Gold nanorods as molecular contrast agents in photoacoustic imaging: the promises and the caveats\ud

Rod-shaped gold nanoparticles exhibit intense and narrow absorption peaks for light in the far-red and near-infrared wavelength regions, owing to the excitation of longitudinal plasmons. Light absorption is followed predominantly by non radiative de-excitation, and the released heat and subsequent temperature rise cause strong photoacoustic (optoacoustic) signals to be produced. This feature combined with the relative inertness of gold, and its favorable surface chemistry, which permits affinity biomolecule coupling, has seen gold nanorods (AuNR) attracting much attention as contrast agents and molecular probes for photoacoustic imaging. In this article we provide an short overview of the current status of the use of AuNR in molecular imaging using photoacoustics. We further examine the state of the art in various chemical, physical and biochemical phenomena that have implications for the future photoacoustic applications of these particles. We cover the route through fine-tuning of AuNR synthetic procedures, toxicity reduction by appropriate coatings, in vitro cellular interactions of AuNRs, attachment of targeting antibodies, in vivo fate of the particles and the effects of certain light interactions with the AuN

Twente Optical Perfusion Camera: system overview and performance for video rate laser Doppler perfusion imaging

We present the Twente Optical Perfusion Camera (TOPCam), a novel laser Doppler Perfusion Imager based on CMOS technology. The tissue under investigation is illuminated and the resulting dynamic speckle pattern is recorded with a high speed CMOS camera. Based on an overall analysis of the signal-to-noise ratio of CMOS cameras, we have selected the camera which best fits our requirements. We applied a pixel-by-pixel noise correction to minimize the influence of noise in the perfusion images. We can achieve a frame rate of 0.2 fps for a perfusion image of 128×128 pixels (imaged tissue area of 7×7 cm2) if the data is analyzed online. If the analysis of the data is performed offline, we can achieve a frame rate of 26 fps for a duration of 3.9 seconds. By reducing the imaging size to 128×16 pixels, this frame rate can be achieved for up to half a minute. We show the fast imaging capabilities of the system in order of increasing perfusion frame rate. First the increase of skin perfusion after application of capsicum cream, and the perfusion during an occlusion-reperfusion procedure at the fastest frame rate allowed with online analysis is shown. With the highest frame rate allowed with offline analysis, the skin perfusion revealing the heart beat and the perfusion during an occlusion-reperfusion procedure is presented. Hence we have achieved video rate laser Doppler perfusion imaging

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

Quantification of spatial intensity correlations and photodetector intensity fluctuations of coherent light reflected from turbid particle suspensions

We present a model for predicting the spatial intensity correlation function of dynamic speckle patterns formed by light backscattered from turbid suspensions, and an experimental validation of these predictions. The spatial correlation varies remarkably with multiple scattering. The provided computational scheme is a step towards correctly interpreting signals obtained from instruments based on the measurement of dynamic speckle patterns in the far field

Measurement of particle flux in a static matrix with suppressed influence of optical properties, using low coherence interferometry

Perfusion measurements using conventional laser Doppler techniques are affected by the variations in tissue optical properties. Differences in absorption and scattering will induce different path lengths and consequently will alter the probability that a Doppler shift will occur. In this study, the fraction of Doppler shifted photons and the Doppler broadening of a dynamic medium, are measured with a phase modulated low coherence Mach-Zehnder interferometer. Path length-resolved dynamic light scattering measurements are performed in various media having a constant concentration of dynamic particles inside a static matrix with different scattering properties and the results are compared with a conventional laser Doppler technique, with a simple model and with Monte Carlo simulations. We demonstrate that, for larger optical path lengths, the scattering coefficient of the static matrix in which the moving particles are embedded have a small to minimal effect on the measured fraction of Doppler shifted photons and on the measured average Doppler frequency of the Doppler shifted light. This approach has potential applications in measuring perfusion independent of the influence of optical properties in the static tissue matrix

The 'nanobig rods' class of gold nanorods: optimized dimensions for improved in vivo therapeutic and imaging efficacy

Currently, gold nanorods can be synthesized in a wide range of sizes. However, for intended biological applications gold nanorods with approximate dimensions 50 nm x 15 nm are used. We investigate by computer simulation the effect of particle dimensions on the optical and thermal properties in the context of the specific applications of photoacoustic imaging. In addition we discuss the influence of particle size in overcoming the following biophysical barriers when administrated in vivo: extravasation, avoidance of uptake by organs of the reticuloendothelial system, penetration through the interstitium, binding capability and uptake by the target cells. Although more complex biological influences can be introduced in future analysis, the present work illustrates that larger gold nanorods, designated by us as "nanobig rods", may perform relatively better at meeting the requirements for successful in vivo applications compared to their smaller counterparts which are conventionally used

Measurements of wavelength dependent scattering and backscattering coefficients by low-coherence spectroscopy

Quantitative measurements of scattering properties are invaluable for optical techniques in medicine. However, noninvasive, quantitative measurements of scattering properties over a large wavelength range remain challenging. We introduce low-coherence spectroscopy as a noninvasive method to locally and simultaneously measure scattering μs and backscattering μb coefficients from 480 to 700 nm with 8 nm spectral resolution. The method is tested on media with varying scattering properties (μs = 1 to 34 mm−1 and μb = 2.10−6 to 2.10−3 mm−1), containing different sized polystyrene spheres. The results are in excellent agreement with Mie theor

Evaluation of a multimode fiber optic low coherence interferometer for path length resolved Doppler measurements of diffuse light \ud

The performance of a graded index multimode fiber optic low coherence Mach-Zehnder interferometer with phase modulation is analyzed. Investigated aspects were its ability to measure path length distributions and to perform path length resolved Doppler measurements of multiple scattered photons in a turbid suspension of particles undergoing Brownian and translational motion. The path length resolution of this instrument is compared with a system using single mode fibers for illumination and detection. The optical path lengths are determined from the zero order moment of the phase modulation peak in the power spectrum. The weighted first moment, which is equal to the average Doppler shift, shows a linear response for different mean flow velocities within the physiological rang