Inverse Adding-doubling Algorithm Research Articles

Absorption and reduced scattering coefficient estimation in pigmented human skin tissue byexperimental colorimetric fitting.

This study aims to estimate the optical properties, absorption (μ a), and reduced scattering (μ s ') coefficients of ex vivo human skin through the individual typology angle (ITA) by only using the skin color parameters. Human skin samples were grouped according to their ITA value and measured using a colorimeter for validation. An integrating sphere and the inverse adding-doubling algorithm were applied to compute the samples μ a and μ s '. The μ a increases as the ITA decreases. An axis swap was performed to generate the μ a versus the ITA for all wavelengths between 500nm and 800nm with a spectral resolution of 10nm. Linearization was performed and a correlation was found. An equation to fit μ a based solely on the ITA values was estimated. The μ s ' does not change with ITA, but it could be fit with an inverse power law as a function of the wavelength. Both equations have a coefficient of determination (R 2) higher than 0.93, indicating a good agreement with our model. An experimental model to estimate the absorption and reduced scattering coefficients of ex vivo human skin through ITA was found. The model has high agreement with the experimental data, with an R 2 between 0.932 and 0.997, and these findings may be relevant for photobiomodulation and light treatment applications to estimate the effect of the melanin on the therapy.

Optical Properties of Human Skin Phototypes and Their Correlation with Individual Angle Typology

Objective: This study aims to correlate human skin phototypes with complete optical characterization (absorption, scattering, effective attenuation, optical penetration, and albedo coefficients) based on individual typology angle (ITA) values and colorimetric parameters. Methods: A colorimeter was used to group 12, fresh, ex vivo human skin samples according to their phototype; the CIELAB color scale and ITA values were employed. An integrating sphere system and the inverse adding-doubling algorithm were applied during optical characterization, conducted from 500 to 1300 nm. Results: On the basis of ITA values and their classification, the skin samples were separated into six groups: two intermediates, two tan, and two brown. In the visible range, for lower ITA values (darker skins), the absorption and effective attenuation coefficient parameters increased, whereas the albedo and depth penetration parameters decreased. In the infrared region, all the phototypes had similar parameters. The scattering coefficient was similar for all the samples and did not change with ITA values. Conclusions: ITA analysis, a quantitative method, showed that the human skin tissue's optical properties and pigmentation colors were highly correlated.

Rapid extraction of skin physiological parameters from hyperspectral images using machine learning

Noninvasive assessment of skin structure using hyperspectral images has been intensively studied in recent years. Due to the high computational cost of the classical methods, such as the inverse Monte Carlo (IMC), much research has been done with the aim of using machine learning (ML) methods to reduce the time required for estimating parameters. This study aims to evaluate the accuracy and the estimation speed of the ML methods for this purpose and compare them to the traditionally used inverse adding-doubling (IAD) algorithm. We trained three models – an artificial neural network (ANN), a 1D convolutional neural network (CNN), and a random forests (RF) model – to predict seven skin parameters. The models were trained on simulated data computed using the adding-doubling algorithm. To improve predictive performance, we introduced a stacked dynamic weighting (SDW) model combining the predictions of all three individually trained models. SDW model was trained by using only a handful of real-world spectra on top of the ANN, CNN and RF models that were trained using simulated data. Models were evaluated based on the estimated parameters’ mean absolute error (MAE), considering the surface inclination angle and comparing skin spectra with spectra fitted by the IAD algorithm. On simulated data, the lowest MAE was achieved by the RF model (0.0030), while the SDW model achieved the lowest MAE on in vivo measured spectra (0.0113). The shortest time to estimate parameters for a single spectrum was 93.05 μs. Results suggest that ML algorithms can produce accurate estimates of human skin optical parameters in near real-time.

Robustness of diffuse reflectance spectra analysis by inverse adding doubling algorithm.

Analysing diffuse reflectance spectra to extract properties of biological tissue requires modelling of light transport within the tissue, considering its absorption, scattering, and geometrical properties. Due to the layered skin structure, skin tissue models are often divided into multiple layers with their associated optical properties. Typically, in the analysis, some model parameters defining these properties are fixed to values reported in the literature to speed up the fitting process and improve its performance. In the absence of consensus, various studies use different approaches in fixing the model parameters. This study aims to assess the effect of fixing various model parameters in the skin spectra fitting process on the accuracy and robustness of a GPU-accelerated two-layer inverse adding-doubling (IAD) algorithm. Specifically, the performance of the IAD method is determined for noiseless simulated skin spectra, simulated spectra with different levels of noise applied, and in-vivo measured reflectance spectra from hyperspectral images of human hands recorded before, during, and after the arterial occlusion. Our results suggest that fixing multiple parameters to a priori known values generally improves the robustness and accuracy of the IAD algorithm for simulated spectra. However, for in-vivo measured spectra, these values are unknown in advance and fixing optical parameters to incorrect values significantly deteriorates the overall performance. Therefore, we propose a method to improve the fitting performance by pre-estimating model parameters. Our findings could be considered in all future research involving the analysis of diffuse reflectance spectra to extract optical properties of skin tissue.

Accuracy of retrieving optical properties from liquid tissue phantoms using a single integrating sphere.

Using integrating spheres (ISs) in conjunction with the inverse adding-doubling algorithm (IAD) offers a well-established, rigorous protocol for determining optical absorption (μa) and reduced scattering (μs') coefficients of thin, optically homogeneous, turbid media. Here, we report the performance and use of a single IS system for experimentally retrieving optical properties in phantom media whose optical properties were well controlled. The IS system was used to measure the total reflectance and transmittance between 500 and 800 nm in liquid phantoms that were prepared to span a wide range of optical scattering and absorption coefficients. Measurements on phantoms were sequentially made using one of two broadband light sources-a halogen lamp or a supercontinuum laser. We report on the accuracy of IAD-derived optical coefficients using IS measurements made on phantoms-directly or by employing one of two previously reported correction methods. The first (sample-substitution error) correction was experimentally achieved while the second used Monte Carlo-based corrections with IAD. When experimentally calibrated reflectance and transmittance values were directly used as inputs to the IAD, mean absolute errors in recovered optical coefficients were larger than 0.4cm-1 for absorption and more than 6cm-1 for scattering across all phantoms and wavelengths measured. These errors reduced to 0.06-0.17cm-1 and 0.7-2cm-1 for μa and μs', respectively, with the use of corrections. Choice of light sources used, sample geometry (relative to optical coefficients), signal-to-noise of measurements, and the selection of correction methods are discussed.

Polydimethylsiloxane tissue-mimicking phantoms with tunable optical properties

.SignificanceThe polymer, polydimethylsiloxane (PDMS), has been increasingly used to make tissue simulating phantoms due to its excellent processability, durability, flexibility, and limited tunability of optical, mechanical, and thermal properties. We report on a robust technique to fabricate PDMS-based tissue-mimicking phantoms where the broad range of scattering and absorption properties are independently adjustable in the visible- to near-infrared wavelength range from 500 to 850 nm. We also report on an analysis method to concisely quantify the phantoms’ broadband characteristics with four parameters.AimWe report on techniques to manufacture and characterize solid tissue-mimicking phantoms of PDMS polymers. Tunability of the absorption () and reduced scattering coefficient spectra () in the wavelength range of 500 to 850 nm is demonstrated by adjusting the concentrations of light absorbing carbon black powder (CBP) and light scattering titanium dioxide powder (TDP) added into the PDMS base material.ApproachThe and of the phantoms were obtained through measurements with a broadband integrating sphere system and by applying an inverse adding doubling algorithm. Analyses of and of the phantoms, by fitting them to linear and power law functions, respectively, demonstrate that independent control of and is possible by systematically varying the concentrations of CBP and TDP.ResultsOur technique quantifies the phantoms with four simple fitting parameters enabling a concise tabulation of their broadband optical properties as well as comparisons to the optical properties of biological tissues. We demonstrate that, to a limited extent, the scattering properties of our phantoms mimic those of human tissues of various types. A possible way to overcome this limitation is demonstrated with phantoms that incorporate polystyrene microbead scatterers.ConclusionsOur manufacturing and analysis techniques may further promote the application of PDMS-based tissue-mimicking phantoms and may enable robust quality control and quality checks of the phantoms.

Optical investigation of bovine grey and white matters in visible and near-infrared ranges

Abstract Introduction: Due to enormous interests for laser in medicine and biology, optical properties characterization of different tissue have be affecting in development processes. In addition, the optical properties of biological tissues could be influenced by storage methods. Thus, optical properties of bovine white and grey tissues preserved by formalin have been characterized over a wide wavelength spectrum varied between 440 nm and 1000 nm. Materials and Methods: To that end, a single integrating sphere system was assembled for spectroscopic characterization and an inverse adding-doubling algorithm was used to retrieve optical coefficients, i.e. reduced scattering and absorption coefficients. Results: White matter has shown a strong scattering property in comparison to grey matter. On the other hand, the grey matter has absorbed light extensively. In comparison, the reduced scattering profile for both tissue types turned out to be consistent with prior works that characterized optical coefficients in vivo. On the contrary, absorption coefficient behavior has a different feature. Conclusion: Formalin could change the tissue’s optical properties because of the alteration of tissue’s structure and components. The absence of hemoglobin that seeps out due to the use of a formalin could reduce the absorption coefficient over the visible range. Both the water replacement by formalin could reduce the refractive index of a stored tissue and the absence of hemoglobin that scatters light over the presented wavelength range should diminish the reduced scattering coefficients over that wavelength range.

Optical properties in the visible range of two different India ink used as biological phantoms

The optical characteristics of two different brands of India ink, Parker Quink and Pelikan, as an absorber used in preparation a tissue-simulating phantom, have been investigated at visible wavelength range. For this purpose, a single integrating sphere system and a spectrophotometric transmission spectroscopy have been used to estimate the radiometric characteristics. Then, inverse adding-doubling algorithm was implemented to retrieve the optical coefficients of pure ink samples. Parker ink's absorption behavior turned out to incompatible with Pelikan over the present wavelength range. Furthermore, scattering property of both two brands has been exhibited and the albedo was calculated. Parker albedo showed a decremental behavior with wavelength and varied between 0.381 and 0.13. A structural profile of albedo was found for Pelikan, which varied between 0.366 and 0.03. This discrepancy might be explained by the variety of two brands composition besides experimental errors. Furthermore, Parker ink absorption variation turned out to be in correspondence to several tissues' absorption feature over broad wavelength ranges. Finally, Parker ink could be regarded as a better candidate to mimic absorption property for several tissues.

Manufacture and characterization of a 3D-printed integrating sphere

3D printers are more accessible in both academic and research groups, becoming instruments to facilitate the construction of high-quality scientific equipment for laboratories. In the present study, a 3D printed integrating sphere was validated to acquire diffuse reflectance and transmittance on turbid samples in order to compute optical absorption and reduced scattering through the inverse adding doubling algorithm. The 3D printed integrating sphere was validated in three stages: analysis of the integrating sphere coating reflectance using two possible coating materials (barium sulfate powder and a mixture of barium sulfate and paint), the sphere wall reflectance, and using two accurate optical polyurethane phantoms to measure the absorption and reduced scattering coefficients. The results indicate that the system developed with this methodology shows satisfactory reflectance compared to commercial models and is able to acquire experimental diffuse reflectance and transmittance measurements to compute the optical coefficients of turbid samples due to the use of an inverse adding doubling (IAD) algorithm. However, this system is limited to the region from 500 to 1300 nm due to a decrease in the reflectance of the coating.

Analysis of Factors Affecting Optical Performance of GaN-Based Micro-LEDs with Quantum Dots Films

Optical performance in terms of light efficiency, color crosstalk and ambient contrast ratio were analyzed for blue GaN-based micro-light emitting diodes (micro-LEDs) combined with red/green quantum dots (QDs)-polymethyl methacrylate (PMMA) films. The thickness and mass ratio of QDs films are two critical factors in affecting the performance of micro-LEDs. Firstly, the precise optical modeling of QDs-PMMA films is established based on the double integrating sphere (DIS) testing system and inverse adding doubling algorithm (IADA) theory. Red and green QDs-PMMA films are composed of ZnCdSe/ZnS QDs and green ZnCdSeS/ZnS QDs, respectively. The fundamental optical parameters of QDs-PMMA films, including scattering, absorption and anisotropy coefficients, are obtained successfully. Secondly, based on these optical parameters, the Monte Carlo ray tracing method is applied to analyze the effect of a QDs-PMMA film’s thickness and mass ratio on the optical performance of micro-LEDs. Results reveal that the light efficiency first increases and then decreases with the increase of a QDs film’s thickness or mass ratio, owing to the scattering characteristics of QDs. Different from the variation tendencies of light efficiency, the crosstalk between adjacent pixels increases as the QDs-PMMA film’s thickness or mass ratio increases, and the ambient contrast ratio is kept stable when the thickness increases. The mass ratio variation of QDs film can change the optical performance of micro-LEDs more effectively than thickness, which demonstrates that mass ratio is a more important factor affecting the optical performance of micro-LEDs.

Relationship between optical properties and soluble sugar contents of apple flesh during storage

Soluble solids (SS) in fruit are mainly composed of soluble sugars. This research aims to further the understanding of the detection mechanism of soluble solid content (SSC) based on optical technology by exploring the relationship between optical properties and soluble sugar contents. The total reflectance and total transmittance at 400–1050 nm of Fuji apple flesh stored at 25 °C for 50 d and 0 °C for 150 d were collected by an automatic integrating sphere system. The absorption coefficient (μa) and reduced scattering coefficient (μ's) were obtained by iteratively solving the radiative transfer equation using the inverse adding doubling algorithm. The relationship of μa and μ's with the contents of SS, total soluble sugars, fructose, glucose and sucrose were quantitatively analyzed at different wavelengths, and prediction models were established by partial least squares regression (PLSR). The results showed that the changes in μa, μ's, SSC and soluble sugar content presented similar trends during storage at the two test temperatures. As the storage time increased, the decreases in μa and μ's were accompanied by declines in SSC and soluble sugar content. In addition, μa and μ's at 550–1050 nm were both positively correlated with SSC and soluble sugar content, with correlation coefficients (r) of 0.834-0.992 and 0.737-0.981, respectively. Compared with the correlations at 550–780 nm, the correlations at 780–1050 nm between μa and SSC and soluble sugar content were enhanced, while the corresponding correlations with μ's were gradually weakened. In addition, SS was most strongly correlated with sucrose among the three types of soluble sugars. SS and sucrose had closer relationship with μa and μ's than fructose and glucose with μa and μ's. Moreover, their prediction models also performed better than the models for fructose and glucose, with Rp2 values of 0.731-0.804. Thus, the prediction of SSC based on Vis-NIR optical technology may be related to the high correlations between the absorption and scattering properties and the sucrose content.

In Vitro Determination of Concentration-Dependent Optical Properties of Hemoglobin

Abstract The interest of this paper is the determination of the optical properties of oxygenated (saturation above 97 %) hemoglobin in clinical relevant concentrations (ranging from 5 to 15 g/dl), dependent on the layer thickness. Furthermore the generation of a high rate data set for training with machine learning approaches was intended. With a double integrating sphere setup (laser diodes from 780 to 1310 nm) - as a well referenced method - and flow through optical cuvettes ranging from 1 to 3 mm layer thickness, the transmission (𝑀𝑇) and reflection (𝑀𝑅) values of the samples were acquired. From those the layer thickness independent absorption (𝜇𝑎) and reduced scattering coefficients (𝜇𝑠’) were calculated by the means of the Inverse Adding Doubling (IAD) algorithm. For each sample the same coefficients should result correspondingly for all cuvette thicknesses in test. This relationship serves as an internal standard in the evaluation of the collected data sets. In parallel a spectrophotometer in the range from 690 to 1000 nm recorded transmission spectra for all samples as a second reference. First, the IAD algorithm provided optical coefficients (𝜇𝑎, 𝜇𝑠’) in all measurements, with few exceptions at low hemoglobin concentrations. The resulting coefficients match independently of the layer thickness. As a main second result, a high rate data set was generated which serves for further analysis - for example with machine learning approaches.

TISM - a transportable integrating sphere setup for standardized measurements of blood and tissue phantoms

Abstract The development of an optical sensor for non-invasive measurements in humans requires a test setup, where the optical properties of tissue and blood can be adjusted and measured standardized. The goal of this work is to develop a simplified device based on an integrating sphere setup to evaluate the optical properties of tissue and blood phantoms with respect to static as well as flow conditions. Furthermore, the measurement system is intended to be used at different locations such as laboratories and operating theaters. We evaluate the absorption μa and reduced scattering μs' coefficients of specimens, with the developed integrating sphere setup. The measurement is regulated by a microcontroller for averaging and processing the data. The system is housed in a lightproof box and powered by a battery and therefore transportable. Due to this construction, no calibration is necessary between transports of the system. Calculations are executed with the inverse adding doubling algorithm. In order to basically calibrate and evaluate the setup before first transportation, a dilution series with Intralipid and India ink serve for the test. The results were consistent with precedent studies (mean absolute deviation for μs' of 0.75 mm-1) and demonstrate that this method might be able to produce liquids with adjustable optical properties, as required for further research. Furthermore, a first dilution series of heparinized heamoglobin (5 to 15 g/dl) with oxygen saturation of 98 % was measured with this system under flow conditions. We observed a linear increase of μa and μs' with the increment of the haemoglobin concentration. As light sources, laser diodes in the range from 780 to 980 nm were introduced. Static and flow measurements indicated that the system is capable for evaluating optical properties under the selected conditions.

Optical properties of biomimetic probes engineered from erythrocytes

Light-activated theranostic materials offer a potential platform for optical imaging and phototherapeutic applications. We have engineered constructs derived from erythrocytes, which can be doped with the FDA-approved near infrared (NIR) chromophore, indocyanine green (ICG). We refer to these constructs as NIR erythrocyte-mimicking transducers (NETs). Herein, we investigated the effects of changing the NETs mean diameter from micron- (≈4 μm) to nano- (≈90 nm) scale, and the ICG concentration utilized in the fabrication of NETs from 5 to 20 μM on the resulting absorption and scattering characteristics of the NETs. Our approach consisted of integrating sphere-based measurements of light transmittance and reflectance, and subsequent utilization of these measurements in an inverse adding-doubling algorithm to estimate the absorption (μa) and reduced scattering (μs′) coefficients of these NETs. For a given NETs diameter, values of μa increased over the approximate spectral band of 630–860 nm with increasing ICG concentration. Micron-sized NETs produced the highest peak value of μa when using ICG concentrations of 10 and 20 μM, and showed increased values of μs′ as compared to nano-sized NETs. Spectral profiles of μs′ for these NETs showed a trend consistent with Mie scattering behavior for spherical objects. For all NETs investigated, changing the ICG concentration minimally affected the scattering characteristics. A Monte Carlo-based model of light distribution showed that the presence of these NETs enhanced the fluence levels within simulated blood vessels. These results provide important data towards determining the appropriate light dosimetry parameters for an intended light-based biomedical application of NETs.

Double-integrating-sphere system at the National Institute of Standards and Technology in support of measurement standards for the determination of optical properties of tissue-mimicking phantoms.

There is a need for a common reference point that will allow for the comparison of the optical properties of tissue-mimicking phantoms. After a brief review of the methods that have been used to measure the phantoms for a contextual backdrop to our approach, this paper reports on the establishment of a standardized double-integrating-sphere platform to measure absorption and reduced scattering coefficients of tissue-mimicking biomedical phantoms. The platform implements a user-friendly graphical user interface in which variations of experimental configurations and model-based analysis are implemented to compute the coefficients based on a modified inverse adding-doubling algorithm allowing a complete uncertainty evaluation. Repeatability and validation of the measurement results of solid phantoms are demonstrated for three samples of different thicknesses, d = 5.08 mm, 7.09 mm, and 9.92 mm, with an absolute error estimate of 4.0% to 5.0% for the absorption coefficient and 11% to 12% for the reduced scattering coefficient (k = 2). The results are in accordance with those provided by the manufacturer. Measurements with different polarization angles of the incident light are also presented, and the resulting optical properties were determined to be equivalent within the estimated uncertainties.

Optical properties of mouse brain tissue after optical clearing with FocusClear™.

Fluorescence microscopy is commonly used to investigate disease progression in biological tissues. Biological tissues, however, are strongly scattering in the visible wavelengths, limiting the application of fluorescence microscopy to superficial (<200µm) regions. Optical clearing, which involves incubation of the tissue in a chemical bath, reduces the optical scattering in tissue, resulting in increased tissue transparency and optical imaging depth. The goal of this study was to determine the time- and wavelength-resolved dynamics of the optical scattering properties of rodent brain after optical clearing with FocusClear™. Light transmittance and reflectance of 1-mm mouse brain sections were measured using an integrating sphere before and after optical clearing and the inverse adding doubling algorithm used to determine tissue optical scattering. The degree of optical clearing was quantified by calculating the optical clearing potential (OCP), and the effects of differing OCP were demonstrated using the optical histology method, which combines tissue optical clearing with optical imaging to visualize the microvasculature. We observed increased tissue transparency with longer optical clearing time and an analogous increase in OCP. Furthermore, OCP did not vary substantially between 400 and 1000 nm for increasing optical clearing durations, suggesting that optical histology can improve ex vivo visualization of several fluorescent probes.

Effects of temperature-dependent optical properties on the fluence rate and temperature of biological tissue during low-level laser therapy

The effects of temperature-dependent optical properties on the change of fluence rate and temperature distribution within biological tissues during low-level laser therapy (LLLT) were investigated by experimental and numerical methods. The fluence rate and temperature within a porcine skin were measured in vitro using an optical fiber sensor and a thermocouple, respectively, while irradiating the sample with a continuous wave laser (IPG Laser GmbH, Burbach, Germany, 1,064nm, 3.14W/cm(2)). The absorption and reduced scattering coefficients of porcine skin were estimated using an inverse adding-doubling algorithm from the total reflectance and transmittance measured with a double-integrating sphere. It was shown that the reduced scattering coefficient of porcine skin decreased significantly as the skin temperature increased within the range of 26-40°C. To incorporate the temperature dependency of tissue optical properties in the simulation, a mathematical model that adopted coupled equations for fluence rate and bioheat transfer was developed. It was shown that the predicted fluence rate and temperature by the proposed mathematical model agreed closely with the measured values of porcine skin. The calculation of human skin temperature using the developed model revealed that the skin temperature could be significantly underestimated if the temperature dependency of optical properties of human skin were ignored during LLLT simulation.

Optical properties of breast tumor phantoms containing carbon nanotubes and nanohorns.

The degree by which optical properties of tumors are altered following introduction of carbon nanotubes (CNTs) of varying concentration and type is poorly understood, making it difficult to predict the impact of CNT inclusion on the photothermal response to laser therapies. Optical properties were measured of phantoms representative of breast tumor tissue incorporated with multiwalled carbon nanotubes (MWNTs), single-walled carbon nanotubes (SWNTs), and single-walled carbon nanohorns (SWNHs) of varying concentration (0.01-0.1 mg/ml). Tissue phantoms were made from sodium alginate (3 g/ml) incorporated with polystyrene microbeads (3 μm diam and 1 mg/ml) and talc-France powder (40 mg/ml). Absorption (μ(a)) and reduced scattering (μ's) coefficients of phantoms containing CNTs were determined by the inverse adding-doubling algorithm for the wavelength range of 400-1300 nm. Optical properties of phantoms without CNTs were in the range of μ(a) = 1.04-0.06 mm(-1) and μ's' = 0.05-0.07 mm(-1) at a wavelength of 900 nm, which corresponds with published data for human breast tumor tissue. Incorporating MWNTs, SWNTs, and SWNHs in phantoms with a concentration of 0.1 mg/ml increased (μ(a)) by 20- to 30-fold, 5- to 6-fold, and 9- to 14-fold, respectively, for the wavelength range of 800-1100 nm with minimal change in μ's (1.2- to 1.3-fold). Introduction of CNTs into tissue phantoms increased absorption, providing a means to enhance photothermal therapy.

Pulsed-laser ablation of polytetrafluoroethylene (PTFE) at various wavelengths

The ablation rate of sintered polytetrafluoroethylene (PTFE) targets has been investigated as a function of wavelength within the range nm and pulse durations ns by means of various different lasers. In this parameter range, the apparent optical properties of targets and the depth of energy deposition are determined mainly by the scattering of the laser light. Therefore, measurements of the scattered light were performed for samples of various thicknesses by means of a modified UV-VIS spectrometer. For selected wavelengths, the angular distribution of the reflected and transmitted light intensity was determined. From these measurements, the coefficients for linear absorption, linear scattering and the scattering anisotropy were calculated using the inverse adding doubling algorithm. The results were tested by Monte Carlo simulation. With these coefficients the apparent optical penetration depths were estimated. These are about one order of magnitude larger than the ablation depths per pulse, which were determined from the weight loss of the targets. Laser deposition

Use of a blood substitute to determine instantaneous murine right ventricular thickening with optical coherence tomography.

A satisfactory imaging technique to determine regional wall thickening of the murine myocardium is not available. Although cardiovascular imaging with light offers a novel solution, application is problematic because scattering by erythrocytes causes significant optical attenuation. Optical coherence tomography (OCT) is a technique for detailed resolution imaging of highly scattering biological tissues. To reduce the high level of blood scattering, a method was devised whereby murine blood was replaced with a hemoglobin-based blood substitute. The scattering and absorption properties of in vitro preparations of whole blood and dilutions of blood with a blood substitute were determined with a spectrophotometer and an inverse-adding doubling algorithm. OCT imaging of the same dilutions demonstrated a significant reduction in scattering at a hematocrit <5%. A fiber-optic OCT imaging system was used to image the murine right midventricular free wall before and after isovolumic replacement with blood substitute. Strong light attenuation prevented full thickness imaging before replacement, whereas visualization of the full ventricular thickness was possible after replacement. Baseline and imaging hematocrits were 52.4+/-3.8% and 3.7+/-1.2%, respectively. End-systolic and end-diastolic thickness values were 0.458+/-0.051 mm and 0.352+/-0.047 mm. Percent thickening fraction was 30.8+/- 7.5%. Optical imaging of the intact beating murine right ventricle was substantially improved by isovolumic blood replacement with a hemoglobin-based blood substitute. Although the current study has been directed toward imaging the murine heart, a blood substitute may be applied to various optical diagnostic and therapeutic techniques under investigation in cardiovascular medicine.