Optical properties of selected native and coagulated human brain tissues in vitro in the visible and near infrared spectral range

Medical laser applications require knowledge about the optical properties of target tissue. In this study, the optical properties of selected native and coagulated human brain structures were determined in vitro in the spectral range between 360 and 1100 nm. The tissues investigated included white brain matter, grey brain matter, cerebellum and brainstem tissues (pons, thalamus). In addition, the optical properties of two human tumours (meningioma, astrocytoma WHO grade II) were determined. Diffuse reflectance, total transmittance and collimated transmittance of the samples were measured using an integrating-sphere technique. From these experimental data, the absorption coefficients, the scattering coefficients and the anisotropy factors of the samples were determined employing an inverse Monte Carlo technique. The tissues investigated differed from each other predominantly in their scattering properties. Thermal coagulation reduced the optical penetration depth substantially. The highest penetration depths for all tissues investigated were found in the wavelength range between 1000 and 1100 nm. A comparison with data from the literature revealed the importance of the employed tissue preparation technique and the impact of the theoretical model used to extract the optical coefficients from the measured quantities.     

Optical properties of normal and diseased human breast tissues in the visible and near infrared

The optical absorption and scattering coefficients have been determined for specimens of normal and diseased human breast tissues over the range of wavelengths from 500 to 1100 nm. Total attenuation coefficients were measured for thin slices of tissue cut on a microtome. The diffuse reflectance and transmittance were measured for 1.0 mm thick samples of these tissues, using standard integrating sphere techniques. Monte Carlo simulations were performed to derive the scattering and absorption coefficients, as well as the mean cosine of the scattering angle. The results indicate that scatter exceeds absorption by at least two orders of magnitude. Absorption is most significant at wavelengths below 600 nm. The scattering coefficients lie in the range 30-90 mm-1 at 500 nm, and fall smoothly with increasing wavelength to between 10 and 50 mm-1 at 1100 nm. The scattering coefficient for adipose tissue differs, in that it is invariant with wavelength over this spectral range. For all tissues examined, the scattered light is highly forward peaked, with the mean cosine of the scattering angle in the range 0.945-0.985. Systematic differences between the optical properties of some tissue types are demonstrated.     

Optical properties of human skin in the near infrared wavelength range of 1000 to 2200 nm

In this paper we present the absorption coefficient mu(a) and the isotropic scattering coefficient mu(s)(') for 22 human skin samples measured using a double integrating sphere apparatus in the wavelength range of 1000-2200 nm. These in vitro results show that values for mua) follow 70% of the absorption coefficient of water and values for mu(s)(') range from 3 to 16 cm(-1). From the measured optical properties, it was found that a 2% Intralipid solution provides a suitable skin tissue phantom.     

Immunochemical properties of various fractions of cancerous and normal human tissues

Summary Complete antigen obtained according to Boiven's method, and polysaccharide fractions of saline extracts of cancerous and normal tissues of the human stomach possess antigenic properties. The presence of a protein component in these fractions was established by the method of chromatography on paper. Existence of cancerous specificity in lipid and polysaccharide fractions and in complete antigen of human tumor tissue was demonstrated by the reaction of anaphylaxis with desensitization.Presented by Active Member AMN SSSR N. N. Zhukov-Verezhnikov     

Wavelet-based characterization of spectral fluctuations in normal, benign, and cancerous human breast tissues

Fluorescence intensity fluctuations in the visible wavelength regime in normal, benign, and cancerous human breast tissue samples are studied through wavelet transform. The analyses have been carried out in unpolarized, parallel and perpendicularly polarized channels, for optimal tissue characterization. It has been observed that polarized fluorescence data, particularly the perpendicular components, differentiate various tissue types quite well. Wavelet transform, because of its ability for multiresolution analysis, provides the ideal tool to separate and characterize fluctuations in the fluorescence spectra at different scales. We quantify these differences and find that the fluctuations in the perpendicular channel of the cancerous tissues are more randomized as compared to their normal counterparts. Furthermore, for cancerous tissues, the same is very well described by the normal distribution, which is not the case for normal and benign samples. It has also been observed that, up to a certain point, fluctuations at larger scales are more sensitive to tissue types. The differences in the average, low-pass wavelet coefficients of normal, cancerous, pericanalicular, and intracanalicular benign tissues are also pointed out.     

Comparison of human skin opto-thermal response to near-infrared and visible laser irradiations: a theoretical investigation

Near-infrared wavelengths are absorbed less by epidermal melanin, and penetrate deeper into human skin dermis and blood than visible wavelengths. Therefore, laser irradiation using near-infrared wavelengths may improve the therapeutic outcome of cutaneous hyper-vascular malformations in moderately to heavily pigmented skin patients and those with large-sized blood vessels or blood vessels extending deeply into the skin. A mathematical model composed of a Monte Carlo algorithm to estimate the distribution of absorbed light, numerical solution of a bio-heat diffusion equation to calculate the transient temperature distribution, and a damage integral based on an empirical Arrhenius relationship to quantify the tissue damage was utilized to investigate the optothermal response of human skin to near-infrared and visible laser irradiations in conjunction with cryogen spray cooling. In addition, the thermal effects of a single continuous laser pulse and micropulse-composed laser pulse profiles were compared. Simulation results indicated that a 940 nm wavelength induces improved therapeutic outcome compared with a 585 and 595 nm wavelengths for the treatment of patients with large-sized blood vessels and moderately to heavily pigmented skin. On the other hand, a 585 nm wavelength shows the best efficacy in treating small-sized blood vessels, as characterized by the largest laser-induced blood vessel damage depth compared with 595 and 940 nm wavelengths. Dermal blood content has a considerable effect on the threshold incident dosage for epidermal damage, while the effect of blood vessel size is minimal. For the same macropulse duration and incident dosage, a micropulse-composed pulse profile results in higher peak temperature at the basal layer of skin epidermis than an ideal single continuous pulse profile.     

Optical attenuation characteristics of breast tissues at visible and near-infrared wavelengths.

Optical experiments are described for measuring the attenuation characteristics of breast tissues at visible and near-infrared wavelengths. Total attenuation coefficients post mortem were measured directly in thin tissue sections. They are usually within the range from 10 to 30 mm-1, are rather higher in fat than in fibroglandular specimens and decrease with increasing wavelength. The scattering phase function is strongly forward-peaked with the mean cosine of scattering in the range from 0.85 to 0.97 and appearing more forward-peaked in fat than in fibroglandular tissue. The reduced scattering coefficient is of the order of 1 mm-1 in all tissues. Absorption coefficients were measured indirectly in optically thick sections. They are typically between 0.1 and 0.5 mm-1 at wavelengths around 580 nm and an order of magnitude lower at 850 nm. At 580 nm and shorter wavelengths the absorption in carcinoma is significantly higher than in adjacent uninvolved tissue. Significant differences were observed in the first-order derivatives of the transmission spectra of carcinoma and surrounding tissues at certain infrared wavelengths. Transmission spectra measured in vivo across the wavelength range from 500 to 860 nm have a similar form to the spectra of excised samples. Linear absorption coefficients are generally of the same order of magnitude as those found in vitro although they are lower at green wavelengths.     

Thermal coagulation-induced changes of the optical properties of normal and adenomatous human colon tissues in vitro in the spectral range 400-1,100 nm

The absorption coefficients, the reduced scattering coefficients and the optical penetration depths for native and coagulated human normal and adenomatous colon tissues in vitro were determined over the range of 400-1,100 nm using a spectrophotometer with an internal integrating sphere system, and the inverse adding-doubling method was applied to calculate the tissue optical properties from diffuse reflectance and total transmittance measurements. The experimental results showed that in the range of 400-1,100 nm there were larger absorption coefficients (P < 0.01) and smaller reduced scattering coefficients (P < 0.01) for adenomatous colon tissues than for normal colon tissues, and there were smaller optical penetration depths for adenomatous colon tissues than for normal colon tissues, especially in the near-infrared wavelength. Thermal coagulation induced significant increase of the absorption coefficients and reduced scattering coefficients for the normal and adenomatous colon tissues, and significantly reduced decrease of the optical penetration depths for the normal and adenomatous colon tissues. The smaller optical penetration depth for coagulated adenomatous colon tissues is a disadvantage for laser-induced thermotherapy (LITT) and photodynamic therapy (PDT). It is necessary to adjust the application parameters of lasers to achieve optimal therapy.     

The refractive index of human hemoglobin in the visible range

Because the refractive index of hemoglobin in the visible range is sensitive to the hemoglobin concentration, optical investigations of hemoglobin are important for medical diagnostics and treatment. Direct measurements of the refractive index are, however, challenging; few such measurements have previously been reported, especially in a wide wavelength range. We directly measured the refractive index of human deoxygenated and oxygenated hemoglobin for nine wavelengths between 400 and 700 nm for the hemoglobin concentrations up to 140 g?l(-1). This paper analyzes the results and suggests a set of model functions to calculate the refractive index depending on the concentration. At all wavelengths, the measured values of the refractive index depended on the concentration linearly. Analyzing the slope of the lines, we determined the specific refraction increments, derived a set of model functions for the refractive index depending on the concentration, and compared our results with those available in the literature. Based on the model functions, we further calculated the refractive index at the physiological concentration within the erythrocytes of 320 g?l(-1). The results can be used to calculate the refractive index in the visible range for arbitrary concentrations provided that the refractive indices depend on the concentration linearly.     

In vitro assessment of Macleaya cordata crude extract bioactivity and anticancer properties in normal and cancerous human lung cells

The purpose of this study is to assess the bioactivity and anticancer properties of Macleaya cordata crude extract in vitro using normal fetal lung fibroblast MRC5 and adenocarcinomic epithelial cell A549 as model systems,. Treatment of extract induced cell detachment, rounding, and irregularity in shape, in both normal and adenocarcinomic human lung cells, in accompanied of significant reduction in cell proliferation. The data indicated that necrosis appeared to be involved in compromising cell growth in both types of lung cells since membrane permeability and cell granularity were elevated. Although apoptosis was evident, the responses were differential in normal and diseased lung cells. Viability of treated MRC5 cells was reduced in a dose-dependent manner, demonstrating that the normal lung cells are sensitive to the extract. Surprisingly, A549 viability was slightly elevated in response to extract exposure at low concentration, implying that cells survived were metabolically active; the viability was reduced accordingly to treatment at higher concentrations. The present findings demonstrate that the crude extract of M. cordata contains agents affecting the functioning of normal and diseased lung cells in vitro. The observed cytotoxic effects against adenocarcinomic lung cells validate the potential of using M. cordata as herbal intervention in combined with conventional chemotherapy for lung cancer treatment.     

Non-linear electrical properties of skin in the low frequency range

This paper describes the non-linear electrical properties of the skin during the passage of a sinusoidal current from the standpoint of the constant-current method. The non-linearity occurs in the current dependency of the skin impedance and in Lissajous figures. It exhibits both rapid and slow variations. The concept of a non-linear impedance and its equivalent circuit are introduced for a sinusoidal voltage. With regard to the current dependency of the impedance it can be said that both the starting point and the degree of the dependency vary with frequency and impedance. The nonlinearity is more apparent with a larger current, a lower frequency, and a higher impedance. with an increasing current, the ionic conductance increases and the polarisation admittance decreases. Lissajous figures are formed when the voltage is distorted notably from the sinusoidal wave form. Detailed investigations are undertaken for the elliptic figures, third harmonics, rectification, and breakdown of the skin appearing with a decrease of the frequency and increase of the current. Finally, the special mechanism of the ionic conduction in the keratin layer is indicated as one of the causes for non-linearity.     

Optical properties of platelets and blood plasma and their influence on the optical behavior of whole blood in the visible to near infrared wavelength range

The optical parameters absorption coefficient, scattering coefficient, and the anisotropy factor of platelets (PLTs) suspended in plasma and cell-free blood plasma are determined by measuring the diffuse reflectance, total and diffuse transmission, and subsequently by inverse Monte Carlo simulation. Furthermore, the optical behavior of PLTs and red blood cells suspended in plasma are compared with those suspended in saline solution. Cell-free plasma shows a higher scattering coefficient and anisotropy factor than expected for Rayleigh scattering by plasma proteins. The scattering coefficient of PLTs increases linearly with the PLT concentration. The existence of physiological concentrations of leukocytes has no measurable influence on the absorption and scattering properties of whole blood. In summary, red blood cells predominate over the other blood components by two to three orders of magnitude with regard to absorption and effective scattering. However, substituting saline solution for plasma leads to a significant increase in the effective scattering coefficient and therefore should be taken into consideration.     

Visualization of light propagation in visible Chinese human head for functional near-infrared spectroscopy

Using the visible Chinese human data set, which faithfully represents human anatomy, we visualize the light propagation in the head in detail based on Monte Carlo simulation. The simulation is verified to agree with published experimental results in terms of a differential path-length factor. The spatial sensitivity profile turns out to seem like a fat tropical fish with strong distortion along the folding cerebral surface. The sensitive brain region covers the gray matter and extends to the superficial white matter, leading to a large penetration depth (>3 cm). Finally, the optimal source-detector separation is suggested to be narrowed down to 3-3.5 cm, while the sensitivity of the detected signal to brain activation reaches the peak of 8%. These results indicate that the cerebral cortex folding geometry actually has substantial effects on light propagation, which should be necessarily considered for applications of functional near-infrared spectroscopy.     

Influence of shear rate on the optical properties of human blood in the spectral range 250 to 1100 nm

The intrinsic optical parameters-absorption coefficient mua, scattering coefficient mus, anisotropy factor g, and effective scattering coefficient mus'--are determined for human red blood cells of hematocrit 42.1% dependent on the shear rate in the wavelength range 250 to 1100 nm. Integrating sphere measurements of light transmittance and reflectance in combination with inverse Monte-Carlo simulation are carried out for different wall shear rates between 0 and 1000 s(-1). Randomly oriented cells show maximal mua, mus, and mus' values. Cell alignment and elongation, as well as the Fahraeus effect at increasing shear rates, lead to an asymptotical decrease of these values. The anisotropy factor shows this behavior only below 600 nm, dependent on absorption; above 600 nm, g is almost independent of shear rate. The decrease of mus' is inversely correlated with the hemoglobin absorption. Compared to randomly oriented cells, aggregation reduces all parameters by a different degree, depending on the hemoglobin absorption. It is possible to evaluate the influence of collective scattering phenomena, the absorption within the cell, and the cell shape.     

In vivo expression of the novel CXC chemokine BRAK in normal and cancerous human tissue

Using differential display, we cloned a gene with reduced expression in short-term explants of head and neck squamous cell carcinoma (HNSCC) tumors compared to cultured normal oral epithelial cells. The differentially expressed gene was identical to the recently cloned CXC chemokine BRAK, which is ubiquitously expressed in normal tissue extracts but is absent from many tumor cell lines in vitro. To define the cell populations expressing BRAK in vivo, in situ mRNA hybridization was performed on normal and cancerous tissues from six different histological sites. The predominant normal cell type constitutively expressing BRAK in vivo was squamous epithelium. Expression in tumors was heterogeneous, with the majority of HNSCCs and some cervical squamous cell carcinomas (SCCs) showing loss of BRAK mRNA. Although absent in unstimulated peripheral blood mononuclear cells, high levels of BRAK were consistently found in infiltrating inflammatory cells (with lymphocyte morphology) in nearly all cancers examined. Furthermore, BRAK expression was demonstrated in B cells and monocytes, after stimulation of peripheral blood mononuclear cells with lipopolysaccharide. This study demonstrates for the first time up-regulation of BRAK mRNA by inflammatory cells in the tumor microenvironment and lost expression from certain cancers in vivo. The data suggest that BRAK may have a role in host-tumor interactions.     

Immunohistochemical localization of thrombomodulin in normal human skin and skin tumours

Thrombomodulin (TM) expression has been investigated in sections of normal human skin, in cultured normal human keratinocytes, and in a variety of skin tumours. TM was present in squamous epithelial cells in the spinous layer of normal epidermis and in the outer root sheath of hair follicles, but was absent in the cells of the basal layer. It appeared to be predominantly localized to the cell membrane and the intercellular bridges in these areas. Cultured normal human keratinocytes demonstrated functionally active constitutive TM expression on their cell surface. Immunoperoxidase staining of skin tumours using anti-human TM antibodies demonstrated a typical cell membrane positivity in tumours with squamous or hair follicle differentiation. Basal cell carcinomas showed TM expression only in areas where incomplete squamoid metaplasia occurred. Sweat gland tumours and lesions of the melanogenic system failed to express TM. The localization of TM by immunostaining in various benign and malignant skin tumours typically correlated with their normal skin element of origin. The physiological significance of TM expression in the epidermis is currently undefined.     

Tenascin expression in normal human adult skin and skin appendage tumours

The expression and distribution of tenascin, an extracellular matrix glycoprotein, was investigated immunohistochemically using an anti-human tenascin monoclonal antibody (RCB 1) in formalin-fixed paraffin-embedded tissues obtained from 79 patients with skin appendage tumours, and compared with adjacent normal skin. Tissue specimens were pretreated with actinase and processed by the labelled streptavidin-biotin method. In normal skin, tenascin immunoreactivity was consistently found around the ductal portion of the sweat glands, around the lower part of the hair follicle and hair bulbs, and around or within blood vessels. Immunoreactivity was also observed variably around secretory coils of the sweat glands, and below the epidermis. No immunoreactivity was seen around the sebaceous glands. Tumours originating from sweat glands and hair follicles expressed tenascin around the tumour cells nests, while sebaceous gland tumours were immunonegative. Thus, tenascin expression in skin appendage tumours generally resembled that in corresponding normal tissue.