RESUMEN
In this paper we present a general method for estimating rates of accidental coincidence between a pair of single photon detectors operated within their saturation regimes. By folding the effects of recovery time of both detectors and the detection circuit into an "effective duty cycle" we are able to accomodate complex recovery behaviour at high event rates. As an example, we provide a detailed high-level model for the behaviour of passively quenched avalanche photodiodes, and demonstrate effective background subtraction at rates commonly associated with detector saturation. We show that by post-processing using the updated model, we observe an improvement in polarization correlation visibility from 88.7% to 96.9% in our experimental dataset. This technique will be useful in improving the signal-to-noise ratio in applications which depend on coincidence measurements, especially in situations where rapid changes in flux may cause detector saturation.
RESUMEN
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.
Asunto(s)
ADN/química , Animales , Anisotropía , Calibración , Línea Celular Tumoral , Coloides/química , Diatomeas , Proteínas Fluorescentes Verdes/metabolismo , Microscopía Electrónica de Rastreo/métodos , Microscopía de Contraste de Fase/métodos , Microesferas , Nanotubos/química , Pinzas Ópticas , Óptica y Fotónica/métodos , Polisacáridos/química , Ratas , Dióxido de Silicio/química , Propiedades de SuperficieRESUMEN
We investigate the degree of control that can be exercised over an optically trapped microscopic non-spherical force probe. By position clamping translational and rotational modes in different ways, we are able to dramatically improve the position resolution of our probe with no reduction in sensitivity. We also demonstrate control over rotational-translational coupling, and exhibit a mechanism whereby the average centre of rotation of the probe can be displaced away from its centre.
Asunto(s)
Pinzas Ópticas , Óptica y Fotónica , Algoritmos , Diseño de Equipo , Rayos Láser , Micromanipulación/métodos , Modelos Estadísticos , Estrés Mecánico , Grabación en VideoRESUMEN
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.
RESUMEN
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.
RESUMEN
We report the implementation of a multitouch console for control of a holographic optical tweezers system. This innovative interface enables the independent but simultaneous interactive control of numerous optical traps by multiple users, overcoming the limitations of traditional interfaces and placing the full power of holographic optical tweezing into the operators' hands.