We study far from equilibrium systems through investigating how groups of bacteria behave when other bacteria are present in the system and when barriers are introduced. For these studies we use structured light to fabricate microscopic structures for optical trapping and cell studies using two photon photopolymerization process. Structured light is created using spatial light modulator and correct for wavefront distortions in-situ providing aberration corrected system. This system is used to enable production of simple holographic optical tweezers apparatus with as many as 50 individual foci to create complex 3D microstructures. These structures can induce the collective behaviour of bacteria.
We overview and further methods for using structured light to fabricate microscopic structures for optical trapping and cell study experiments through two-photon-photopolymerisation (2PP). We utilise a spatial light modulator (SLM) to correct wavefront distortions in-situ from gold nanoparticle reflections, which are shown to offer significant improvements for in-house nanofabrication. Aberration corrected 2PP offers a mechanism for using a simple, modified holographic optical tweezers apparatus to produce complex 3D micro-structures with as many as 50 independent foci. Custom-designed structures are used to induce and study the collective behaviour of bacteria allowing us to test otherwise simulated theories of cell dynamics.
Light can transfer both linear and angular momentum to matter. This constitutes the principle of optical tweezers developed by Arthur Ashkin and can be used for three dimensional trapping and manipulation of objects in a large variety of systems. The optical tweezers can be used in combination with three dimensional structures produced using two photon photopolymerisation process (2PP) in order to provide a source of all optically driven mircomachines that in turn are used for dynamic studies of biological and physical processes. They can also be used to construct heat engines.
Nanofabrication using two-photon-photopolymerision (2PP) can be used to create complex 3D structures with sub-diffraction-limited resolution for studying a range of microscopic systems. In this work we discuss how the addition of a spatial light modulator (SLM) can optimise the printing process through in-situ aberration correction and wavefront engineering. We show how digital holograms are used to control and fabricate complex patterns using various Gerchberg-Saxton algorithms. We demonstrate the necessity for aberration correction when printing using many independently controlled foci and fabricate devices that are used to study complex biological systems.
Piconewton scale forces can be measured with optical tweezers by monitoring the deflection of the trapping laser transmitted through a particle. This deflection is caused by a momentum transfer from the beam to the particle, measured as a change in voltage at a correctly placed position sensitive detector. To monitor these forces a conversion constant needs to be determined,which provides a mapping between units of voltage and Newtons. We propose, and experimentally verify, a new technique of detector calibration which outperforms previously developed methods. We also provide a quantitative comparison of different calibration techniques and the circumstances in which they are applicable.
We use dynamically controlled annular beam optical tweezers to orientate non-spherical swimming E.coli. Typically elliptically shaped particles in a Gaussian optical trap will align themselves with the direction of beam propagation. This orientation makes determining the dynamics of swimming particles difficult, as most optical tweezers systems are only able to capture information in the focal plane, but not the axial direction. We use simulations and experimental measurements to show that we are able to orientate motile E.coli from a vertical to horizontal position with a spatial light modulator in as little as one intermediate step without reduction in trap stiffness.
Using absolutely calibrated optical tweezers, we make quantitative measurements of the motility force of Escherichia coli (E. coli) by measuring the change in momentum of the deflected beam. By tracking the position of the particle, in addition to the optical force measurements, it should be possible to simultaneously calculate the motility force and drag. In a simple Gaussian beam optical trap away from the sample chamber E. coli tends to align and swim along the beam axis, which can make tracking the particle position and measuring the force difficult. We use a 3-D optical force detection system to measure the absolute force on the particle, allowing us to measure the motility force of E. coli in a simple Gaussian beam. By using a line-shaped trap, it is possible to align E. coli transverse to the beam axis, facilitating easy particle position measurement. The investigated methods are not specific to E. coli and could be applied to other motile organisms, the study of wall effects and bio-films.
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