An experimental setup for multicolor frequencydomain diffuse optical tomography (FD DOT) was created to visualize neoplasia of breast tissue and to estimate its size. The breast is gently pressed between two glass plates and scanned in the transilluminative configuration by a single source and detector pair. Illumination at three wavelengths (684 nm, 794 nm, and 850 nm) which correspond to different parts of the absorption spectrum in a therapeutic transparency window provides information about concentration of the main absorbers (oxygenated hemoglobin, deoxygenated hemoglobin, and fat/water). Source amplitude modulation at 140 MHz increases spatial resolution and provides separate reconstruction of scattering and absorption coefficients. Moreover, it gives information about breast thickness, which is important for reconstruction. The sensitivity of the system enables one to detect the light propagated through tissue having thickness up to 8 cm. Studies on model media and preliminary in vivo experiments with normal breast and breast carcinoma were performed. An increase of scattering coefficient and total hemoglobin concentration is observed in the tumor area. This corroborates validity of the FD DOT method for breast cancer diagnosis.Examination couch equipped with FD DOT device in the Semashko Regional Hospital (Russia)
A fluorescence diffuse tomography (FDT) setup for monitoring tumor growth in small animals has been created. In this setup an animal is scanned in the transilluminative configuration by a single source and detector pair. To remove stray light in the detection system, we used a combination of interferometric and absorption filters. To reduce the scanning time, an experimental animal was scanned using the following algorithm: (1) large-step scanning to obtain a general view of the animal (source and detector move synchronously); (2) selection of the fluorescing region; and (3) small-step scanning of the selected region and different relative shifts between the source and detector to obtain sufficient information for 3D reconstruction. We created a reconstruction algorithm based on the Holder norm to estimate the fluorophore distribution. This algorithm converges to the solution with a minimum number of fluorescing zones. The use of tumor cell lines transfected with fluorescent proteins allowed us to conduct intravital monitoring studies. Cell lines of human melanomas Mel-P, Mel-Ibr, Mel-Kor, and human embryonic kidney HEK293 Phoenix were transfected with DsRed-Express and Turbo-RFP genes. The emission of red fluorescent proteins (RFPs) in the long-wave optical range permits detection of deep-seated tumors. In vivo experiments were conducted immediately after subcutaneous injection of fluorescing cells into small animals.
Quantum dots (QD) are a new class of fluorescence labels with properties and biomedical applications that are not available with traditional organic dyes and fluorescent proteins. The unique optical properties of QDs (high absorption, quantum yield and emission spanning to near IR) make them attractive as in vivo markers for deep-tissue imaging. Different types of optical tomography allow imaging of fluorescent-labeled tumors located deep in human or animal tissue. In this work, preliminary results of the fluorescent diffuse tomography (FDT) method in application to CdTe/CdSe-core/shell fluorescing nanocrystals are reported. In our experiments we utilized low-frequency amplitude modulation (1 kHz) of the second harmonic of Nd:YAG laser (532 nm). The transilluminative configuration was used in the setup. Results of post mortem experiments with capsules containing QDs placed into esophagus of small animals to simulate labeled tumor are shown. An algorithm of processing fluorescent image based on calculating zero of maximum curvature has been applied to detect fluorescent inclusions boundaries on the image. This work demonstrates potential capabilities of the FDT method for imaging of deep fluorescent tumors in human tissue or animal models of human cancer.Intensity, a.u.
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