defined by two equally sized apertures - one centered at x = x0 and the other at x = -x0 (with the optical axis centered at
x = 0). Reconfigurable intensity patterns associated with the two independently addressable SLM-apertures are relayed
to the sample volume to form a dynamic array of counterpropagating-beam traps. We discuss the experimental
demonstrations showing 3D trapping of microparticles using the presented optical setup.
Helico-conical optical beams are a recently introduced class of beams that multiplicatively combine helical and conical phase fronts. Focusing these beams leads to a spiral intensity distribution at the focal plane of the lens. Further theoretical and experimental examination reveals interesting three-dimensional intensity patterns near the focal region, including a cork-screw structure around the optical axis. Variations on these light distributions based on the superposition of multiple helico-conical beams are also presented here. These beams are expected to yield interesting dynamics when applied to the optical trapping of microscopic particles, such as dielectric microspheres or even biological cells.
In the area of manipulating microscopic biological specimen, optical trapping has proven its worth. Still, many potential microbiological applications can benefit when the experiment is assisted by a computer and capable of running either with only limited supervision or full automation. Here we have implemented the Generalized Phase Contrast (GPC) method of optical trapping in a microfluidic system, and show how an experiment can be easily made to run autonomously, while the computer continuously adapts the light pattern to trap yeast cells passing through the trapping volume. The optical trapping takes place in a microfluidic system where two channels meet, allowing for separate injection of specimen and work media. Yeast cells are trapped near the surface of the microchannel at flow rates that give particle speeds of more than 50 μm/s. We demonstrate the ability of GPC-based traps to hold the cells in specific positions and observe the displacement of the cells from respective trap centers. Finally we exploit the speed of the GPC system by dynamically detect yeast cells using a CCD camera and immediately create traps at their positions at flow rates that exceed what a human operator would be able to handle. The optical system was found to be easily expanded and the attention could be kept on maintaining optimal conditions for the yeast.
The three-dimensional forces acting on a dielectric microparticle illuminated by two counterpropagating beams with variable intensity profiles are theoretically studied. Size-adjustable intensity profiles of the constituent beams are easily implemented using the generalized phase contrast (GPC) method. Our numerical calculations include the dependence of the axial and transverse trapping forces with location of the microsphere between the opposing beams. These numerical results show potential improvement in the large dynamic range for axial position control of microparticles in GPC-based counterpropagating-beam traps - achieved with the use of size-varied constituent tophat beams. We also assess the quality of the obtained trap potentials by calculating the axial and transverse stiffness associated with the optical traps.
We explore the Generalized Phase Contrast (GPC) approach for optical sorting in microfluidic systems. A microsystem is used in which two streams meet, interact and separate in an X-shaped channel. When the flow in the two arms of the X is balanced, the laminar flow that exists at very low Reynolds numbers ensures minimal stream blending and the fluid separates without mixing (i.e. diffusion is negligible). Optical forces due to an intensity pattern can be fashioned to induce a selective deflection of particles between the two streams. This method is known as optical fractionation (OF). In brief, OF uses the same mechanisms as optical tweezers to exert forces upon microscopic particles. OF has been shown to have an exponential size selectivity. This means that the interaction between the streams can be made to discriminate by particle size at a critical flow velocity. With correctly adjusted flow velocity, particles with a certain size will more often shift to the other stream than another particle size. One method for creating the light pattern is by interference of several beams that are variably attenuated using mechanical means. However, this approach offers low
optical efficiency and is not easily reconfigured. The GPC method offers a solution that gives the possibility to instantaneously reconfigure the intensity pattern by a method that is inherently computer-controllable. This enables one to rapidly test various intensity patterns to optimize sorting of particles.
We demonstrate the use of multiple optical traps for driving various microfabricated silica structures in liquid host medium. Multiple counterpropagating-beam traps are formed using a generalized phase contrast (GPC) -based optical trapping system. A combination of UV-lithography and reactive-ion etching (RIE) is employed to fabricate the microtools whose design includes having multiple appendages with rounded endings by which optical traps hold and actuate them. Experiments show the collective and user-coordinated utility of multiple beams for driving microstructured objects whose future integration may lead to optically controlled micromachineries.
Three-dimensional light structures can be created by modulating the spatial phase and polarization properties of the laser light. A particularly promising technique is the Generalized Phase Contrast (GPC) method invented and patented at Riso National Laboratory. Based on the combination of programmable spatial light modulator devices and an advanced graphical user-interface the GPC method enables real-time, interactive and arbitrary control over the dynamics and geometry of synthesized light patterns. Recent experiments have shown that GPC-driven micro-manipulation provides a unique technology platform for fully user-guided assembly of a plurality of particles in a plane, control of particle stacking along the beam axis, manipulation of multiple hollow beads, and the organization of living cells into three-dimensional colloidal structures. These demonstrations illustrate that GPC-driven micro-manipulation can be utilized not only for the improved synthesis of functional microstructures but also for non-contact and parallel actuation crucial for sophisticated opto- and micro-fluidic based lab-on-a-chip systems.
The generalized phase contrast (GPC) method is used to transform a collimated Gaussian beam into a manifold of counterpropagating-beam traps capable of real-time interactive manipulation of a plurality of particles in three dimensions. Contrary to high-NA based optical tweezers, GPC can operate with a centimeter long working distance, avoiding the use of objective lens immersion liquid and offers a wider manipulation region and a larger field of view than optical tweezers. As a consequence of the large working distance, simultaneous monitoring of the trapped particles in orthogonal observation planes can be easily integrated. Real-time 3D reconfigurable light patterns with sub-micron accuracy are obtained from a direct map of phase patterns addressed on phase-only spatial light modulator followed by a programmable parallel polarisation modulating device. A graphical user-interface enables interactive and arbitrary control over the dynamics and geometry of the real-time synthesized light patterns. Experimental demonstrations have shown that GPC-driven micro-manipulation can be used for fully user-guided assembly of particles in a volume, control of particle stacking along the optical axis, manipulation of multiple low-index particles, real-time sorting of inhomogeneous mixtures of micro-particles and the organization of living cells into 3D reconfigurable colloidal structures.
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