David M. Carberry

Surface imaging using holographic optical tweezers

We present an imaging technique using an optically trapped cigar-shaped probe controlled using holographic optical tweezers. The probe is raster scanned over a surface, allowing an image to be taken in a manner analogous to scanning probe microscopy (SPM), with automatic closed loop feedback control provided by analysis of the probe position recorded using a high speed CMOS camera. The probe is held using two optical traps centred at least 10 µm from the ends, minimizing laser illumination of the tip, so reducing the chance of optical damage to delicate samples. The technique imparts less force on samples than contact SPM techniques, and allows highly curved and strongly scattering samples to be imaged, which present difficulties for imaging using photonic force microscopy. To calibrate our technique, we first image a known sample--the interface between two 8 µm polystyrene beads. We then demonstrate the advantages of this technique by imaging the surface of the soft alga Pseudopediastrum. The scattering force of our laser applied directly onto this sample is enough to remove it from the surface, but we can use our technique to image the algal surface with minimal disruption while it is alive, not adhered and in physiological conditions. The resolution is currently equivalent to confocal microscopy, but as our technique is not diffraction limited, there is scope for significant improvement by reducing the tip diameter and limiting the thermal motion of the probe.

An optically actuated surface scanning probe

We demonstrate the use of an extended, optically trapped probe that is capable of imaging surface topography with nanometre precision, whilst applying ultra-low, femto-Newton sized forces. This degree of precision and sensitivity is acquired through three distinct strategies. First, the probe itself is shaped in such a way as to soften the trap along the sensing axis and stiffen it in transverse directions. Next, these characteristics are enhanced by selectively position clamping independent motions of the probe. Finally, force clamping is used to refine the surface contact response. Detailed analyses are presented for each of these mechanisms. To test our sensor, we scan it laterally over a calibration sample consisting of a series of graduated steps, and demonstrate a height resolution of ∼ 11 nm. Using equipartition theory, we estimate that an average force of only ∼ 140 fN is exerted on the sample during the scan, making this technique ideal for the investigation of delicate biological samples.

Touching the microworld with force-feedback optical tweezers

Optical tweezers are a powerful tool for micromanipulation and measurement of picoNewton sized forces. However, conventional interfaces present difficulties as the user cannot feel the forces involved. We present an interface to optical tweezers, based around a low-cost commercial force feedback device. The different dynamics of the micro-world make intuitive force feedback a challenge. We propose a coupling method using an existing optical tweezers system and discuss stability and transparency. Our system allows the user to perceive real Brownian motion and viscosity, as well as forces exerted during manipulation of objects by a trapped bead.

Assembly and force measurement with SPM-like probes in holographic optical tweezers

In this paper we demonstrate the optical assembly and control of scanning probe microscopy (SPM)-like probes, using holographic optical tweezers. The probes are formed from cadmium sulphide rods and silica microspheres, the latter providing explicit trapping handles. Calibration of the trap stiffness allows us to use a precise measure of probe displacement to calculate the applied forces. We demonstrate that the optically controlled probe can exert a force in excess of 60 pN, over an area of 1×10−13 m2, with a force sensitivity of 50 fN. We believe that probes similar to the ones presented here will have applications as nanotools in probing laser-sensitive cells/materials.

Holographic assembly workstation for optical manipulation

We report a holographic assembler workstation for optical trapping and micro-manipulation. The workstation is based on a titanium sapphire laser, making it particularly suited for biomaterials and incorporates a choice of user interfaces for different applications. The system is designed around a commercial inverted microscope and is configured such that it can be easily used by the non-specialist. We demonstrate the bio-capabilities of our system by manipulating a group of yeast cells, a single red blood cell and a single cell of the green algae colony Volvox.

'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.

iTweezers: optical micromanipulation controlled by an Apple iPad

The 3D interactive manipulation of multiple particles with holographic optical tweezers is often hampered by the control system. We use a multi-touch interface implemented on an Apple iPad to overcome many of the limitations of mouse-based control, and demonstrate an elegant and intuitive interface to multi-particle manipulation. This interface connects to the tweezers system hardware over a wireless network, allowing it to function as a remote monitor and control device.

Measuring Nanoscale Forces with Living Probes

Optical trapping techniques have been used to investigate fundamental biological processes ranging from the identification of the processive mechanisms of kinesin and myosin to understanding the mechanics of DNA. To date, these investigations have relied almost exclusively on the use of isotropic probes based on colloidal microspheres. However, there are many potential advantages in utilizing more complex probe morphologies: use of multiple trapping points enables control of the interaction volume; increasing the distance between the optical trap and the sample minimizes photodamage in sensitive biological materials; and geometric anisotropy introduces the potential for asymmetric surface chemistry and multifunctional probes. Here we demonstrate that living cells of the freshwater diatom Nitzschia subacicularis Hustedt can be exploited as advanced probes for holographic optical tweezing applications. We characterize the optical and material properties associated with the high shape anisotropy of the silica frustule, examine the trapping behavior of the living algal cells, and demonstrate how the diatoms can be calibrated for use as force sensors and as force probes in the presence of rat B-cell hybridoma (11B11) cells.

Force sensing with a shaped dielectric micro-tool

We analyse the thermal motion of a holographically trapped non-spherical force probe, capable of interrogating arbitrary samples with nanometer resolution. High speed video stereo-microscopy is used to track the translational and rotational coordinates of the micro-tool in three dimensions, and the complete 6 × 6 stiffness matrix for the system is determined using equipartition theorem. The Brownian motion of the extended structure is described in terms of a continuous distribution of thermal ellipsoids. A centre of optical stress, at which rotational and translational motion is uncoupled, is observed and controlled. Once calibrated, the micro-tool is deployed in two modes of operation: as a force sensor with <150 femto-Newton sensitivity, and in a novel form of photonic force microscopy.

Calibration of optically trapped nanotools

Holographically trapped nanotools can be used in a novel form of force microscopy. By measuring the displacement of the tool in the optical traps, the contact force experienced by the probe can be inferred. In the following paper we experimentally demonstrate the calibration of such a device and show that its behaviour is independent of small changes in the relative position of the optical traps. Furthermore, we explore more general aspects of the thermal motion of the tool.