Image correction algorithm for functional three-dimensional diffuse optical tomography brain imaging

We outline a computationally efficient image correction algorithm, which we have applied to diffuse optical tomography (DOT) image time series derived from a magnetic resonance imaging (MRI)-based brain model. Results show that the algorithm increases spatial resolution, decreases spatial bias, and only modestly reduces temporal accuracy for noise levels typically seen in experiment, and produces results comparable to image reconstructions that incorporate information from MRI priors. We demonstrate that this algorithm has robust performance in the presence of noise, background heterogeneity, irregular external and internal boundaries, and error in the initial guess. However, the algorithm introduces artifacts when the absorption and scattering coefficients of the reference medium are overestimated--a situation that is easily avoided in practice. The considered algorithm offers a practical approach to improving the quality of images from time-series DOT, even without the use of MRI priors.     

Time-resolved imaging of solid phantoms for optical mammography

We have recorded time-resolved transillumination images of solid phantoms with objects embedded that differ in their scattering and absorption coefficients from those of the bulk material, simulating a compressed human breast with a tumor inside. Employing time-correlated single photon counting at rates of up to 1 MHz, we recorded distributions of times of flight of photons at 1369 scan positions within 2.5 min. Several quantities, such as fractional transmittance, first moments, Fourier amplitudes, phase shifts, and frequency-dependent effective transport scattering and absorption coefficients, have been derived from experimental data to form two-dimensional images. By recording such images at a selected total number of photons detected, we have determined the contrast and effective signal-to-noise ratio in each case.     

A new optical coefficient of biological tissue that determines the diffuse reflectance light distribution on the tissue surface

When a light beam is incident normally upon a tissue, the spatial distribution of diffuse reflectance on the tissue surface is affected by the optical properties of the tissue. The purpose of this paper is to find the relationship between the spatial distribution of diffuse reflectance on the tissue surface and optical properties of the tissue. The diffuse reflected light distribution is found to be determined mainly by a new optical coefficient μ_as. A qualitative relationship between μ_as and the diffuse reflected light distribution was developed by Monte Carlo simulations. Measurements of the diffuse reflected light distribution on solutions of Intralipid-10% with added ink confirm the Monte Carlo simulations. The new optical coefficient μ_as was measured from several tissue examples by the qualitative relationship. The value of μ_as was useful for determining the components of the tissue.     

Spatially resolved absolute diffuse reflectance measurements for noninvasive determination of the optical scattering and absorption coefficients of biological tissue

The absorption and transport scattering coefficients of biological tissues determine the radial dependence of the diffuse reflectance that is due to a point source. A system is described for making remote measurements of spatially resolved absolute diffuse reflectance and hence noninvasive, noncontact estimates of the tissue optical properties. The system incorporated a laser source and a CCD camera. Deflection of the incident beam into the camera allowed characterization of the source for absolute reflectance measurements. It is shown that an often used solution of the diffusion equation cannot be applied for these measurements. Instead, a neural network, trained on the results of Monte Carlo simulations, was used to estimate the absorption and scattering coefficients from the reflectance data. Tests on tissue-simulating phantoms with transport scattering coefficients between 0.5 and 2.0 mm(-1) and absorption coefficients between 0.002 and 0.1 mm(-1) showed the rms errors of this technique to be 2.6% for the transport scattering coefficient and 14% for the absorption coefficients. The optical properties of bovine muscle, adipose, and liver tissue, as well as chicken muscle (breast), were also measured ex vivo at 633 and 751 nm. For muscle tissue it was found that the Monte Carlo simulation did not agree with experimental measurements of reflectance at distances less than 2 mm from the incident beam.     

Spatial deconvolution technique to improve the accuracy of reconstructed three-dimensional diffuse optical tomographic images

A straightforward spatial deconvolution operation is presented that seeks to invert the information-blurring property of first-order perturbation algorithms for diffuse optical tomography (DOT) image reconstruction. The method that was developed to generate these deconvolving operators, or filters, was conceptually based on the frequency-encoding process used in magnetic resonance imaging. The computation of an image-correcting filter involves the solution of a large system of linear equations, in which known true distributions and the corresponding recovered distributions are compared. Conversely, application of a filter involves only a simple matrix multiplication. Simulation results show that application of this deconvolution operation to three-dimensional DOT images reconstructed by the solution of a first-order perturbation equation (Born approximation) can yield marked enhancement of image quality. In the examples considered, use of image-correcting filters produces obvious improvements in image quality, in terms of both location and mirco(a) of the inclusions. The displacements between the true and recovered locations of an inclusion's centroid location are as small as 1 mm, in an 8-cm-diameter medium with 1.5-cm-diameter inclusions, and the peak value of the recovered micro(a) for the inclusions deviates from the true value by as little as 5%.     

Design and implementation of dynamic near-infrared optical tomographic imaging instrumentation for simultaneous dual-breast measurements

Dynamic near-infrared optical tomographic measurement instrumentation capable of simultaneous bilateral breast imaging, having a capability of four source wavelengths and 32 source-detector fibers for each breast, is described. The system records dynamic optical data simultaneously from both breasts, while verifying proper optical fiber contact with the tissue through implementation of automatic schemes for evaluating data integrity. Factors influencing system complexity and performance are discussed, and experimental measurements are provided to demonstrate the repeatability of the instrumentation. Considerations in experimental design are presented, as well as techniques for avoiding undesirable measurement artifacts, given the high sensitivity and dynamic range (1:10(9)) of the system. We present exemplary clinical results comparing the measured physiologic response of a healthy individual and of a subject with breast cancer to a Valsalva maneuver.     

Multichannel time-resolved tissue oximeter for functional imaging of the brain

This paper reports the development and characterization of a novel multichannel time-resolved (TR) instrument for functional brain imaging studies. The instrument is based on picosecond diode lasers, fiber optics for light injection and delivery, a compact multianode photomultiplier, and a personal computer (PC) board for time-correlated single photon counting (TCSPC). The instrument has been characterized in terms of reproducibility among the nine sources and the 12 collection channels, linearity in the determination of optical properties (absorption and reduced scattering), and stability. Preliminary in vivo measurements were performed on volunteers to monitor the optical response to stimuli following a motor task (finger opposition, 5 Hz).     

Comparative evaluation of two simple diffuse reflectance models for biological tissue applications

We present a comparative evaluation of two simple diffuse reflectance models for biological tissue applications. One model is based on a widely accepted and used in biomedical optics implementation of diffusion theory, and the other one is based on a semiempirical approach derived from basic physical principles. We test the models on tissue phantoms and on human skin, utilizing a standard six-around-one optical fiber probe for light delivery and collection. We show that both models are suitable for use with an optical fiber probe and illustrate the potential, applicability, and validity range of the models.     

Depolarization and blurring of optical images by biological tissue

We present a study of the image blurring and depolarization resulting from the transmission of a narrow beam of light through a continuous random medium. We investigate the dependence of image quality degradation and of depolarization on optical thickness, correlation length of the inhomogeneities, and incident polarization state. This is done numerically with a Monte Carlo method based on a transport equation that takes into account polarization of light. We compare our results with those for transport in media with discrete spherical scatterers. We show that depolarization effects are different in these two models of biological tissue.     

Holographic digital Fourier microscopy for selective imaging of biological tissue

We present an application of digital Fourier holography for selective imaging of scatterers with different sizes in turbid media such as biological tissues. A combination of Fourier holography and high‐resolution digital recording, digital Fourier microscopy (DFM) permits crucial flexibility in applying filtering to highlight scatterers of interest in the tissue. The high‐resolution digital hologram is a result of the collation of Fourier holographic frames to form a large‐size composite hologram. It is expected that DFM has an improved signal‐to‐noise ratio as compared to conventional direct digital imaging, e.g., phase microscopy, as applied to imaging of small‐size objects. The demonstration of the Fourier filtering capacity of DFM using a biological phantom represents the main focus of this article. © 2005 Wiley Periodicals, Inc. Int J Imaging Syst Technol, 14, 253–258, 2004; Published online inWiley InterScience (www.interscience.wiley.com). DOI 10.1002/ima.20031     

Spectroscopic diffuse optical tomography for the quantitative assessment of hemoglobin concentration and oxygen saturation in breast tissue

Near-infrared (NIR) spectroscopic diffuse tomography has been used to map the hemoglobin concentration and the hemoglobin oxygen saturation quantitatively in tissuelike phantoms and to determine average values in vivo. A series of phantom calibrations were performed to achieve quantitatively accurate images of the absorption and the reduced scattering coefficients at multiple optical wavelengths. A least-squares fit was applied to absorption-coefficient images at multiple NIR wavelengths to obtain hemoglobin images of the concentration and the hemoglobin oxygen saturation. Objects of varying hemoglobin concentration and oxygen saturation within highly scattering media were localized and imaged to within 15% of their actual values. The average hemoglobin concentration and oxygen saturation of breast tissue was measured in vivo for two women volunteers. The potential application for the diagnosis of breast tumors is discussed.     

Hyperspectral diffuse reflectance imaging for rapid, noncontact measurement of the optical properties of turbid materials

We present a method and technique of using hyperspectral diffuse reflectance for rapid determination of the optical properties of turbid media. A hyperspectral imaging system in line scanning mode was used to acquire spatial diffuse reflectance profiles from liquid phantoms made up of absorbing dyes and fat emulsion scatterers over the spectral range of 450-1000 nm instantaneously. The hyperspectral reflectance data were analyzed by using a steady-state diffusion approximation model for semi-infinite homogeneous media. A calibration procedure was developed to compensate the nonuniform instrument response of the imaging system, and a curve-fitting algorithm was used to extract absorption and reduced scattering coefficients (mua and mus', respectively) for the phantoms in the wavelength range from 530 to 900 nm. The hyperspectral imaging system gave good measures of mua and mus' for the phantoms with average fitting errors of 12% and 7%, respectively. The hyperspectral imaging technique is fast, noncontact, and easy to use, which makes it especially suitable for measurement of the optical properties of turbid liquid and solid foods.     

Fast multispectral diffuse optical tomography system for in vivo three-dimensional imaging of seizure dynamics

We describe a multispectral continuous-wave diffuse optical tomography (DOT) system that can be used for in vivo three-dimensional (3-D) imaging of seizure dynamics. Fast 3-D data acquisition is realized through a time multiplexing approach based on a parallel lighting configuration--our system can achieve 0.12 ms per source per wavelength and up to a 14 Hz sampling rate for a full set of data for 3-D DOT image reconstruction. The system is validated using both static and dynamic tissue-like phantoms. An initial in vivo experiment using a rat model of seizure is also demonstrated.     

Effective scattering coefficient of the cerebral spinal fluid in adult head models for diffuse optical imaging

An efficient computation of the time-dependent forward solution for photon transport in a head model is a key capability for performing accurate inversion for functional diffuse optical imaging of the brain. The diffusion approximation to photon transport is much faster to simulate than the physically correct radiative transport equation (RTE); however, it is commonly assumed that scattering lengths must be much smaller than all system dimensions and all absorption lengths for the approximation to be accurate. Neither of these conditions is satisfied in the cerebrospinal fluid (CSF). Since line-of-sight distances in the CSF are small, of the order of a few millimeters, we explore the idea that the CSF scattering coefficient may be modeled by any value from zero up to the order of the typical inverse line-of-sight distance, or approximately 0.3 mm(-1), without significantly altering the calculated detector signals or the partial path lengths relevant for functional measurements. We demonstrate this in detail by using a Monte Carlo simulation of the RTE in a three-dimensional head model based on clinical magnetic resonance imaging data, with realistic optode geometries. Our findings lead us to expect that the diffusion approximation will be valid even in the presence of the CSF, with consequences for faster solution of the inverse problem.     

Determination of the optical coefficients of biological tissue by neural network

A system that incorporated a laser source and a CCD camera was used to measure spatially-resolved steady-state diffuse reflectance. Monte Carlo simulations and experiments in tissue phantoms were used to train a neural network that characterizes the reflectance data on a turbid medium. The neural network was used to extract the optical properties (scattering and absorption coefficients) of biological tissue. The accuracy of the neural network was investigated and validated. Tests on tissue-simulation phantoms showed the relative errors of this technique to be 3% for the reduced scattering coefficient and 9% for the absorption coefficients. The optical properties of human skin were also measured in vivo at 633 nm. For human skin tissue it was found that our results were in good agreement with their reference values.     

Fiducial markers for combined 3-dimensional mass spectrometric and optical tissue imaging

Mass spectrometric imaging (MSI) has become widely used in the analysis of a variety of biological surfaces. Biological samples are spatially, morphologically, and metabolically complex. Multimodal molecular imaging is an emerging approach that is capable of dealing with this complexity. In a multimodal approach, different imaging modalities can provide precise information about the local molecular composition of the surfaces. Images obtained by MSI can be coregistered with images obtained by other molecular imaging techniques such as microscopic images of fluorescent protein expression or histologically stained sections. In order to properly coregister images from different modalities, each tissue section must contain points of reference, which are visible in all data sets. Here, we report a newly developed coregistration technique using fiducial markers such as cresyl violet, Ponceau S, and bromophenol blue that possess a combination of optical and molecular properties that result in a clear mass spectrometric signature. We describe these fiducial markers and demonstrate an application that allows accurate coregistration and 3-dimensional reconstruction of serial histological and fluorescent microscopic images with MSI images of thin tissue sections from a breast tumor model.     

Time-resolved fluorescence polarization dynamics and optical imaging of Cytate: a prostate cancer receptor-targeted contrast agent

Cypate-octreote peptide analogue conjugate (Cytate) was investigated as a prostate cancer receptor-targeted contrast agent. The absorption and fluorescence spectra of Cytate were ranged in the near-infrared "tissue optical window." Time-resolved investigation of polarization-dependent fluorescence emitted from Cytate in solution as well as in cancerous and normal prostate tissues was conducted. Polarization preservation characteristics of Cytate in solution and tissues were studied. Fluorescence intensity emitted from the Cytate-stained cancerous prostate tissue was found to be much stronger than that from the Cytate-stained normal prostate tissue, indicating more Cytate uptake in the former tissue type. The polarization anisotropy of Cytate contained in cancerous prostate tissue was found to be larger than that in the normal prostate tissue, indicating a larger degree of polarization preservation in Cytate-stained cancerous tissue. The temporal profiles of fluorescence from Cytate solution and from Cytate-stained prostate tissue were fitted using a time-dependent fluorescence depolarization model. The photoluminescence imaging of Cytate-stained cancerous and normal prostate tissues was accomplished, showing the potential of Cytate as a fluorescence marker for prostate cancer detection.     

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.     

Coregistration of diffuse optical spectroscopy and magnetic resonance imaging in a rat tumor model

We report coregistration of near-infrared diffuse optical spectroscopy (DOS) and magnetic resonance imaging (MRI) for the study of animal model tumors. A combined broadband steady-state and frequency-domain apparatus was used to determine tissue oxyhemoglobin, deoxyhemoglobin, and water concentration locally in tumors. Simultaneous MRI coregistration provided structural (T2-weighted) and contrast-enhanced images of the tumor that were correlated with the optical measurements. By use of Monte Carlo simulations, the optically sampled volume was superimposed on the MR images, showing precisely which tissue structure was probed optically. DOS and MRI coregistration measurements were performed on seven rats over 20 days and were separated into three tumor tissue classifications: viable, edematous, and necrotic. A ratio of water concentration to total hemoglobin concentration, as measured optically, was performed for each tissue type and showed values for edematous tissue to be greater than viable tissue (1.2 +/- 0.49 M/microM versus 0.48 +/- 0.15 M/microM). Tissue hemoglobin oxygen saturation (StO2) also showed a large variation between tissue types: viable tissue had an optically measured StO2 value of 61 +/- 5%, whereas StO2 determined for necrotic tissue was 43 +/- 6%.