K. Thomas Moesta

Assessment of the size, position, and optical properties of breast tumors in vivo by noninvasive optical methods

We present a method for the noninvasive determination of the size, position, and optical properties (absorption and reduced scattering coefficients) of tumors in the human breast. The tumor is first detected by frequency-domain optical mammography. It is then sized, located, and optically characterized by use of diffusion theory as amodel for the propagation of near-infrared light in breast tissue. Our method assumes that the tumor is a spherical inhomogeneity embedded in an otherwise homogeneous tissue. We report the results obtained on a 55-year-old patient with a papillary cancer in the right breast. We found that the tumor absorbs and scatters near-infrared light more strongly than the surrounding healthytissue. Our method has yielded a tumor diameter of 2.1 ? 0.2cm, which is comparable with the actual size of 1.6 cm, determined after surgery. From the tumor absorption coefficients at two wavelengths (690 and 825 nm), we calculated the total hemoglobin concentration (40 ? 10 muM) and saturation (71 ? 9%) of the tumor. These results can provide the clinical examiner with more detailed information about breast lesions detected by frequency-domain optical mammography, thereby enhancing its potential for specificity.

Time-domain optical mammography: initial clinical results on detection and characterization of breast tumors.

Mammograms of 35 patients suspected of breast cancer were taken along craniocaudal and mediolateral projections with a dual-wavelength scanning laser pulse mammograph measuring time-resolved transmittance. Among 26 tumors known from routine clinical diagnostics, 17 tumors were detected retrospectively in optical mammograms. Effective tumor optical properties derived from a homogeneous model were used to deduce physiological information. All tumors exhibited increased total hemoglobin concentration and decreased or unchanged blood oxygen saturation compared with surrounding healthy tissue. Scatter plots based on a pixelwise analysis of individual mammograms were introduced and applied to represent corelations between characteristic quantities derived from measured distributions of times of flight of photons.

Time-domain scanning optical mammography : II. Optical properties and tissue parameters of 87 carcinomas

Within a clinical trial on scanning time-domain optical mammography reported on in a companion publication (part I), craniocaudal and mediolateral projection optical mammograms were recorded from 154 patients, suspected of having breast cancer. Here we report on in vivo optical properties of the subset of 87 histologically validated carcinomas which were visible in optical mammograms recorded at two or three near-infrared wavelengths. Tumour absorption and reduced scattering coefficients were derived from distributions of times of flight of photons recorded at the tumour site employing the model of diffraction of photon density waves by a spherical inhomogeneity, located in an otherwise homogeneous tissue slab. Effective tumour radii, taken from pathology, and tumour location along the compression direction, deduced from off-axis optical scans of the tumour region, were included in the analysis as prior knowledge, if available. On average, tumour absorption coefficients exceeded those of surrounding healthy breast tissue by a factor of about 2.5 (670 nm), whereas tumour reduced scattering coefficients were larger by about 20% (670 nm). From absorption coefficients at 670 nm and 785 nm total haemoglobin concentration and blood oxygen saturation were deduced for tumours and surrounding healthy breast tissue. Apart from a few outliers total haemoglobin concentration was observed to be systematically larger in tumours compared to healthy breast tissue. In contrast, blood oxygen saturation was found to be a poor discriminator for tumours and healthy breast tissue; both median values of blood oxygen saturation are the same within their statistical uncertainties. However, the ratio of total haemoglobin concentration over blood oxygen saturation further improves discrimination between tumours and healthy breast tissue. For 29 tumours detected in optical mammograms recorded at three wavelengths (670 nm, 785 nm, 843 nm or 884 nm), scatter power was derived from transport scattering coefficients. Scatter power of tumours tends to be larger than that of surrounding healthy breast tissue, yet the 95% confidence intervals of both medians overlap.

Concentration and oxygen saturation of haemoglobin of 50 breast tumours determined by time-domain optical mammography

Using a dual-wavelength (670 nm, 785 nm) time-domain scanning instrument we have recorded optical mammograms of 93 patients suspected of having breast cancer which was subsequently assessed histologically. Among 65 histologically confirmed carcinomas, 54 were detectable in at least one of two optical mammograms recorded of each tumour-bearing breast in craniocaudal and mediolateral projection. Optical mammograms were based on photon counts in selected time windows of measured distributions of times of flight of photons. Optical properties of 50 carcinomas investigated at both wavelengths were derived by modelling the breast as partially homogeneous infinite slab with an embedded spherical inhomogeneity representing the tumour and by calculating the diffraction of photon density waves. In selected cases, additional information about the location of the tumour along the compression direction was used that was obtained from scans at selected offsets between source and detector optical fibres. A correlation plot of haemoglobin concentration and blood oxygen saturation of tumours and healthy tissue shows good separation between both kinds of tissue. The majority of carcinomas exhibited increased total haemoglobin concentration compared to healthy tissue.

Frequency-domain optical mammography: Edge effect corrections

We have investigated the problem of edge effects in laser-beam transillumination scanning of the human breast. Edge effects arise from tissue thickness variability along the scanned area, and from lateral photon losses through the sides of the breast. Edge effects can be effectively corrected in frequency-domain measurements by employing a two-step procedure: (1) use of the phase information to calculate an effective tissue thickness for each pixel location; (2) application of the knowledge of tissue thickness to calculate an edge-corrected optical image from the ac signal image. The measurements were conducted with a light mammography apparatus (LIMA) designed for feasibility tests in the clinical environment. Operating in the frequency-domain (110 MHz), this instrument performs a transillumination optical scan at two wavelengths (685 and 825 nm). We applied the proposed two-step procedure to data from breast phantoms and from human breasts. The processed images provide higher contrast and detectability in optical mammography with respect to raw data breast images.

Time-domain scanning optical mammography: I. Recording and assessment of mammograms of 154 patients

Using a triple wavelength (670 nm, 785 nm, 843/884 nm) scanning laser-pulse mammograph we recorded craniocaudal and mediolateral projection optical mammograms of 154 patients, suspected of having breast cancer. From distributions of times of flight of photons recorded at typically 1000-2000 scan positions, optical mammograms were derived displaying (inverse) photon counts in selected time windows, absorption and reduced scattering coefficients or total haemoglobin concentration and blood oxygen saturation. Optical mammograms were analysed by comparing them with x-ray and MR mammograms, including results of histopathology, attributing a subjective visibility score to each tumour assessed. Out of 102 histologically confirmed tumours, 72 tumours were detected retrospectively in both optical projection mammograms, in addition 20 cases in one projection only, whereas 10 tumours were not detectable in any projection. Tumour contrast and contrast-to-noise ratios of mammograms of the same breast, but derived from measured DTOFs by various methods were quantitatively compared. On average, inverse photon counts in selected time windows, including total photon counts, provide highest tumour contrast and contrast-to-noise ratios. Based on the results of the present study we developed a multi-wavelength, multi-projection scanning time-domain optical mammograph with improved spectral and spatial (angular) sampling, that allows us to record entire mammograms simultaneously at various offsets between the transmitting fibre and receiving fibre bundle and provides first results for illustration.

Silencing of human ferrochelatase causes abundant protoporphyrin-IX accumulation in colon cancer

Hemes and heme proteins are vital components of essentially every cell of virtually every eukaryote organism. Previously, we demonstrated accumulation of the heme precursor protoporphyrin‐IX (PplX) in gastrointestinal tumor tissues. To elucidate the mechanisms of PplX accumulation by quantitative reverse transcriptase‐polymerase chain reaction (RT‐PCR), we studied expression of the relevant enzymes of the heme synthetic pathway. Here, we describe a significant down‐regulation of ferrochelatase (FECH) mRNA expression in gastric, colonic, and rectal carcinomas. Accordingly, in an in vitro model of several carcinoma cell lines, ferrochelatase down‐regulation and loss of enzymatic activity corresponded with an enhanced PpIX‐dependent fluorescence. Direct detection of PpIX in minute amounts was achieved by a specifically developed pulsed solid‐state laser dual delay fluorimetry setup. Silencing of FECH using small interfering RNA (siRNA) technology led to a maximum 50‐fold increased PplX accumulation, imageable by a specifically adapted two‐photon microscopy unit. Our results show that in malignant tissue a transcriptional down‐regulation of FECH occurs, which causes endogenous PpIX accumulation. Furthermore, accumulation of intracellular PplX because of FECH siRNA silencing provides a small‐molecule‐based approach to molecular imaging and molecular therap.—Kemmner, W., Wan, K., Rüttinger, S., Ebert, B., Macdonald, R., Klamm, U., Moesta, K. T. Silencing of human ferrochelatase causes abundant protoporphyrin‐IX accumulation in colon cancer. FASEB J. 22, 500–509 (2008)

Scanning time-domain optical mammography : Detection and characterization of breast tumors in vivo

Optical mammography is one of several new techniques for breast cancer detection and characterization presently under development for clinical use that provide information other than morphologic, in particular on the biochemical and metabolic state of normal and diseased tissue. In breast tissue, scattering of red to near infrared (NIR) light dominates absorption and NIR light may penetrate several centimeters through the breast. Optical mammography avoids the use of ionizing radiation and offers the power of diffuse optical spectroscopy. However, because of strong light scattering, spatial resolution of optical mammography is generally low. The paper reviews the results of a clinical study on scanning time-domain optical mammography comprising 154 patients carrying a total of 102 carcinomas validated by histology. Ninety two of these tumors were detected in optical mammograms retrospectively and for 87 of the detected tumors optical properties and tissue parameters were derived. In addition developments on instrumentation and data analysis are covered and possible improvements of optical mammography are briefly discussed.

Quantification of optical properties of a breast tumor using random walk theory

For the first time we use a random walk methodology based on time-dependent contrast functions to quantify the optical properties of breast tumors (invasive ductal carcinoma) of two patients. Previously this theoretical approach was successfully applied for analysis of embedded objects in several phantoms. Data analysis was performed on distributions of times of flight for photons transmitted through the breast which were recorded in vivo using a time-domain scanning mammograph at 670 and 785 nm. The size of the tumors, their optical properties, and those of the surrounding tissue were reconstructed at both wavelengths. The tumors showed increased absorption and scattering. From the absorption coefficients at both wavelengths blood oxygen saturation was estimated for the tumors and the surrounding tissue.

Reconstruction of optical properties of phantom and breast lesion in vivo from paraxial scanning data

We report on the reconstruction of absorption and reduced scattering coefficients of breast tissue in vivo of a patient with mastopathic disease. Distributions of times of flight of photons through the compressed breast were recorded by paraxial scanning. From data measured at four different source-detector offsets optical properties were reconstructed within the linear Rytov approximation by a fast inverse Fourier space method. Low-pass filtering in Fourier space was employed to remove excessive noise from high spatial frequency components and to reduce the computational efforts by a factor of 3, typically. The mammograms displaying reconstructed absorption and reduced scattering coefficients were compared with projection mammograms either obtained by time-window analysis of experimental data or based on average absorption and reduced scattering coefficients which were derived from measured temporal point spread functions within a simple homogeneous model. All inhomogeneities which were visible in the projection mammograms and which could be associated with specific breast tissue compartments could be correlated with inhomogeneities in the reconstructed absorption coefficient. In particular, the mastopathic disease was detected in the reconstructed absorption mammogram. In order to assess reliability of optical properties reconstructed from data obtained by paraxial scanning, corresponding phantom experiments and reconstructions of phantom optical properties were carried out. Because of the limited angular range sampled by the in vivo and phantom measurements, considerable blurring of the absorption coefficient occurs along the compression direction, compromising longitudinal resolution.