Fluorescence Diffuse Optical Tomography Research Articles

A self-normalized, full time-resolved method for fluorescence diffuse optical tomography

A full time-resolved scheme that has been previously applied in diffuse optical tomography is extended to time-domain fluorescence diffuse optical tomography regime, based on a finite-element-finite-time-difference photon diffusion modeling and a Newton-Raphson inversion framework. The merits of using full time-resolved data are twofold: it helps evaluate the intrinsic performance of time-domain mode for improvement of image quality and set up a valuable reference to the assessment of computationally efficient featured-data-based algorithms, and provides a self-normalized implementation to preclude the necessity of the scaling-factor calibration and spectroscopic-feature assessments of the system as well as to overcome the adversity of system instability. We validate the proposed methodology using simulated data, and evaluate its performances of simultaneous recovery of the fluorescent yield and lifetime as well as its superiority to the featured-data one in the fidelity of image reconstruction.

Fluorescence tomography characterization for sub-surface imaging with protoporphyrin IX

Optical imaging of fluorescent objects embedded in a tissue simulating medium was characterized using non-contact based approaches to fluorescence remittance imaging (FRI) and sub-surface fluorescence diffuse optical tomography (FDOT). Using Protoporphyrin IX as a fluorescent agent, experiments were performed on tissue phantoms comprised of typical in-vivo tumor to normal tissue contrast ratios, ranging from 3.5:1 up to 10:1. It was found that tomographic imaging was able to recover interior inclusions with high contrast relative to the background; however, simple planar fluorescence imaging provided a superior contrast to noise ratio. Overall, FRI performed optimally when the object was located on or close to the surface and, perhaps most importantly, FDOT was able to recover specific depth information about the location of embedded regions. The results indicate that an optimal system for localizing embedded fluorescent regions should combine fluorescence reflectance imaging for high sensitivity and sub-surface tomography for depth detection, thereby allowing more accurate localization in all three directions within the tissue.

In vivo mice lung tumor follow-up with fluorescence diffuse optical tomography

We present in vivo experiments conducted with a new fluorescence diffuse optical tomographic (fDOT) system on cancerous mice bearing mammary murine tumors. We first briefly present this new system that has been developed and its associated reconstruction method. Its main specificity is its ability to reconstruct the fluorescence yield even in heterogeneous and highly attenuating body regions such as lungs and to enable mouse inspection without immersion in optical index matching liquid (Intralipid and ink). Some phantom experiments validate the performance of this new system for heterogeneous media inspection. Its use for a mice study is then related. It consists in the follow-up of the lungs at different stages of tumor development after injection of RAFT-(cRGD)4-Alexa700. As expected, the reconstructed fluorescence increases along with the tumor stage. These results validate the use of our system for biological studies of small animals.

基于线性方法的时域荧光扩散光层析实验验证

We present a full three-dimensional, featured-data algorithm for time-domain fluorescence diffuse optical tomography that inverts the Laplace-transformed time-domain coupled diffusion equations and employs a pair of appropriate transform-factors to effectively separate the fluorescent yield and lifetime parameters. By use of a time-correlation single-photon counting system and the normalized Born formulation, we experimentally validate that the proposed scheme can achieve simultaneous reconstruction of the fluorescent yield and lifetime distributions with a reasonable accuracy.

Fluorescence diffuse optical tomography with functional and anatomical a priori information: feasibility study

Fluorescence diffuse optical tomography (FT) is an emerging molecular imaging technique that can spatially resolve both fluorophore concentration and lifetime parameters. In this study, we investigate the performance of a frequency-domain FT system for small inclusions that are embedded in a heterogeneous background. The results demonstrate that functional and structural a priori information is crucial to be able to recover both parameters with high accuracy. The functional a priori information is defined by the absorption and scattering maps at both excitation and emission wavelengths. Similarly, the boundaries of the small inclusion and different regions in the background are utilized as the structural a priori information. Without a priori information, the fluorophore concentration of a 5 mm inclusion in a 40 mm medium is recovered with 50% error, while the lifetime cannot be recovered at all. On the other hand, when both functional and structural information are available, the true lifetime can be recovered and the fluorophore concentration can be estimated only with 5% error. This study shows that a hybrid system that can acquire diffuse optical absorption tomography (DOT), FT and anatomical images in the same setting is essential to be able to recover the fluorophore concentration and lifetime accurately in vivo.

Noncontact fluorescence diffuse optical tomography of heterogeneous media

Fluorescence-enhanced diffuse optical tomography is expected to be useful to the collection of functional information from small animal models. This technique is currently limited by the extent of tissue heterogeneity and management of the shape of the animals. We propose an approach based on the reconstruction of object heterogeneity, which provides an original solution to the two problems. Three evaluation campaigns are described: the first two were performed on phantoms designed to test the reconstructions in highly heterogeneous media and noncontact geometries; the third was conducted on mice with lung tumors to test fluorescence yield reconstruction feasibility in vivo.

Light propagation for time-domain fluorescence diffuse optical tomography by convolution using lifetime function

Time-domain light propagation in biological tissue is studied by solving the forward problem for fluorescence diffuse optical tomography using a convolution of the zero-lifetime emission light and the exponential function for a finite lifetime. We firstly formulate the fundamental equations in a time-domain assuming that the fluorescence lifetime is equal to zero, and then the solution including the lifetime is obtained by convolving the emission light and the lifetime function. The model is a two-dimensional (2-D) 10 mm-radius circle with the optical properties simulating biological tissue for the near infrared light, and contains some inclusions with fluorophores. Temporal and spatial profiles of excitation and emission light are calculated and discussed for several models with different inclusions. The results are physically reasonable and will be used for the inverse problem of fluorescence diffuse optical tomography.

Optimal sparse solution for fluorescent diffuse optical tomography: theory and phantom experimental results

We present a method to accurately localize small fluorescent objects within the tissue using fluorescent diffuse optical tomography (FDOT). The proposed method exploits the localized or sparse nature of the fluorophores in the tissue as a priori information to considerably improve the accuracy of the reconstruction of fluorophore distribution. This is accomplished by minimizing a cost function that includes the L1 norm of the fluorophore distribution vector. Experimental results for a milk-based phantom using a fiber-based cw FDOT system demonstrate the capability of this method in accurately localizing small fluorescent objects deep in the phantom.

Three-dimensional in vivo fluorescence diffuse optical tomography of breast cancer in humans

We present three-dimensional (3D) in vivo images of human breast cancer based on fluorescence diffuse optical tomography (FDOT). To our knowledge, this work represents the first reported 3D fluorescence tomography of human breast cancer in vivo. In our protocol, the fluorophore Indocyanine Green (ICG) is injected intravenously. Fluorescence excitation and detection are accomplished in the soft-compression, parallel-plane, transmission geometry using laser sources at 786 nm and spectrally filtered CCD detection. Phantom and in vivo studies confirm the signals are due to ICG fluorescence, rather than tissue autofluorescence and excitation light leakage. Fluorescence images of breast tumors were in good agreement with those of MRI, and with DOT based on endogenous contrast. Tumorto- normal tissue contrast based on ICG fluorescence was two-to-four-fold higher than contrast based on hemoglobin and scattering parameters. In total the measurements demonstrate that FDOT of breast cancer is feasible and promising.

A Fast Reconstruction Algorithm for Fluorescence Optical Diffusion Tomography Based on Preiteration

Fluorescence optical diffusion tomography in the near-infrared (NIR) bandwidth is considered to be one of the most promising ways for noninvasive molecular-based imaging. Many reconstructive approaches to it utilize iterative methods for data inversion. However, they are time-consuming and they are far from meeting the real-time imaging demands. In this work, a fast preiteration algorithm based on the generalized inverse matrix is proposed. This method needs only one step of matrix-vector multiplication online, by pushing the iteration process to be executed offline. In the preiteration process, the second-order iterative format is employed to exponentially accelerate the convergence. Simulations based on an analytical diffusion model show that the distribution of fluorescent yield can be well estimated by this algorithm and the reconstructed speed is remarkably increased.

Image-guided diffuse optical fluorescence tomography implemented with Laplacian-type regularization

A promising method to incorporate tissue structural information into the reconstruction of diffusion-based fluorescence imaging is introduced. The method regularizes the inversion problem with a Laplacian-type matrix, which inherently smoothes pre-defined tissue, but allows discontinuities between adjacent regions. The technique is most appropriately used when fluorescence tomography is combined with structural imaging systems. Phantom and simulation studies were used to illustrate significant improvements in quantitative imaging and linearity of response with the new algorithm. Images of an inclusion containing the fluorophore Lutetium Texaphyrin (Lutex) embedded in a cylindrical phantom are more accurate than in situations where no structural information is available, and edge artifacts which are normally prevalent were almost entirely suppressed. Most importantly, spatial priors provided a higher degree of sensitivity and accuracy to fluorophore concentration, though both techniques suffer from image bias caused by excitation signal leakage. The use of spatial priors becomes essential for accurate recovery of fluorophore distributions in complex tissue volumes. Simulation studies revealed an inability of the "no-priors" imaging algorithm to recover Lutex fluorescence yield in domains derived from T1 weighted images of a human breast. The same domains were reconstructed accurately to within 75% of the true values using prior knowledge of the internal tissue structure. This algorithmic approach will be implemented in an MR-coupled fluorescence spectroscopic tomography system, using the MR images for the structural template and the fluorescence data for region quantification.

Whole body small animal examination with a diffuse optical tomography instrument

We developed a fluorescent diffuse optical tomography instrument in our laboratory to study in vivo tumour growth without any stress or damage for the animal. By taking into account optical heterogeneities, the reconstruction algorithm allows whole body small animal examination. Experimental results obtained with this system on phantoms and on mice deep lung tumours examination are presented in this paper.

A linear, featured-data scheme for image reconstruction in time-domain fluorescence molecular tomography

Fluorescence diffuse optical tomography (DOT) has attracted many attentions from the community of biomedical imaging, since it provides effective enhancement in imaging contrast. This modality is now rapidly evolving as a potential means of monitoring molecular events in small living organisms with help of molecule-specific contrast agents, referred to as fluorescence molecular tomography (FMT). FMT could greatly promote pathogenesis research, drug development, and therapeutic intervention. Although FMT in steady-state and frequency-domain modes have been heavily investigated, the extension to time-domain scheme is imminent for its several unique advantages over the others. By extending the previously developed generalized pulse spectrum technique for time-domain DOT, we propose a linear, featured-data image reconstruction algorithm for time-domain FMT that can simultaneously reconstruct both fluorescent yield and lifetime images of multiple fluorophores, and validate the methodology with simulated data.

Separately reconstructing the structural and functional parameters of a fluorescent inclusion embedded in a turbid medium

We report a novel imaging technique for fluorescence diffuse optical tomography (FDOT). Unlike conventional FDOT, this technique separates the imaging procedure into two steps to respectively reconstruct the structural information (such as the center position and the radius), and the functional information (such as the fluorophore concentration and/or lifetime) of a fluorescing target embedded in a turbid medium. The structural parameters of the target were estimated from the amplitude ratio and phase difference of fluorescence signals received at different detectors, because the amplitude ratio and phase difference were found independent of, or weakly related to, the functional parameters. Based on the estimated structural parameters, a dual-zone mesh technique was utilized to reconstruct the fluorophore concentration. Results of simulations and phantom experiments showed that the structural parameters could be accurately recovered, without knowing the functional information, and that the reconstruction accuracy of the functional parameter was greater than 80%.

Estimation of kinetic model parameters in fluorescence optical diffusion tomography

We present a technique for reconstructing the spatially dependent dynamics of a fluorescent contrast agent in turbid media. The dynamic behavior is described by linear and nonlinear parameters of a compartmental model or some other model with a deterministic functional form. The method extends our previous work in fluorescence optical diffusion tomography by parametrically reconstructing the time-dependent fluorescent yield. The reconstruction uses a Bayesian framework and parametric iterative coordinate descent optimization, which is closely related to Gauss-Seidel methods. We demonstrate the method with a simulation study.

Contrast-detail analysis characterizing diffuse optical fluorescence tomography image reconstruction

Contrast-detail analysis is used to evaluate the imaging performance of diffuse optical fluorescence tomography (DOFT), characterizing spatial resolution limits, signal-to-noise limits, and the trade-off between object contrast and size. Reconstructed images of fluorescence yield from simulated noisy data were used to determine the contrast-to-noise ratio (CNR). A threshold of CNR=3 was used to approximate a lowest acceptable noise level in the image, as a surrogate measure for human detection of objects. For objects 0.5 cm inside the edge of a simulated tissue region, the smallest diameter that met this criteria was approximately 1.7 mm, regardless of contrast level, and test field diameter had little impact on this limit. Object depth had substantial impact on object CNR, leading to a limit of 4 mm for objects near the center of a 51-mm test field and 8.5 mm for an 86-mm test field. Similarly, large objects near the edge of both test fields required a minimum contrast of 50% to achieve acceptable image CNR. The minimum contrast for large, centered objects ranged between 50% and 100%. Contrast-detail analysis using human detection of lower contrast limits provides fundamentally important information about the performance of reconstruction algorithms, and can be used to compare imaging performance of different systems.

Time Domain Fluorescent Diffuse Optical Tomography: analytical expressions

Light propagation in tissue is known to be favored in the Near Infrared spectral range. Capitalizing on this fact, new classes of molecular contrast agents are engineered to fluoresce in the Near Infrared. The potential of these new agents is vast as it allows tracking non-invasively and quantitatively specific molecular events in-vivo. However, to monitor the bio-distribution of such compounds in thick tissue proper physical models of light propagation are necessary. To recover 3D concentrations of the compound distribution, it is necessary to perform a model based inverse problem: Diffuse Optical Tomography. In this work, we focus on Fluorescent Diffuse Optical Tomography expressed within the normalized Born approach. More precisely, we investigate the performance of Fluorescent Diffuse Optical Tomography in the case of time resolved measurements. The different moments of the time point spread function were analytically derived to construct the forward model. The derivation was performed from the zero order moment to the second order moment. This new forward model approach was validated with simulations based on relevant configurations. Enhanced performance of Fluorescent Diffuse Optical Tomography was achieved using these new analytical solutions when compared to the current formulations.

Fluorescence optical diffusion tomography using multiple-frequency data.

A method is presented for fluorescence optical diffusion tomography in turbid media using multiple-frequency data. The method uses a frequency-domain diffusion equation model to reconstruct the fluorescent yield and lifetime by means of a Bayesian framework and an efficient, nonlinear optimizer. The method is demonstrated by using simulations and laboratory experiments to show that reconstruction quality can be improved in certain problems through the use of more than one frequency. A broadly applicable mutual information performance metric is also presented and used to investigate the advantages of using multiple modulation frequencies compared with using only one.

Thermoacoustic Molecular Imaging of Small Animals

We have designed, constructed, and tested a thermoacoustic computed tomography (TCT) scanner for imaging optical absorption in small animals in three dimensions. The device utilizes pulsed laser irradiation (680-1064 nm) and a unique, 128-element transducer array. We quantified the isotropic spatial resolution of this scanner to be 0.35 mm. We describe a dual-wavelength subtraction technique for isolating optical dyes with TCT. Phantom experiments demonstrate that we can detect 5 fmol of a near-infrared dye (indocyanine green, ICG) in a 1-microL volume using dual-wavelength subtraction. Initial TCT imaging in phantoms and in two sacrificed mice suggests that three-dimensional, optical absorption patterns in small animals can be detected with an order of magnitude better spatial resolution and an order of magnitude better low-contrast detectability in small animals when compared to fluorescence imaging or diffusion optical tomography.

Thermoacoustic molecular imaging of small animals.

We have designed, constructed, and tested a thermoacoustic computed tomography (TCT) scanner for imaging optical absorption in small animals in three dimensions. The device utilizes pulsed laser irradiation (680-1064 nm) and a unique, 128-element transducer array. We quantified the isotropic spatial resolution of this scanner to be 0.35 mm. We describe a dual-wavelength subtraction technique for isolating optical dyes with TCT. Phantom experiments demonstrate that we can detect 5 fmol of a near-infrared dye (indocyanine green, ICG) in a 1-microL volume using dual-wavelength subtraction. Initial TCT imaging in phantoms and in two sacrificed mice suggests that three-dimensional, optical absorption patterns in small animals can be detected with an order of magnitude better spatial resolution and an order of magnitude better low-contrast detectability in small animals when compared to fluorescence imaging or diffusion optical tomography.