In vivo fluorescence lifetime tomography

Three-dimensional diffuse optical imaging of hand joints: System description and phantom studies

Optical imaging of fluorescent probes is an essential tool for investigation of molecular events in small animals for drug developments. In order to get localization and quantification information of fluorescent labels, CEA-LETI has developed efficient approaches in classical reflectance imaging as well as in diffuse optical tomographic imaging with continuous and temporal signals. This paper presents an overview of the different approaches investigated and their performances. High quality fluorescence reflectance imaging is obtained thanks to the development of an original "multiple wavelengths" system. The uniformity of the excitation light surface area is better than 15%. Combined with the use of adapted fluorescent probes, this system enables an accurate detection of pathological tissues, such as nodules, beneath the animal's observed area. Performances for the detection of ovarian nodules on a nude mouse are shown. In order to investigate deeper inside animals and get 3D localization, diffuse optical tomography systems are being developed for both slab and cylindrical geometries. For these two geometries, our reconstruction algorithms are based on analytical expression of light diffusion. Thanks to an accurate introduction of light/matter interaction process in the algorithms, high quality reconstructions of tumors in mice have been obtained. Reconstruction of lung tumors on mice are presented. By the use of temporal diffuse optical imaging, localization and quantification performances can be improved at the price of a more sophisticated acquisition system and more elaborate information processing methods. Such a system based on a pulsed laser diode and a time correlated single photon counting system has been set up. Performances of this system for localization and quantification of fluorescent probes are presented.

This article demonstrates a spectroscopic diffuse optical tomography (DOT) system for the noninvasive continuous imaging of tissue. The system was built on an embedded system architecture along with an algorithm that performs entire system control, data acquisition, noise-free lock-in detection, and wireless data transmission to a host computer, equipped with a graphics processing unit (GPU). The GPU system executes a fast 3-D DOT spectroscopic image reconstruction algorithm. The algorithm computes Jacobians and solves four different depth-sensitive inverse problems of image reconstruction. The host computer sends commands to the DOT server system and collects data, and simultaneously performs high-speed 3-D spectroscopic image reconstruction to display images in real-time. We designed a configurable optical fiber patch to collect depth-resolved diffused light exiting the imaging media by placing the patch on the surface. The simulation and experimental results from two setups showed the spectroscopic imaging capability of the system to map the concentration of oxyhemoglobin, deoxyhemoglobin, and water, and to characterize the tumor located deep inside the tissue. The proposed system uses low-cost LEDs, photodiode detectors, embedded system, and algorithms to perform high-speed 3-D functional imaging and can be further miniaturized. Our promising results pave the way for the development of a low-cost hand-held spectroscopic DOT system for the real-time onsite functional imaging of biological tissue.

Advances in epilepsy studies have shown that specific changes in hemodynamics precede and accompany seizure onset and propagation. However, it has been challenging to noninvasively detect these changes in real time and in humans, due to the lack of fast functional neuroimaging tools. In this study, we present a functional diffuse optical tomography (DOT) method with the guidance of an anatomical human head atlas for 3-dimensionally mapping the brain in real time. Central to our DOT system is a human head interface coupled with a technique that can incorporate topological information of the brain surface into the DOT image reconstruction. The performance of the DOT system was tested by imaging motor tasks-involved brain activities on N = 6 subjects (3 epilepsy patients and 3 healthy controls). We observed diffuse areas of activations from the reconstructed [HbT] images of patients, relative to more focal activations for healthy subjects. Moreover, significant pretask hemodynamic activations were also seen in the motor cortex of patients, which indicated abnormal activities persistent in the brain of an epilepsy patient. This work demonstrates that fast functional DOT is a valuable tool for noninvasive 3-dimensional mapping of brain hemodynamics.

Near-infrared (NIR) fluorescence lifetime imaging (FLI) provides a unique contrast mechanism to monitor biological parameters and molecular events in vivo. Single-photon avalanche diode (SPAD) cameras have been recently demonstrated in FLI microscopy (FLIM) applications, but their suitability for in vivo macroscopic FLI (MFLI) in deep tissues remains to be demonstrated. Herein, we report in vivo NIR MFLI measurement with SwissSPAD2, a large time-gated SPAD camera. We first benchmark its performance in well-controlled in vitro experiments, ranging from monitoring environmental effects on fluorescence lifetime, to quantifying Förster resonant energy transfer (FRET) between dyes. Next, we use it for in vivo studies of target-drug engagement in live and intact tumor xenografts using FRET. Information obtained with SwissSPAD2 was successfully compared to that obtained with a gated intensified charge-coupled device (ICCD) camera, using two different approaches. Our results demonstrate that SPAD cameras offer a powerful technology for in vivo preclinical applications in the NIR window.

Laser diodes are widely used in diffuse optical tomography (DOT) systems but are typically expensive and fragile, while light-emitting diodes (LEDs) are cheaper and are also available in the near-infrared (NIR) range with adequate output power for imaging deeply seated targets. In this study, we introduce a new low-cost DOT system using LEDs of four wavelengths in the NIR spectrum as light sources. The LEDs were modulated at 20 kHz to avoid ambient light. The LEDs were distributed on a hand-held probe and a printed circuit board was mounted at the back of the probe to separately provide switching and driving current to each LED. Ten optical fibers were used to couple the reflected light to 10 parallel photomultiplier tube detectors. A commercial ultrasound system provided simultaneous images of target location and size to guide the image reconstruction. A frequency-domain (FD) laser-diode-based system with ultrasound guidance was also used to compare the results obtained from those of the LED-based system. Results of absorbers embedded in intralipid and inhomogeneous tissue phantoms have demonstrated that the LED-based system provides a comparable quantification accuracy of targets to the FD system and has the potential to image deep targets such as breast lesions.

The performance of the two most promising fluorescence-based temperature sensing techniques, namely the fluorescence intensity ratio (FIR) and fluorescence lifetime (FL) schemes, have been compared. Theoretical calibration graphs for the two methods illustrate the useful monotonic change of the response with temperature variation. Comparison of the responses and the sensitivities of the two schemes show that at very low temperatures the FIR method exhibits a significant variation with temperature, while the response of the FL method becomes constant with its sensitivity approaching zero. With increasing temperature, the FIR and the FL methods (with short relaxation times and shorter intrinsic lifetimes of the upper energy levels) share a similar sensitivity over a wide temperature range. The presence of a long relaxation time or a longer intrinsic lifetime of the upper level in the use of the FL method gives a less satisfactory response. Experimental data obtained for a range of dopant ions in various host materials are found to be consistent with the theoretical expectation, with each material having a specific energy gap difference. The sensitivities of each material are compared graphically which would allow the most appropriate sensor for an intended application to be selected.

We recently applied time domain near infrared diffuse optical spectroscopy (TD-NIRS) to monitor hemodynamics of the cardiac wall (oxy and desoxyhemoglobin concentration, saturation, oedema) on anesthetized swine models. Published results prove that NIRS signal can provide information on myocardial hemodynamic parameters not obtainable with conventional diagnostic clinical tools.1 Nevertheless, the high cost of equipment, acquisition length, sensitivity to ambient light are factors limiting its clinical adoption. This paper introduces a novel approach, based on the use of wavelength and code division multiplexing, applicable to TD-NIRS as well as diffuse optical imaging systems (both topography and tomography); the approach, called WS-CDM (wavelength and space code division mltiplexing), essentially consists of a double stage intensity modulation of multiwavelength CW laser sources using orthogonal codes and their parallel correlation-based decoding after propagation in the tissue; it promises better signal to noise ratio (SNR), higher acquisition speed, robustness to ambient light and lower costs compared to both the conventional systems and the more recent spread spectrum approach based on single modulation with pseudo-random bit sequences (PRBS).2 Parallel acquisition of several wavelengths and from several locations is achievable. TD-NIRS experimental results guided Matlab-based simulations aimed at correlating different coding sequences, lengths, spectrum spreading factor, with the WS-CDM performances on such tissues (achievable SNR, acquisition and reconstruction speed, robustness to channel inequalization, ...). Simulations results and preliminary experimental validation confirm the significant improvements that WS-CDM could bring to diffuse optical imaging (not limited to cardiac functional imaging).

INTRODUCTIONDelirium is associated with mortality and new onset dementia, yet the underlying pathophysiology remains poorly understood. Development of imaging biomarkers has been difficult given the challenging nature of imaging delirious patients. Diffuse optical tomography (DOT) offers a promising approach for investigating delirium given its portability and three‐dimensional capabilities.METHODSTwenty‐five delirious and matched non‐delirious patients (n = 50) were examined using DOT, comparing cerebral oxygenation and functional connectivity in the prefrontal cortex during and after an episode of delirium.RESULTSTotal hemoglobin values were significantly decreased in the delirium group, even after delirium resolution. Functional connectivity between the dorsolateral prefrontal cortex and dorsomedial prefrontal cortex was strengthened post‐resolution compared to during an episode; however, this relationship was still significantly weaker compared to controls.DISCUSSIONThese findings highlight DOT's potential as an imaging biomarker to measure impaired cerebral oxygenation and functional dysconnectivity during and after delirium. Future studies should focus on the role of cerebral oxygenation in delirium pathogenesis and exploring the etiological link between delirium and dementias.HighlightsWe developed a portable diffuse optical tomography (DOT) system for bedside three‐dimensional functional neuroimaging to study delirium in the hospital.We implemented a novel DOT task‐focused seed‐based correlation analysis.DOT revealed decreased cerebral oxygenation and functional connectivity strength in the delirium group, even after resolution of delirium.

Near infrared (NIR) diffused optical tomography (DOT) is emerging as a potential tool of non-invasively diagnosing woman breast cancers, neonatal brain hypoxia, and other human organ diseases. The intensive and worldwide investigations in theory and experiment have revealed the possibility of NIR DOT in providing both anatomical and functional information of biological tissue simultaneously, which is important for distinguishing between healthy and diseased tissues, such as benign and malignant tumors. In this paper, our recent DOT experiments on human lower legs and forearms are presented using our time-resolved measuring system and image reconstruction algorithm based on the modified generalized pulse spectrum technique. It was found that the image quality in the experiments, including both the spatial resolution and the quantitativeness of the targets, was rather poor, and the interior blood vessels undisclosed in the absorption images. To clarify this issue, the influences of target contrast and size on the image reconstruction were investigated with simulated data. We have accordingly obtained the following observations: the quantitativeness of the reconstructed optical properties was prone to be spoiled by the small size ratio and high contrast of the interior targets (such as blood vessels) to the background, and the incompleteness of information embedded in the featured data-types, in addition to the experimental noise, evidently answers for the degradation of the spatial resolution and quantitativeness. It was shown in a further simulative investigation that the image quality could be substantially improved by making full use of the time-resolved data.

In Diffuse Optical Tomography (DOT), data processing and reconstruction stages are crucial to obtain high-quality images. Thus, choosing suitable algorithms for the system is a critical choice. This study aims to determine an appropriate reconstruction algorithm for DOT imaging. There are several reconstruction algorithms used in DOT systems. Some algorithms have been improved for solving specific cases, and some still need to be improved. In this study, we used three algorithms for the reconstruction process: Singular Value Decomposition (SVD), Bi-Conjugated Gradient (Bi-CG), and Transpose Free Quasi Minimal Residual (TFQMR). In testing the algorithms, data of the simulation experiments have been used. The simulation experiments model the tumoral tissue within the breast. All three algorithms were produced correct images while the tumor close to the surface. In the case of the tumor that is not close to the breast surface, the tumor location on the images created by Bi-CG and SVD algorithms was not its actual location. However, the tumor location in the image created by the TFQMR algorithm was close to its actual location. Outcomes of the reconstruction algorithms were evaluated based on correctly defining the location of the tumors by using Mean Percentage Error (MPE), Mean Squared Error (MSE), and Mean Absolute Error (MAE) metrics. We have demonstrated the TFQMR algorithm is a more appropriate reconstruction technique for DOT systems. Thus, we have concluded that TFQMR can have the potential to be used in medical imaging systems.

We present a photoacoustic tomography-guided diffuse optical tomography approach using a hand-held probe for detection and characterization of deeply-seated targets embedded in a turbid medium. Diffuse optical tomography guided by coregistered ultrasound, MRI, and x ray has demonstrated a great clinical potential to overcome lesion location uncertainty and to improve light quantification accuracy. However, due to the different contrast mechanisms, some lesions may not be detectable by a nonoptical modality but yet have high optical contrast. Photoacoustic tomography utilizes a short-pulsed laser beam to diffusively penetrate into tissue. Upon absorption of the light by the target, photoacoustic waves are generated and used to reconstruct, at ultrasound resolution, the optical absorption distribution that reveals optical contrast. However, the robustness of optical property quantification of targets by photoacoustic tomography is complicated because of the wide range of ultrasound transducer sensitivity, the orientation and shape of the targets relative to the ultrasound array, and the uniformity of the laser beam. We show in this paper that the relative optical absorption map provided by photoacoustic tomography can potentially guide the diffuse optical tomography to accurately reconstruct target absorption maps.

We present high spatial density, multi-modal, parallel-plate Diffuse Optical Tomography (DOT) imaging systems for the purpose of breast tumor detection. One hybrid instrument provides time domain (TD) and continuous wave (CW) DOT at 64 source fiber positions. The TD diffuse optical spectroscopy with PMT- detection produces low-resolution images of absolute tissue scattering and absorption while the spatially dense array of CCD-coupled detector fibers (108 detectors) provides higher-resolution CW images of relative tissue optical properties. Reconstruction of the tissue optical properties, along with total hemoglobin concentration and tissue oxygen saturation, is performed using the TOAST software suite. Comparison of the spatially-dense DOT images and MR images allows for a robust validation of DOT against an accepted clinical modality. Additionally, the structural information from co-registered MR images is used as a spatial prior to improve the quality of the functional optical images and provide more accurate quantification of the optical and hemodynamic properties of tumors. We also present an optical-only imaging system that provides frequency domain (FD) DOT at 209 source positions with full CCD detection and incorporates optical fringe projection profilometry to determine the breast boundary. This profilometry serves as a spatial constraint, improving the quality of the DOT reconstructions while retaining the benefits of an optical-only device. We present initial images from both human subjects and phantoms to display the utility of high spatial density data and multi-modal information in DOT reconstruction with the two systems.

In this paper, we present a high performance time-resolved diffuse optical tomography (DOT) system whose working principle and implementation are very different from the conventional methods and features near real-time data acquisition. Dual-wavelength near-infrared light at 784 nm and 808 nm are used as light source. A high bit rate pseudo-random bit sequence (2.488 GB/s) modulates both continuous lights when they go through the external intensity modulator. The diffuse photon density waves responding to such modulated light are detected by array of high sensitivity, high speed silicon avalanche photodetectors (APD). The temporal point spread function can be retrieved very quickly by using hardware auto-/cross-correlation. Our system implementation has achieved system impulse with rise time approaching 0.5 ns which is highly desirable for time-domain DOT application to penetrate depth in few centimeters. The temporal resolution is mainly limited by the photodetectors. System performance is stabilized with accurate temperature control over key components. Experiment results based on tissue-like phantoms verify our design is potential for breast cancer imaging.

Functional neuroimaging is a vital element of neuroscience and cognitive research and, increasingly, is an important clinical tool. Diffuse optical imaging is an emerging, noninvasive technique with unique portability and hemodynamic contrast capabilities for mapping brain function in young subjects and subjects in enriched or clinical environments. We have developed a high-performance, high-density diffuse optical tomography (DOT) system that overcomes previous limitations and enables superior image quality. We show herein the utility of the DOT system by presenting functional hemodynamic maps of the adult human visual cortex. The functional brain images have a high contrast-to-noise ratio, allowing visualization of individual activations and highly repeatable mapping within and across subjects. With the improved spatial resolution and localization, we were able to image functional responses of 1.7 cm in extent and shifts of <1 cm. Cortical maps of angle and eccentricity in the visual field are consistent with retinotopic studies using functional MRI and positron-emission tomography. These results demonstrate that high-density DOT is a practical and powerful tool for mapping function in the human cortex.