Antoine Soubret

Accuracy of fluorescent tomography in the presence of heterogeneities:study of the normalized born ratio

We studied the performance of three-dimensional fluorescence tomography of diffuse media in the presence of heterogeneities. Experimental measurements were acquired using an imaging system consisting of a parallel plate-imaging chamber and a lens coupled charge coupled device camera, which enables conventional planar imaging as well as fluorescence tomography. To simulate increasing levels of background heterogeneity, we employed phantoms made of a fluorescent tube surrounded by several absorbers in different combinations of absorption distribution. We also investigated the effect of low absorbing thin layers (such as membranes). We show that the normalized Born approach accurately retrieves the position and shape of the fluorochrome even at high background heterogeneity. We also demonstrate that the quantification is relatively insensitive to a varying degree of heterogeneity and background optical properties. Findings are further contrasted to images obtained with the standard Born expansion and with a normalized approach that divides the fluorescent field with excitation measurements through a homogeneous medium.

Free-space fluorescence molecular tomography utilizing 360° geometry projections

Fluorescence tomography of diffuse media can yield optimal three-dimensional imaging when multiple projections over 360° geometries are captured, compared with limited projection angle systems such as implementations in the slab geometry. We demonstrate how it is possible to perform noncontact, 360° projection fluorescence tomography of mice using CCD-camera-based detection in free space, i.e., in the absence of matching fluids. This approach achieves high spatial sampling of photons propagating through tissue and yields a superior information content data set compared with fiber-based 360° implementations. Reconstruction feasibility using 36 projections in 10° steps is demonstrated in mice.

Complete-angle projection diffuse optical tomography by use of early photons

We present the first, to our knowledge, experimental images of complex-shaped phantoms embedded in diffuse media by use of optical tomography. Imaging is based on a complete-angle projection tomographic technique that utilizes transmitted early photons. Results are contrasted with measurements obtained at later gates as well as pseudocontinuous-wave data. The scanning system developed employs noncontact illumination and detection technologies that allow for high spatial sampling of transmitted photons. Combining this system with complete-angle illumination is found to be an important strategy toward improved imaging performance, resulting in a better-posed inversion problem. The appropriateness of reconstruction algorithms similar to those employed in x-ray computed tomography are showcased, and suggestions for model improvements are provided.

Planar fluorescence imaging using normalized data

Fluorescence imaging of tissues has gained significant attention in recent years due to the emergence of appropriate reporter technologies that enable noninvasive sensing of molecular function in vivo. Two major approaches have been used so far for fluorescence molecular imaging, i.e., epi-illumination (reflectance) imaging and fluorescence molecular tomography. Transillumination is an alternative approach that has been employed for imaging tissues in the past and could be similarly beneficial for fluorescence molecular imaging. We investigate data normalization schemes in reflectance and transillumination mode and experimentally demonstrate that normalized transillumination offers significant advantages over planar reflectance imaging and over nonnormalized methods. Our observations, based on phantoms and on postmortem and in vivo mouse measurements display image quality improvement, superior depth sensitivity, and improved imaging accuracy over the nonnormalized methods examined. Normalized planar imaging retains implementation simplicity and could be used to improve on standard fluorescence reflectance imaging and as a simplified alternative to the more integrated and accurate tomographic methods.

Fluorescence molecular tomography in the presence of background fluorescence

Fluorescence molecular tomography is an emerging imaging technique that resolves the bio-distribution of engineered fluorescent probes developed for in vivo reporting of specific cellular and sub-cellular targets. The method can detect fluorochromes in picomole amounts or less, imaged through entire animals, but the detection sensitivity and imaging performance drop in the presence of background, non-specific fluorescence. In this study, we carried out a theoretical and an experimental investigation on the effect of background fluorescence on the measured signal and on the tomographic reconstruction. We further examined the performance of three subtraction methods based on physical models of photon propagation, using experimental data on phantoms and small animals. We show that the data pre-processing with subtraction schemes can improve image quality and quantification when non-specific background florescence is present.

Surface Reconstruction for Free-Space 360

Complete projection (360deg) free-space fluorescence tomography of opaque media is poised to enable 3-D imaging through entire small animals in vivo with improved depth resolution compared to 360deg-projection fiber-based systems or limited-view angle systems. This approach can lead to a new generation of fluorescence molecular tomography (FMT) performance since it allows high spatial sampling of photon fields propagating through tissue at any projection, employing nonconstricted animal surfaces. Herein, we employ a volume carving method to capture 3-D surfaces of diffusive objects and register the captured surface in the geometry of an FMT 360deg-projection acquisition system to obtain 3-D fluorescence image reconstructions. Using experimental measurements we evaluate the accuracy of the surface capture procedure by reconstructing the surfaces of phantoms of known dimensions. We then employ this methodology to characterize the animal movement of anaesthetized animals. We find that the effects of animal movement on the FMT reconstructed image were within system resolution limits (~0.07 cm).

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Optical tomography using early photons can improve resolution and reduce the ill-posed nature of the inversion problem. In this work we use 360 degrees projection experimental data to investigate the inversion performance of three commonly used numerical inversion methods: the random algebraic reconstruction technique (rART), singular value decomposition (SVD), and the conjugate-gradient-type method LSQR. Results are contrasted to each other and the effects of different photon propagation models are also investigated. We find that all methods perform adequately given appropriate regularization parameters, and that an experimentally measured photon weight function yields superior results over two approximate weights that have been previously used.

Fluorescence Molecular Tomography and the Effects of Animal Motion

Fluorescence molecular tomography (FMT) is an emerging modality for the in-vivo imaging of fluorescent probes which improves upon existing planar photographic imaging techniques by quantitatively reconstructing fluorochrome distributions in-vivo. We present here results using an FMT system capable of full view imaging for arbitrary surface geometries. Results are presented comparing single and multiple projection configurations, and illustrating the need for properly implemented non-negativity constraints.

Inversion with early photons.

We present the development and performance characteristics of a free-space fluorescence tomography system. The imaging system can capture complete angle projections of photons propagating through tissue in transillumination using a CCD camera. Experimental data on imaging lung cancer are presented. Overall, this imaging approach can offer unprecedented imaging performance in fluorescence molecular tomography of small animals

Analysis of reconstructions in full view fluorescence molecular tomography

Fluorescence tomography is an emerging tool for functional and molecular imaging of tissues. Inversion is however complicated by the high scattering that photons experience when propagating through tissue that reduces resolution. While continuous wave systems offer implementation simplicity, time-resolved systems offer the ability to improve resolution and reduce the ill-posed nature of the inverse problem by using early arriving photons. We present an overview of our current progress in complete-angle illumination early photon tomography and present the associated data processing steps employed for early photon inversions.