Ultrasound-modulated optical tomography of absorbing objects buried in dense tissue-simulating turbid media

Continuous-wave ultrasonic modulation of scattered laser light was used to image objects buried in tissue-simulating turbid media. The buried object had an absorption coefficient greater than the background turbid medium. The ultrasonic wave that was focused into the turbid media modulated the laser light that passed through the ultrasonic field. The modulated laser light that was collected by a photomultiplier tube reflected the local mechanical and optical properties in the zone of ultrasonic modulation. Objects buried in the middle plane of 5-cm-thick dense turbid media were imaged with millimeter resolution through the scanning and detecting alterations of the ultrasound-modulated optical signal. The optical properties of the dense turbid media included an absorption coefficient of 0.1 cm(-1) and a reduced scattering coefficient of 10 cm(-1) and were comparable with those of biological tissues in the visible and near-IR ranges. The dependence of the ultrasound-modulated optical signal on the off-axis distance of the detector from the optic axis and the area of the detector was studied as well.     

Autocorrelation of scattered laser light for ultrasound-modulated optical tomography in dense turbid media

Based on measurement of the intensity autocorrelation function, a new method to determine the modulation depth of scattered laser light modulated by an ultrasonic wave in turbid media was applied to ultrasound-modulated optical tomography. Good signal-to-noise ratios and high sensitivities were demonstrated. Images of double optically absorbing objects buried in a highly optically scattering gel sample were obtained. The contrast was more than 10%, and the spatial resolution was approximately 2 mm.     

Precision of measurement of tissue optical properties with optical coherence tomography

Accurate and noninvasive measurement of tissue optical properties can be used for biomedical diagnostics and monitoring of tissue analytes. Noninvasive measurement of tissue optical properties (total attenuation and scattering coefficients, optical thickness, etc.) can be performed with the optical coherence tomography (OCT) technique. However, speckle noise substantially deteriorates the accuracy of the measurements with this technique. We studied suppression of speckle noise for accurate measurement of backscattering signal and scattering coefficient with the OCT technique. Our results demonstrate that the precision of measurement of backscattering signals with the OCT technique can be 0.2% for homogeneously scattering media and 0.7% for skin, if spatial averaging of speckle noise is applied. This averaging allows us to achieve the precision of tissue scattering coefficient measurements of approximately +/-0.8%. This precision can be further improved by a factor of 2-3, upon optimization of OCT operating parameters.     

In vivo demonstration of the ultrasound-modulated light technique

We present, to our knowledge for the first time, results of ultrasound-modulated light signals on living tissues. In particular, we analyze, both theoretically and experimentally, the effect of speckle fluctuations on the signal. We find that two different kinds of noise compete--shot noise and speckle noise--and are present at different levels in static phantoms and ex vivo tissue samples on the one hand and in dynamic phantoms and living tissues on the other hand.     

Fluorescence lifetime optical tomography in weakly scattering media in the presence of highly scattering inclusions

We consider the problem of fluorescence lifetime optical tomographic imaging in a weakly scattering medium in the presence of highly scattering inclusions. We suggest an approximation to the radiative transfer equation, which results from the assumption that the transport coefficient of the scattering media differs by an order of magnitude for weakly and highly scattering regions. The image reconstruction algorithm is based on the variational framework and employs angularly selective intensity measurements. We present numerical simulation of light scattering in a weakly scattering medium that embeds highly scattering objects. Our reconstruction algorithm is verified by recovering optical and fluorescent parameters from numerically simulated datasets.     

Computations of the acoustically induced phase shifts of optical paths in acoustophotonic imaging with photorefractive-based detection

Acoustophotonic imaging uses ultrasound-modulated scattered light to improve the quality of optical imaging in diffusive media. Experiments that use photorefractive-crystal-based detection have shown that there is a large dc shift in the acoustically modulated or ac optical signal, which could be utilized to further improve optical imaging resolution. We report that photon paths in a diffusive medium were generated by a Monte Carlo simulation, and the optical phase shifts of the various photons induced by the presence of a realistic focused ultrasound beam were calculated. Quantities that characterize the ac and dc signal components were evaluated by use of the calculated phase shifts. It was confirmed that the dc component dominates owing to coherent summation of the contributions from all the photons.     

Three-dimensional imaging of objects embedded in turbid media with fluorescence and Raman spectroscopy

We present a single-ended technique for three-dimensional imaging of objects embedded in a turbid medium by the use of time-resolved fluorescence emission or Raman scattering. The technique uses the earliest arriving photons, which we show are not sensitive to the relatively long fluorescence lifetime, and thus can be used to extract the desired spatial information accurately, even at a distance equivalent to 100 mean free paths. The results also demonstrate the feasibility and the potential of one's combining time-resolved optical tomography with fluorescence or Raman spectroscopy to localize and identify the embedded objects. This technique may be valuable for the diagnosis of disease in highly scattering human tissue because it can provide spatial and biochemical information about the composition of embedded lesions.     

Correlation transfer equation for multiply scattered light modulated by an ultrasonic pulse

We develop a temporal correlation transfer equation (CTE) and a Monte Carlo algorithm (MC) for multiply scattered light modulated by an ultrasonic pulse propagating in an optically scattering medium, where the ultrasound field can be nonuniform and the medium can have spatially heterogeneous distribution of optical parameters. The CTE and MC can be used to obtain the time-varying specific intensity and the spatial distribution of the time-dependent power spectral density, respectively, of ultrasound-modulated light. We expect the CTE and MC to be applicable for estimation of contrast and resolution in a wide spectrum of conditions in ultrasound-modulated optical tomography of soft biological tissues.     

Optical sectioning for optical scanning holography using phase-space filtering with Wigner distribution functions

We propose a novel optical sectioning method for optical scanning holography, which is performed in phase space by using Wigner distribution functions together with the fractional Fourier transform. The principle of phase-space optical sectioning for one-dimensional signals, such as slit objects, and two-dimensional signals, such as rectangular objects, is first discussed. Computer simulation results are then presented to substantiate the proposed idea.     

Three-dimensional localization and optical imaging of objects in turbid media with independent component analysis

A new approach for optical imaging and localization of objects in turbid media that makes use of the independent component analysis (ICA) from information theory is demonstrated. Experimental arrangement realizes a multisource illumination of a turbid medium with embedded objects and a multidetector acquisition of transmitted light on the medium boundary. The resulting spatial diversity and multiple angular observations provide robust data for three-dimensional localization and characterization of absorbing and scattering inhomogeneities embedded in a turbid medium. ICA of the perturbations in the spatial intensity distribution on the medium boundary sorts out the embedded objects, and their locations are obtained from Green's function analysis based on any appropriate light propagation model. Imaging experiments were carried out on two highly scattering samples of thickness approximately 50 times the transport mean-free path of the respective medium. One turbid medium had two embedded absorptive objects, and the other had four scattering objects. An independent component separation of the signal, in conjunction with diffusive photon migration theory, was used to locate the embedded inhomogeneities. In both cases, improved lateral and axial localizations of the objects over the result obtained by use of common photon migration reconstruction algorithms were achieved. The approach is applicable to different medium geometries, can be used with any suitable photon propagation model, and is amenable to near-real-time imaging applications.     

Image reconstruction by backprojection from frequency-domain optical measurements in highly scattering media

The reconstruction of the location and optical properties of objects in turbid media requires the solution of the inverse problem. Iterative solutions to this problem can require large amounts of computing time and may not converge to a unique solution. Instead, we propose a fast, simple method for approximately solving this problem in which calculated effective absorption and reduced scattering coefficients are backprojected to create an image of the objects. We reconstructed images of objects with centimeter dimensions embedded in a diffusive medium with optical characteristics similar to those of human tissue. Data were collected by a frequency-domain spectrometer operating at 120 MHz with a laser diode light source emitting at 793 nm. Intensity and phase of the incident photon density wave were collected from linear scans at different projection angles. Although the positions of the objects are correctly identified by the reconstructed images, the optical parameters of the objects are recovered only qualitatively.     

Theoretical and experimental studies of ultrasound-modulated optical tomography in biological tissue

Ultrasound-modulated optical tomography in biological tissue was studied both theoretically and experimentally. An ultrasonic beam was focused into biological tissue samples to modulate the laser light passing through the ultrasonic beam inside the tissue. The ultrasound-modulated laser light reflects the local optical and mechanical properties in the ultrasonic beam and permits tomographic imaging of biological tissues by scanning. Parallel detection of the speckle field formed by the transmitted laser light was implemented with the source-synchronous-illumination lock-in technique to improve the signal-to-noise ratio. Two-dimensional images of biological tissues were successfully obtained experimentally with a laser beam at either normal or oblique incidence, which showed that ultrasound-modulated optical tomography depends on diffuse light rather than on ballistic light. Monte Carlo simulations showed that the modulation depth decreased much more slowly than the diffuse transmittance, which indicated the possibility that even thicker biological tissues can be imaged with this technique.     

Two-dimensional inverse problem of diffusion tomography: the approach applicable for small inclusions

A method for solving the two-dimensional inverse problems of optical diffusion tomography is proposed. The method is especially designed for the imaging of small inclusions embedded in the backgrounds of strongly scattering media. Numerical simulations show that the results are stable with respect to external noise at the boundary of the sample. The location of an inclusion is obtained with an accuracy of the order of several photon transport mean-free paths in the medium in cases both with and without noise in the scattering data used for the solution of the inverse problem.     

Optical cross-sectional imaging with pulse ultrasound wave assistance.

We have developed an optical cross-sectional imaging method for turbid media with the aid of a pulse ultrasound wave. Observation of deep regions in turbid media, such as tissue samples, is difficult owing to the rapid dispersion of an incoming laser beam by scattering. A pulse ultrasound wave, which is less scattered in tissues, can indicate the measuring point on the basis of the change of the optical scattering properties in a localized region. A depth-resolving capability can be achieved from the time-dependent measurement of the scattered-light intensity as the pulse ultrasound wave propagates in the sample. We verified the method by observing absorptive objects embedded in silicone rubber and by obtaining the cross-sectional image of an absorbing object surrounded by a strong scattering medium.     

Motion-resolved imaging of moving objects embedded within scattering media by the use of time-gated speckle analysis

Spectral analysis of a time-evolving speckle pattern is used to provide motion-resolved detection of moving objects embedded within scattering media. Results show that the ability to detect small nonstationary scattering objects and to discriminate between objects moving at different rates is greatly enhanced.     

Methods for parallel-detection-based ultrasound-modulated optical tomography

The research reported here focuses on ultrasound-modulated optical tomography based on parallel speckle detection. Four methods were investigated for signal acquisition and analysis, in which laser speckle statistics was applied. The methods were compared with the previously used four-phase method in the imaging of all-biological-tissue samples, in which the buried objects were also biological tissues. The image quality obtained with these methods was comparable with that obtained with the four-phase method; in addition, these methods have advantages in reducing acquisition time and improving the signal-to-noise ratio.     

Recording invertebrate nerve activation with modulated light changes

Optical scattering techniques have the potential to provide noninvasive measurements of neural activity with good spatial and temporal resolution. We used the lobster nerve as a model system to investigate and record event-related optical signals with a modulated light source and heterodyne detection system. We observed changes in the transmitted birefringent light intensity, corresponding with electrophysiological measurements of the action potential. The photon delay was below the detection threshold, in part due to the small size of the nerve bundle. Our system allowed us to place an upper bound on the magnitude of the phase change of 0.01 degrees. The physiological stability of the preparation allows comprehensive characterization of biological and instrumentation noise sources for testing optical measurement systems.     

Eine Inkohärente Methode Aktiver Optischer Übertragung

We report the results of a simulation, using diffuse laser light and an optical bench, of our recent holographic interpretation of Labeyrie's speckle interferometry. Our approach applies to a wide class of multiple objects (such as close binary, or higher order, stars) viewed through a turbulent medium (such as the atmosphere, under all ‘seeing’ conditions). Our results should encourage astronomers to attempt to image those multiple star systems whose individual components would be resolvable in large telescopes were there no atmosphere to contend with, but whose details are always, or usually, obscured by atmospheric seeing. We find that we can get clear images of collections of small, isolated objects after processing only 20 speckle interferograms (we must emphasize, though, that the signal to noise ratio is high in our experiments i.e. each interferogram contains several hundred speckles). We find that our reconstructions of large, continuous objects are recognizable, although appreciably distorted. We attribute this to the virtual unavoidable non-linearities introduced by our processing methods, which are entirely optical and photographic. We suggest that improved results would be obtained by a suitable combination of optical and digital processing procedures. We point out that our results support those who hope to improve the performance of inferior optical instruments by using processing methods inspired by statistical concepts.     

Comparison of reconstruction algorithms for optical diffraction tomography

A recently developed inverse scattering algorithm [A. J. Devaney and M. Dennison, Inverse Probl., 19, 855 (2003) and M. Dennison and A. J. Devaney, Inverse Probl., 20, 1307 (2004)] is described and applied in a computer simulation study of optical diffraction tomography (ODT). The new algorithm is superior to standard ODT reconstruction algorithms, such as the filtered backpropagation algorithm, in applications employing a limited number of scattering experiments (the so-called limited-view case) and also in cases where multiple scattering occurs between the object being interrogated and the (known) background in which the object is embedded. The new algorithm is compared and contrasted with the filtered backpropagation algorithm in a computer simulation of ODT of weakly inhomogeneous cylindrical objects being interrogated in a limited number of scattering experiments employing incident plane waves. Our study has potential applications in biomedical imaging and tomographic microscopy.     

Laser excess noise reduction in optical phase-shift measurements

We describe an optical detection scheme that reduces laser excess noise in measurements of small optical phase shifts. The scheme improves the sensitivity of the balanced differential detection. Analysis of the scheme and comparison with the conventional detection are presented. The scheme is applied in electro-optic probing of electrical signals in integrated circuits, where the excess laser noise is reduced by approximately 20 dB.