How does speckle contrast K, measured at camera exposures T around 10 ms, give us information about temporal autocorrelation of the speckle pattern with time constants τ < 1 ms, corresponding to Doppler shifts in the KHz range? We explore the implications of this question and show that for any particular assumed temporal speckle autocorrelation function, K measured at T >> τ accurately measures τ, but that K measurements at T < τ are required in order to determine the actual shape of the autocorrelation function. Determining the shape of the autocorrelation function is important if we wish to distinguish between different types of flow or movement in tissue, for example distinguishing Brownian motion or the randomly-oriented flows in capillary networks from more ordered flow in resolvable vessels.
The retina/choroid structure is an example of a complex biological target featuring highly perfused tissues and vessel flows both near the surface and at some depth. Laser speckle imaging can be used to image blood flows but static scattering paths present a problem for extracting quantifiable data. The speckle contrast is artificially increased by any residual specular reflection and light paths where no moving scatterers are encountered. Here we present results from phantom experiments demonstrating that the static and dynamic contributions to laser speckle contrast can be separated when camera exposures of varying duration are used. The stationary contrast parameter follows the thickness and strength of the overlying scatterer while the dynamic proportion of the scatter resulting from vessel flows and Brownian motion is unchanged. The importance of separating the two scatter components is illustrated by in vivo measurements from a scarred human retina, where the effect of the un-perfused scar tissue can be decoupled from the dynamic speckle from the intact tissue beneath it.
Measurements of flow in retinal vessels is presented and compared with in vitro measurements on whole blood in
capillaries ranging from 75 to 200μm diameter. The viewing angle of the capillaries and their range of size allows size-dependent
effects to be investigated when estimating flow within actual vessels.
Retinal measurements show a pulse effect. When this is removed by synchronisation, multi-exposure measurements
show different spectral signature from single speed scatterers. Multi-exposure measurements of the retina demonstrate a
varying contribution of stationary scatter across the field. Unlike scattering in dermal tissue, photons in retinal vessels
must return by multiple scatter from moving Red Blood Cells, whose motion is directed. Speckle estimates of flow in
retinal vessels are therefore possible.
Recent success in reconciling laser Doppler and speckle measurements of dermal perfusion by the use of multi-exposure
speckle has prompted an investigation of speckle effects arising from directed blood flow which might be expected in the
small blood vessels of the eye.
Unlike dermal scatter, the blood in retinal vessels is surrounded by few small and stationary scatterers able to assist the
return of light energy by large-angle scatter. Returning light is expected to come from multiple small angle scatter from
the large red blood cells which dominate the fluid.
This work compares speckle measurements on highly scattering skin, with measurements on flow in a retinal phantom
consisting of a glass capillary which is itself immersed in an index matching fluid to provide a flat air-phantom interface.
Brownian motion dominated measurements when small easily levitated scatters were used, and flow was undetectable.
With whole-blood, Brownian motion was small and directed flows in the expected region of tens of mm/s were
detectable. The nominal flow speed relates to the known pump rate; within the capillary the flow will have a profile
reducing toward the walls.
The pulsatile effects on laser speckle contrast in the retina are discussed with preliminary multi-exposure measurements
on retinal vessels using a fundus camera. Differences between the multiple exposure curves and power spectra of
perfused tissue and ordered flow are discussed.
Laser speckle and laser Doppler perfusion measurements apply different analyses to the same physical phenomenon and
so should produce the same results. However, there is some evidence that laser Doppler can measure perfusion at greater
depths than laser speckle. Using phantom measurements and comparison to spatially modulated imaging, we show why
this might be the case.
Various implementations of imaging laser Doppler and speckle systems have different optical setups, producing different
effective distances between the illumination and detector points on the surface of the tissue. Separating the effective
source and detector regions in tissue measurements biases the measurements towards deeper tissues, and when the
effective source and detector regions coincide, the measurement is biased towards surface tissues. Probe-based or
scanning laser Doppler systems with point illumination can separate the source and detector regions to interrogate deeper
tissues, while whole-field imaging laser Doppler systems and laser speckle contrast systems have broad illumination
covering the measurement areas. The volume of tissue informing a measurement at any point in a whole-field system,
and hence the depth sensitivity, is determined by the optical properties of the tissue at the working wavelength.
Variations in skin perfusion are easily detected by laser speckle contrast maps, but a robust interpretation of the information has been lacking. We show that multiple-exposure laser speckle methods produce the same spectral information as laser Doppler methods when applied to targets with embedded moving scatterers. This enables laser speckle measurements to be interpreted more quantitatively. We do this by using computer simulation of speckle data, and by experimental measurements on Brownian motion and skin perfusion using a laser Doppler system and a multiple-exposure laser speckle system. The power spectral density measurements of the light fluctuations derived using both techniques are exactly equivalent. Dermal perfusion can therefore be measured by laser Doppler or laser speckle contrast methods. In particular, multiexposure laser speckle can be rapidly processed to generate a full-field map of the perfusion index proportional to the concentration and mean velocity of red blood cells.
Laser speckle contrast measurements provide effectively instantaneous maps of dermal perfusion, using easily obtainable
hardware, but such maps are qualitative. Clinical applications of these techniques require a good theoretical and
experimental foundation of understanding before relating them to a physiologically significant, quantitative perfusion
value.
We have confirmed that multiple-exposure laser speckle methods produce the same spectral information as laser Doppler
measurements when applied to targets such as human tissue with embedded moving scatterers. This confirmation is
based on both computer simulation of laser speckle data and experimental measurements on Brownian motion and skin
perfusion using a laser Doppler system and a multiple-exposure laser speckle system. The Power Spectral Density (PSD)
measurements of the light fluctuations derived using both techniques are equivalent. Dermal perfusion images can
therefore be measured in exactly equivalent terms by either laser speckle contrast or more laborious scanning laser
Doppler methods.
Most analyses relating laser speckle contrast to perfusion depend on assuming a particular temporal autocorrelation
function for the light intensity fluctuations in biospeckle. Using multiple-exposure laser speckle allows the
autocorrelation function to be measured rather than assumed. Measured autocorrelation functions and their related power
spectra for dermal perfusion are presented, including measurements under arterial occlusion to investigate a 'biological
zero': the speckle blur relating to the remaining movement of tissue constituents when there is no net blood flow.
Laser speckle contrast techniques have been increasingly applied to dermal perfusion measurements over the past few
years. The interpretation of laser speckle contrast and its conversion to a physiologically-defined perfusion parameter
related to that found from Doppler measurements is becoming clearer. Speckle contrast-based techniques provide both
quantified perfusion images and a time-series record of perfusion.
We use the image resolution available in speckle measurements to investigate spatial resolution which can be expected in
tissue; in particular to reconcile speckle measurements with the large point-to-point variations reported from fibre
Doppler probes. In vitro models show the extent of spatial blurring likely to be encountered in speckle measurements at
different depths.
Perfusion responses related to vascular challenges could have medical relevance. We find a small pulse-related signal in
dermal speckle data. By identifying pulses in a temporal record using a matched filter, we find statistical average pulse
shapes for several different subjects, allowing comparison of pulsatile flow profiles between them. The profiles
measured by this technique are repeatable on the same subject, and vary between subjects. At some body sites, notably
near arterioles, the response obviously relates to gross tissue motion, but at others the signature is of dermal origin. It is
not yet clear whether it relates to actual capillary flow variation or distortion of the scattering tissue in response to
changes in the driving pressure.
Progress in laser and camera technology has simplified the acquisition of laser speckle images relating to dermal blood
flow. Using speckle contrast measurements over 5 decades of exposure time, we show that a temporal autocorrelation
function, and hence spectral information and a perfusion index precisely equivalent to that produced in Doppler methods,
can be derived from speckle measurements. The autocorrelation data are well approximated by a simple but nonexponential
function which is parametric in a characteristic time τc. We suggest that the perfusion index could be found
simply by determining τc from a small number of speckle measurements at appropriate exposures. This is illustrated by
measurement of perfusion recovery following an induced change in perfusion.
The fabrication of a designed arrangement of matter at the nano-scale level is a central goal of contemporary engineering endeavours. Silver nanoparticles synthesized by a laser ablation method in pure water are able to produce the aggregates, agglomerates and crystals due to condensed matter physics and chemistry. The stability of agglomerates and crystals is variable, depending on the composition of ensembles dominated by Ag2O and Ag respectively. The paper will present some fascinating nanostructures, such as films and vesicles.
An integrated oxygen control system has been explored by using bi-morph actuators based on polypyrrole conducting polymers (CP). The practical focus was control of a fruit storage atmosphere at 5% oxygen. An oxygen sensor and a simple actuator-valve assembly were integrated to limit the influx of atmospheric oxygen to a small chamber representing a fruit storage package. Electrochemical oxygen sensors such as zinc-air cells with output potentials up to one volt appear to be well suited for this task as they are able to drive trilayer strips based on CPs that actuate over a similar voltage range. The bimorph actuator used gave reliable and repeatable mechanical behaviour for about 50 hours.
In one sensor-actuator system trialled the output from a lead-oxygen sensor was electronically augmented to drive the actuator-valve. A set oxygen concentration of 5% was readily maintained with response times of tens of seconds. In the other system the voltage output from a zinc-air cell was used directly to control a very light actuator-valve that limited the free diffusion of air into the test chamber. Control of the oxygen concentration was achieved but at lower rates of oxygen flux and with longer response times.
Notable recent developments toward the realization of electronic nanocomputers have assembled logic circuits from semiconductor nanowires and individual carbon nanotube molecules. In spite of the broadly based and encouraging recent progress, a set of technical challenges still must be overcome to make a robust, commercially viable computer integrated on the molecular scale. The assembly of colloidal particles under an electric field offers many opportunities for the fabrication of ordered arrays, nanostructured films and microwires. We describe a method for the fabrication of gold nano/microstructures such as wires and dendrites on a lithographically patterned aluminium electrode with electric-field-induced assembly. The simple fabrication process will make these structures suitable for the miniaturisation of electronic circuits that can find application in sensors, actuators, and lab-on-a-chip devices. Our approach to electric-field-mediated fabrication exposes colloidal gold particles to the high electric field that can be generated between electrodes only 200 mm apart. We introduce an electric field of 100 Hz to 10 MHz by application of an alternating voltage of 5 to 10 V to the lithographically patterned microelectrodes. A suspension of gold nanoparticles of diameter 2.5 nm is added. We observe three types of fabrication, represented by three zones due to the different dielectophoretic force and convection effects. Some fibres grow through the liquid from one electrode toward the other, as could be seen in-situ by inverse optical microscopy. Dielectrophoretic-force-mediated fabrication, which is very flexible depending on the magnitudes of electric-field strength and frequency applied, has produced a notable advance in making mechanically flexible nano/microelectronic devices and led to a new understanding of the factors controlling the growth of nano/microstructures. When drops of suspension are patterned on the faces of components, three-dimensional structures can be generated. This type of system indicates how functional, self-assembling nano/microelectronic systems may be made. It provides a faster way of making devices, and the process can be very economical.
A system for detection of volatile compounds has been developed based on the concept of an electronic nose. The detection mechanism relies on the change in electrical resistance that occurs when a conducting polymer sensing element is exposed to the gaseous sample. An array of such sensors in conjunction with pattern recognition data analysis are used to identify and quantify the compounds of interest.
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