Anna Linnenberger

Increasing trap stiffness with position clamping in holographic optical tweezers

We present a holographic optical tweezers system capable of position clamping multiple particles. Moving an optical trap in response to the trapped objects motion is a powerful technique for optical control and force measurement. We have now realised this experimentally using a Boulder Nonlinear Systems Spatial Light Modulator (SLM) with a refresh rate of 203Hz. We obtain a reduction of 44% in the variance of the beads position, corresponding to an increase in effective trap stiffness of 77%. This reduction relies on the generation of holograms at high speed. We present software capable of calculating holograms in under 1ms using a graphics processor unit.

'Red tweezers': fast, customisable hologram generation for optical tweezers

Holographic Optical Tweezers (HOT) are a versatile way of manipulating microscopic particles in 3D. However, their ease of use has been hampered by the computational load of calculating the holograms, resulting in an unresponsive system. We present a program for generating these holograms on a consumer Graphics Processing Unit (GPU), coupled to an easy-to-use interface in LabVIEW (National Instruments). This enables a HOT system to be set up without writing any additional code, as well as providing a platform enabling the fast generation of other holograms. The GPU engine calculates holograms over 300 times faster than the same algorithm running on a quad core CPU. The hologram algorithm can be altered on-the-fly without recompiling the program, allowing it to be used to control Spatial Light Modulators in any situation where the hologram can be calculated in a single pass. The interface has also been rewritten to take advantage of new features in LabVIEW 2010. It is designed to be easily modified and extended to integrate with hardware other than our own.

A compact holographic optical tweezers instrument

Holographic optical tweezers have found many applications including the construction of complex micron-scale 3D structures and the control of tools and probes for position, force, and viscosity measurement. We have developed a compact, stable, holographic optical tweezers instrument which can be easily transported and is compatible with a wide range of microscopy techniques, making it a valuable tool for collaborative research. The instrument measures approximately 30×30×35 cm and is designed around a custom inverted microscope, incorporating a fibre laser operating at 1070 nm. We designed the control software to be easily accessible for the non-specialist, and have further improved its ease of use with a multi-touch iPad interface. A high-speed camera allows multiple trapped objects to be tracked simultaneously. We demonstrate that the compact instrument is stable to 0.5 nm for a 10 s measurement time by plotting the Allan variance of the measured position of a trapped 2 μm silica bead. We also present a range of objects that have been successfully manipulated.

Improving spatial light modulator performance through phase compensation

High-resolution, liquid-crystal spatial light modulators (SLMs) are being used as dynamic phase screens1,2 for testing optical systems and as optical wavefront compensators3,4 to dynamically correct distortions. An SLM provides hundreds of waves of adjustable phase modulation across the aperture of the device. Some of this phase adjustment can be used to compensate for distortions internal to the SLM such as backplane curvature. Because of modulo-2π operation, the dynamic range of the device is not significantly decreased by adding phase compensation, as long as the phase shift over the aperture is only a few waves. In this paper, we will discuss the techniques being used to determine the correct phase compensation for SLMs and how the compensation is being applied through the SLM control software.

Advances in optical phased array technology

Commercially available Liquid Crystal on Silicon (LCoS) Optical Phase Arrays (OPA) are capable of non-mechanically beamsteering up to ±3 degrees at 1550 nm. While the existing technology is useful for many applications such as laser communications and pulse-shaping, it is desirable to increase the steer angle and decrease the response time of the OPA. This was accomplished through a research effort funded by Langley Research Center at NASA. Under this research effort Boulder Nonlinear Systems (BNS) designed a new 1x12288 pixel OPA. In the new backplane design the pixel pitch was decreased from 1.8 um to 1.6 um, the backplane voltage was increased from 5 volts to 13 volts, and the aperture was increased from 7.4 x 6.0 mm to 19.66 x 19.66 mm. The OPA, when built with new liquid crystals and calibrated with new automated calibration procedures demonstrated a greater than 2x improvement in steer angle. The OPA that was tested, which was built for operation at 1550 nm, demonstrated the ability to steer to ±6.95 degrees. Additionally the relaxation time of the OPA was improved to 24.8 ms. This paper discusses the benefits of the new backplane design, the liquid crystal (LC) properties that are most desirable for beamsteering, the implementation of the automated calibration procedures, and the results.

Three dimensional live cell lithography

We investigate holographic optical trapping combined with step-and-repeat maskless projection stereolithography for fine control of 3D position of living cells within a 3D microstructured hydrogel. C2C12 myoblast cells were chosen as a demonstration platform since their development into multinucleated myotubes requires linear arrangements of myoblasts. C2C12 cells are positioned in the monomer solution with multiple optical traps at 1064 nm and then encapsulated by photopolymerization of monomer via projection of a 512x512 spatial light modulator illuminated at 405 nm. High 405 nm sensitivity and complete insensitivity to 1064 nm was enabled by a lithium acylphosphinate (LAP) salt photoinitiator. These wavelengths, in addition to brightfield imaging with a white light LED, could be simultaneously focused by a single oil immersion objective. Large lateral dimensions of the patterned gel/cell structure are achieved by x and y step-and-repeat process. Large thickness is achieved through multi-layer stereolithography, allowing fabrication of precisely-arranged 3D live cell scaffolds with micron-scale structure and millimeter dimensions. Cells are shown to retain viability after the trapping and encapsulation procedure.

Liquid-crystal-based hyperspectral image projector

A hyperspectral image projector (HIP) is introduced that is built with liquid crystal based spatial light modulators (SLM) as opposed to micromirror arrays. The use of an SLM as a broadband intensity modulator presents several benefits to this application. With slight modifications to the SLM design, SLMs can be built for a wide range of spectral regimes, ranging from the ultraviolet (UV) to the long-wavelength infrared (LWIR). SLMs can have a large pixel pitch, significantly reducing diffraction in the mid-wavelength infrared (MWIR) and LWIR. Liquid crystal based devices offer direct analog intensity modulation, thus eliminating flicker from time sequential drive schemes. SLMs allow for an on-axis configuration, enabling a simple and compact optical layout. The design of the HIP system is broken into two parts consisting of a spectral and spatial engine. In the spectral engine a diffraction grating is used to disperse a broadband source into spectral components, where an SLM modulates the relative intensity of the components to dynamically generate complex spectra. The recombined output is fed to the spatial engine which is used to construct two-dimensional scenes. The system is used to simulate a broad range of real world environments, and will be delivered to the National Institute of Standards and Technology as an enabling tool for the development of calibration standards and performance testing techniques for multispectral and hyperspectral imagers. The focus of this paper is on a visible-band HIP system; however, related work is presented with regard to SLM use in the MWIR and LWIR.

The Pocketscope: a spatial light modulator based epi-fluorescence microscope for optogenetics

Microscopy incorporating spatial light modulators (SLMs) enables three dimensional (3D) excitation and monitoring of the activity of neuronal ensembles, enabling studies of neuronal circuit activity both in vitro and in vivo. In this paper we present a portable (22 cm x 42.5 cm x 30 cm), SLM-based epi-fluorescence upright microscope (“Pocketscope”) that enables 3D calcium imaging and photoactivation of neurons in brain slices. Here we describe the implementation of the instrument; quantify the volume over which neural activity can be excited; and demonstrate the use of the system for mapping neural circuits in brain slices.

Three dimensional living neural networks

We investigate holographic optical tweezing combined with step-and-repeat maskless projection micro-stereolithography for fine control of 3D positioning of living cells within a 3D microstructured hydrogel grid. Samples were fabricated using three different cell lines; PC12, NT2/D1 and iPSC. PC12 cells are a rat cell line capable of differentiation into neuron-like cells NT2/D1 cells are a human cell line that exhibit biochemical and developmental properties similar to that of an early embryo and when exposed to retinoic acid the cells differentiate into human neurons useful for studies of human neurological disease. Finally induced pluripotent stem cells (iPSC) were utilized with the goal of future studies of neural networks fabricated from human iPSC derived neurons. Cells are positioned in the monomer solution with holographic optical tweezers at 1064 nm and then are encapsulated by photopolymerization of polyethylene glycol (PEG) hydrogels formed by thiol-ene photo-click chemistry via projection of a 512x512 spatial light modulator (SLM) illuminated at 405 nm. Fabricated samples are incubated in differentiation media such that cells cease to divide and begin to form axons or axon-like structures. By controlling the position of the cells within the encapsulating hydrogel structure the formation of the neural circuits is controlled. The samples fabricated with this system are a useful model for future studies of neural circuit formation, neurological disease, cellular communication, plasticity, and repair mechanisms.

Optical trapping for tissue scaffold fabrication

We investigate holographic optical trapping combined with step-and-repeat maskless projection stereolithography for fine control of 3D position of living cells within a 3D microstructured hydrogel. C2C12 myoblast cells were chosen as a demonstration platform because their development into multinucleated myotubes requires linear arrangements of myoblasts. C2C12 cells are positioned in the monomer solution with multiple optical traps at 1064 nm and then are encapsulated by photopolymerization of monomer via projection of a 512x512 spatial light modulator (SLM) illuminated at 405 nm. High 405 nm sensitivity and complete insensitivity to 1064 nm is enabled by a lithium acylphosphinate (LAP) salt photoinitiator. Use of a polyethylene glycol dimethacrylate (PEGDMA) based monomer is compared to that of polyethylene glycol (PEG) hydrogels formed by thiol-ene photo-click chemistry for patterning structures with cellular resolution, and for maintaining cell viability. Cells patterned in thiol-ene with RGD are shown to retain viability up to 4 days after the trapping and encapsulation procedure. Further, cells patterned in thiol-ene with RGD and a degradable ester link, are shown to fuse, indicating the initial stages of development of multi-nucleated cells.