Three-dimensional fluorescent microscopy via simultaneous illumination and detection at multiple planes.

The conventional optical microscope is an inherently two-dimensional (2D) imaging tool. The objective lens, eyepiece and image sensor are all designed to capture light emitted from a 2D 'object plane'. Existing technologies, such as confocal or light sheet fluorescence microscopy have to utilize mechanical scanning, a time-multiplexing process, to capture a 3D image. In this paper, we present a 3D optical microscopy method based upon simultaneously illuminating and detecting multiple focal planes. This is implemented by adding two diffractive optical elements to modify the illumination and detection optics. We demonstrate that the image quality of this technique is comparable to conventional light sheet fluorescent microscopy with the advantage of the simultaneous imaging of multiple axial planes and reduced number of scans required to image the whole sample volume

Adaptive Optimisation of Illumination Beam Profiles in Fluorescence Microscopy

Wide-field fluorescence microscope techniques such as single/selective plane illumination microscope (SPIM) are typically configured to image large regions of a sample at once. Here the illumination beam provides uniform excitation of several biological features across the region, `sliced' to a thickness of between 5-10 microns. In this paper we propose a simple alteration to the optical configuration of a SPIM by switching the light-sheet- forming cylindrical lens with a spatial light modulator. This has the potential to adaptively reconfigure the light sheet geometry to improve the optical sectioning of specific biological features, rather than the thicker sectioning of several features at once across a larger observation field-of-view. We present a prototype version of such a system, referred to as an Adaptive-SPIM (A-SPIM) system. We then suggest that the direct recording of illumination beam shapes within the working microscope system can better facilitate the analysis and subsequent re-configuration of the illumination beam to a specific geometry, and summarise the design and operation of a device that we have developed specifically for this purpose. We finally present reconstructed quantitative three dimensional flux maps of illumination beams from three microscope configurations taken using this miniature high-dynamic range beam profiling device, comparing the beam geometry of a regular SPIM system with our prototype A-SPIM system, and suggesting future improvements

Reflection mode photoacoustic measurement of speed of sound

We present a method to determine the speed of sound in tissue using a double-ring photoacoustic sensor working in reflection mode. This method uses the cross-correlation between the laser-induced ultrasound waves detected by two concentric ring shaped sensors, while a priori information about the depth-position of the photoacoustic source is not required. We demonstrate the concept by estimating the speed of sound in water as a function of temperature. Comparison of the estimated speed with values reported in literature shows an average systematic error of 0.1% and a standard deviation of 0.1%. Furthermore, we demonstrate that the method can be applied to layered media. The method has application in the correction of photoacoustic and ultrasound images afflicted by local speed variations in tissue. Additionally, the concept shows promise in monitoring temperature changes which are reflected in speed of sound changes in tissue.\ud \u

Tactile whole-field imaging sensor on photoelasticity

The paper describes a whole-field imaging sensor developed on the principles of photoelasticity. The sensor produces colored fringe patterns when load is applied on the contacting surface. These fringes can be analyzed using conventional photoelastic techniques, however, as the loading in the present case is not conventional some new strategies need to be devised to analyze the load imprint. The loading is unconventional in the sense that low modulus photoelastic material is deformed under vertical load in the direction of light travel to induce the photoelastic effect. The paper discusses the efficacy of both RGB calibration and phase shifting techniques in sensing applications. The characteristics of fringe patterns obtained under vertical and shear loads have been studied and the results obtained under these conditions are discussed with their limitations specifically when this is applied for sensing applications. Finally a case study has been conducted to analyze a foot image and conclusions drawn from this have been presented. Copyright © 2007 by ASME

Measurements of wall-shear stress fields on the piston crown in an IC engine flow using fluorescent labelled micro-pillar imaging

The measurement of wall shear-stress along the crown of a moving piston is accomplished with an array of hair-like, flexible micro-pillar sensors (MPS) in an internal combustion (IC) engine during intake flow conditions. The MPS are wall-clamped flexible micro-cylinders of 20micron diameter made of polydimethylsiloxane (PDMS) positioned on the surface of a transparent planar piston crown and protruding into the boundary layer flow above the piston at a height of 390micron. Their tips are labelled with fluorophores and are illuminated with a scanning laser-sheet system that follows the piston motion in synchronous manner. The flow-induced deflection is recorded through the transparent piston crown from below with a compact shaft-mounted high-speed camera recording the fluorescent re-emitted light from the pillar tips and using the fact of optical wave-guide features of the transparent micro-pillars. The experiments were performed in a transparent 4-valve engine setup (1.6 liter Prince 2, N13B16) during the intake phase. In order to improve the temporal resolution, the running speed of the engine was scaled down using water as working liquid. A static and dynamic sensor-calibration enabled the precise measurements of the wall-shear stress distribution with the sensor array. Due to spatial resolution limits of current available compact high-speed camera the recorded region along the piston was limited to 4x2 mm2 with 8x4 pillars in full view. The recordings at 300fps show the WSS fluctuations induced by the valve-jet / piston-wall interaction in the beginning of the intake with strong wall-normal motion forming critical points in the WSS field such as saddles, nodes and foci. Over the intake cycle > CA 80° these fluctuations die out and flow is transformed into wall-parallel coherent motion prescribed by the formation of the tumble

Solid state television camera system Patent

Solid state television camera system consisting of monolithic semiconductor mosaic sensor and molecular digital readout system