Jürgen Beuthan

The spatial variation of the refractive index in biological cells

With a phase microscope the phase shift of cells from type L 929 fibroblast and mitochondria from liver cells was measured. Compared to the total phase shift caused by the cell relative to vacuum (approximately 1400 nm) the single phase shift of the mitochondria (approximately 180 nm) is small. Only the nucleus and the membrane of the cell give a visibly different phase shift relative to the mean value of the cell. The Fraunhofer diffraction of the measured phase object is calculated. With a simplified scattering theory, i.e. Rayleigh-Gans Scattering, different phase objects are investigated and their differential cross section is discussed.

Sagittal laser optical tomography for imaging of rheumatoid finger joints.

We present a novel optical tomographic imaging system that was designed to determine two-dimensional spatial distribution of optical properties in a sagittal plane through finger joints. The system incorporates a single laser diode and a single silicon photodetector into a scanning device that records spatially resolved light intensities as they are transmitted through a finger. These data are input to a model-based iterative image reconstruction (MOBIIR) scheme, which uses the equation of radiative transfer (ERT) as a forward model for light propagation through tissue. We have used this system to obtain tomographic images of six proximal interphalangeal finger joints from two patients with rheumatoid arthritis. The optical images were compared to clinical symptoms and ultrasound images.

Near-infrared diffuse optical tomography

Diffuse optical tomography (DOT) is emerging as a viable new biomedical imaging modality. Using near-infrared (NIR) light, this technique probes absorption as well as scattering properties of biological tissues. First commercial instruments are now available that allow users to obtain cross-sectional and volumetric views of various body parts. Currently, the main applications are brain, breast, limb, joint, and fluorescence/bioluminescence imaging. Although the spatial resolution is limited when compared with other imaging modalities, such as magnetic resonance imaging (MRI) or X-ray computerized tomography (CT), DOT provides access to a variety of physiological parameters that otherwise are not accessible, including sub-second imaging of hemodynamics and other fast-changing processes. Furthermore, DOT can be realized in compact, portable instrumentation that allows for bedside monitoring at relatively low cost. In this paper, we present an overview of current state-of-the -art technology, including hardware and image-reconstruction algorithms, and focus on applications in brain and joint imaging. In addition, we present recent results of work on optical tomographic imaging in small animals.

First clinical evaluation of sagittal laser optical tomography for detection of synovitis in arthritic finger joints

Objective: To identify classifiers in images obtained with sagittal laser optical tomography (SLOT) that can be used to distinguish between joints affected and not affected by synovitis. Methods: 78 SLOT images of proximal interphalangeal joints II–IV from 13 patients with rheumatoid arthritis were compared with ultrasound (US) images and clinical examination (CE). SLOT images showing the spatial distribution of scattering and absorption coefficients within the joint cavity were generated. The means and standard errors for seven different classifiers (operator score and six quantitative measurements) were determined from SLOT images using CE and US as diagnostic references. For classifiers showing significant differences between affected and non-affected joints, sensitivities and specificities for various cut off parameters were obtained by receiver operating characteristic (ROC) analysis. Results: For five classifiers used to characterise SLOT images the mean between affected and unaffected joints was statistically significant using US as diagnostic reference, but statistically significant for only one classifier with CE as reference. In general, high absorption and scattering coefficients in and around the joint cavity are indicative of synovitis. ROC analysis showed that the minimal absorption classifier yields the largest area under the curve (0.777; sensitivity and specificity 0.705 each) with US as diagnostic reference. Conclusion: Classifiers in SLOT images have been identified that show statistically significant differences between joints with and without synovitis. It is possible to classify a joint as inflamed with SLOT, without the need for a reference measurement. Furthermore, SLOT based diagnosis of synovitis agrees better with US diagnosis than CE.

In vitro studies and computer simulations to assess the use of a diode laser (850 nm) for laser‐induced thermotherapy (LITT)

This study investigated the feasibility of 850 nm diode laser‐induced thermotherapy (LITT), a new procedure providing local hyperthermia and photocoagulation, a minimally invasive treatment of tumors, and other disorders such as benign prostate hyperplasia.

Multipixel system for gigahertz frequency-domain optical imaging of finger joints

Frequency-domain optical imaging systems have shown great promise for characterizing blood oxygenation, hemodynamics, and other physiological parameters in human and animal tissues. However, most of the frequency domain systems presented so far operate with source modulation frequencies below 150 MHz. At these low frequencies, their ability to provide accurate data for small tissue geometries such as encountered in imaging of finger joints or rodents is limited. Here, we present a new system that can provide data up to 1 GHz using an intensity modulated charged coupled device camera. After data processing, the images show the two-dimensional distribution of amplitude and phase of the light modulation on the finger surface. The system performance was investigated and test measurements on optical tissue phantoms were taken to investigate whether higher frequencies yield better signal-to-noise ratios (SNRs). It could be shown that local changes in optical tissue properties, as they appear in the initial stages of rheumatoid arthritis in a finger joint, are detectable by simple image evaluation, with the range of modulation frequency around 500 MHz proving to yield the highest SNR.

Heat Stress Induced Redistribution of Fluorescent Quantum Dots in Breast Tumor Cells

The probing of living cells in different colors over extended periods of time can be used to see the complicated processes that take place during carcinogenesis or heat stress, for example. Since most therapeutic laser tissue interactions are based on thermal effects a detailed characterization of thermal tissue damages in the cellular and sub-cellular levels is important. In order to study such microdosimetry laser-induced fluorescences of Quantum dots provide a suitable approach. Streptavidin conjugated Qdot™ 605 (Quantum Dot Corp., USA) were used in combination with the concanavalin A-biotin labeling system (Molecular Probes, NL) to observe membrane associated thermal lesions. Fluorescent Qdot conjugates are a promising alternative to organic dyes. The extinction coefficient of Qdot™ 605 streptavidin conjugate is 650,000 M−1 cm−1 at 600 nm. Red fluorescent Qdots™ 605 were selected because autofluorescence of cells in the red spectral range is not relevant. Fluorescence detection was performed with a confocal laser scan microscope LSM410 (Carl Zeiss, Germany). Breast cancer cells were used in the thermal stressing experiments performed at 40°C, 42°C, 45°C, 50°C or 56°C for 30 min, each. In this methodical approach Qdot mediated labeling of heat stressed cells were demonstrated to show alterations of plasma membrane organizations and integrities, respectively.

Frequency-Domain Optical Tomographic Imaging of Arthritic Finger Joints

We are presenting data from the largest clinical trial on optical tomographic imaging of finger joints to date. Overall we evaluated 99 fingers of patients affected by rheumatoid arthritis (RA) and 120 fingers from healthy volunteers. Using frequency-domain imaging techniques we show that sensitivities and specificities of 0.85 and higher can be achieved in detecting RA. This is accomplished by deriving multiple optical parameters from the optical tomographic images and combining them for the statistical analysis. Parameters derived from the scattering coefficient perform slightly better than absorption derived parameters. Furthermore we found that data obtained at 600 MHz leads to better classification results than data obtained at 0 or 300 MHz.

Evolution strategies for optical tomographic characterization of homogeneous media.

The reconstruction problem in diffuse optical tomography can be formulated as an optimization problem, in which an objective function has to be minimized. Current model-based iterative image reconstruction schemes commonly use information about the gradient of the objective function to locate the minimum. These gradient-based search algorithms often find local minima close to an initial guess, or do not converge if the gradient is very small. If the initial guess is too far from the solution, gradient-based schemes prove inefficient for finding the global minimum. In this work we introduce evolution-strategy (ES) algorithms for diffuse optical tomography. These algorithms seek to find the global minimum and are less sensitive to initial guesses and regions with small gradients. We illustrate the fundamental concepts by comparing the performance of gradient-based schemes and ES algorithms in finding optical properties (absorption coefficient microa , scattering coefficient micros , and anisotropy factor g) of a homogenous medium.

Optimal source-modulation frequencies for transport-theory-based optical tomography of small-tissue volumes

In frequency-domain optical tomography (FDOT) the quality of the reconstruction result is affected by the choice of the source-modulation frequency. In general the accuracy of the reconstructed image should improve as the source-modulation frequency increases. However, this is only true for noise-free data. Experimental data is typically corrupted by noise and the accuracy is compromised. Assuming the validity of the widely used shot noise model, one can show that the signal-to-noise ratio (SNR) of the amplitude signal decreases with increasing frequency, whereas the SNR of the phase shift reaches peak values in the range between 400 MHz and 800 MHz. As a consequence, it can be assumed that there exists an optimal frequency for which the reconstruction accuracy would be highest. To determine optimal frequencies for FDOT, we investigate here the frequency dependence of optical tomographic reconstruction results using the frequency-domain equation of radiative transfer. We present numerical and experimental studies with a focus on small tissue volumes, as encountered in small animal and human finger imaging. Best reconstruction results were achieved in the 600-800 MHz frequency range.