Models of light propagation in human tissue applied to cancer diagnostics.

Optical methods such as reflectance and fluorescence spectroscopy are being investigated for their potential to aid cancer detection in a quantitative, minimally invasive manner. Mathematical models of reflectance and fluorescence provide an important link between measured optical data and biomedically-relevant tissue parameters that can be extracted from these data to characterize the presence or absence of disease. The most commonly-used mathematical models in biomedical optics are the diffusion approximation (DA) to the radiative transfer equation, Monte Carlo (MC) computational models of light transport, and semi-empirical models. This paper presents a review of the applications of these models to reflectance and endogenous fluorescence sensing for cancer diagnostics in human tissues. Specific examples are given for cervical, breast, and pancreatic tissues. A comparison of the DA and MC methods in two biologically-relevant regimes of optical parameter space will also be discussed.

Evolution of bubble clouds induced by pulsed cavitational ultrasound therapy - histotripsy.

Mechanical tissue fractionation can be achieved using successive, high-intensity ultrasound pulses in a process termed histotripsy. Histotripsy has many potential clinical applications where noninvasive tissue removal is desired. The primary mechanism for histotripsy is believed to be cavitation. Using fast-gated imaging, this paper studies the evolution of a cavitating bubble cloud induced by a histotripsy pulse (10 and 14 cycles) at peak negative pressures exceeding 21MPa. Bubble clouds are generated inside a gelatin phantom and at a tissue-water interface, representing two situations encountered clinically. In both environments, the imaging results show that the bubble clouds share the same evolutionary trend. The bubble cloud and individual bubbles in the cloud were generated by the first cycle of the pulse, grew with each cycle during the pulse, and continued to grow and collapsed several hundred microseconds after the pulse. For example, the bubbles started under 10 microm, grew to 50 microm during the pulse, and continued to grow 100 microm after the pulse. The results also suggest that the bubble clouds generated in the two environments differ in growth and collapse duration, void fraction, shape, and size. This study furthers our understanding of the dynamics of bubble clouds induced by histotripsy.

Autofluorescence characteristics of immortalized and carcinogen-transformed human bronchial epithelial cells.

Tissue autofluorescence has been explored as a potential method of noninvasive pre-neoplasia (pre-malignancy) detection in the lung. Here, we report the first studies of intrinsic cellular autofluorescence from SV40 immortalized and distinct tobacco-carcinogen-transformed (malignant) human bronchial epithelial cells. These cell lines are useful models for studies seeking to distinguish between normal and pre-neoplastic human bronchial epithelial cells. The cells were characterized via spectrofluorimetry and confocal fluorescence microscopy. Spectrofluorimetry revealed that tryptophan was the dominant fluorophore. No change in tryptophan emission intensity was observed between immortalized and carcinogen-transformed cells. Confocal autofluorescence microscopy was performed using a highly sensitive, spectrometer-coupled instrument capable of limiting emission detection to specific wavelength ranges. These studies revealed two additional endogenous fluorophores, whose excitation and emission characteristics were consistent with nicotinamide adenine dinucleotide (NADH) and flavins. In immortalized human bronchial epithelial cells, the fluorescence of these species was localized to cytoplasmic granules. In contrast, the carcinogen-transformed cells showed an appreciable decrease in the fluorescence intensity of both NADH and flavins and the punctate, spatial localization of the autofluorescence was lost. The observed autofluorescence decrease was potentially the result of changes in the redox state of the fluorophores. The random cytoplasmic fluorescence pattern found in carcinogen-transformed cells may be attributed to changes in the mitochondrial morphology. The implications of these results to pre-neoplasia detection in the lung are discussed.

In vivo NADH fluorescence monitoring as an assay for cellular damage in photodynamic therapy.

In this study the endogenous fluorescence signal attributed to reduced nicotinamide adenine dinucleotide (NADH) has been measured in response to photodynamic therapy (PDT)-induced damage. Measurements on cells in vitro have shown that NADH fluorescence decreased relative to that of controls after treatment with a toxic dose of PDT, as measured within 30 min after treatment. Similarly, assays of cell viability indicated that mitochondrial function was reduced immediately after treatment in proportion to the dose delivered, and the proportion of this dose response did not degrade further over 24 h. Measurements in vivo were used to monitor the fluorescence emission spectrum and the excited state lifetime of NADH in PDT-treated tissue. The NADH signal was defined as the ratio of the integrated fluorescence intensity of the 450 +/- 25 nm emission band relative to the fluorescence intensity integrated over the entire 400-600 nm range of collection. Measurements in murine muscle tissue indicated a 22% reduction in the fluorescence signal immediately after treatment with verteporfin-based PDT, using a dose of 2 mg/kg injected 15 min before a 48 J/cm2 light dose at 690 nm. Control animals without photosensitizer injection had no significant change in the fluorescence signal from laser irradiation at the same doses. This signal was monotonically correlated to the deposited dose used here and could provide a direct dosimetric measure of PDT-induced cellular death in the tissue being treated.

Image analysis for discrimination of cervical neoplasia.

Colposcopy involves visual imaging of the cervix for patients who have exhibited some prior indication of abnormality, and the major goals are to visually inspect for any malignancies and to guide biopsy sampling. Currently colposcopy equipment is being upgraded in many health care centers to incorporate digital image acquisition and archiving. These permanent images can be analyzed for characteristic features and color patterns which may enhance the specificity and objectivity of the routine exam. In this study a series of images from patients with biopsy confirmed cervical intraepithelia neoplasia stage 2/3 are compared with images from patients with biopsy confirmed immature squamous metaplasia, with the goal of determining optimal criteria for automated discrimination between them. All images were separated into their red, green, and blue channels, and comparisons were made between relative intensity, intensity variation, spatial frequencies, fractal dimension, and Euler number. This study indicates that computer-based processing of cervical images can provide some discrimination of the type of tissue features which are important for clinical evaluation, with the Euler number being the most clinically useful feature to discriminate metaplasia from neoplasia. Also there was a strong indication that morphology observed in the blue channel of the image provided more information about epithelial cell changes. Further research in this field can lead to advances in computer-aided diagnosis as well as the potential for online image enhancement in digital colposcopy.

Colonic polyp differentiation using time-resolved autofluorescence spectroscopy.

BACKGROUND: Steady-state autofluorescence spectroscopy has been examined previously as a technique for distinguishing polyp types during colonoscopy. Although time-resolved methods have shown promise for tissue diagnosis in vitro, they have never been applied endoscopically. The aim of this study was to examine the feasibility of performing time-resolved autofluorescence spectroscopy in vivo and to determine the diagnostic accuracy of the technique as applied to colonic dysplasia. METHODS: A time-resolved spectrometer was used to measure the spectrally resolved transient decay of laser-induced autofluorescence emission from colonic tissue in vivo. RESULTS: Seventeen patients with 24 polyps (13 adenomatous, 11 non-adenomatous) were studied. The autofluorescence decay from adenomas was faster than that from non-adenomas. The measured decay time provided a means of distinguishing adenomas from non-adenomas with a sensitivity of 85%, a specificity of 91%, a positive predictive value of 92%, and a negative predictive value of 83%. CONCLUSIONS: Time-resolved autofluorescence spectroscopy is a promising optical diagnostic technique for determining polyp types in vivo.

Initial experience with a real-time video processor for enhancing endoscopic image contrast.

BACKGROUND: An initial experience using a digital video processor to enhance contrast in endoscopic images in real-time is described. METHODS: Endoscopic images were examined with a system using locally adaptive processing to balance variations in illumination and sharpen fine features in real-time. For comparison, indigo carmine dye, a popular exogenous contrast agent, was applied to one patient. RESULTS: In general, contrast-enhanced images appeared to contain more mucosal detail, with reduced glare and features in shadowed areas revealed. When used in conjunction with indigo carmine, the enlarged crypt openings in an aberrant crypt focus were visible, the first such detection in vivo without the use of magnifying endoscopy. CONCLUSIONS: Real-time video contrast enhancement during endoscopic procedures appears to provide excellent mucosal detail and improve contrast in a clinically useful manner.