Liquid Tissue Phantoms Research Articles

Engineering vascularized skin-mimetic phantom for non-invasive Raman spectroscopy

Recent advances in Raman spectroscopy have shown great potential for non-invasive analyte sensing, but the lack of a standardized optical phantom for these measurements has hindered further progress. While many research groups have developed optical phantoms that mimic bulk optical absorption and scattering, these materials typically have strong Raman scattering, making it difficult to distinguish metabolite signals. As a result, solid tissue phantoms for spectroscopy have been limited to highly scattering tissues such as bones and calcifications, and metabolite sensing has been primarily performed using liquid tissue phantoms. To address this issue, we have developed a layered skin-mimetic phantom that can support metabolite sensing through Raman spectroscopy. Our approach incorporates millifluidic vasculature that mimics blood vessels to allow for diffusion akin to metabolite diffusion in the skin. Furthermore, our skin phantoms are mechanically mimetic, providing an ideal model for development of minimally invasive optical techniques. By providing a standardized platform for measuring metabolites, our approach has the potential to facilitate critical developments in spectroscopic techniques and improve our understanding of metabolite dynamics in vivo.

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.

Ultracompact fluorescence smartphone attachment using built-in optics for protoporphyrin-IX quantification in skin.

Smartphone-based fluorescence imaging systems have the potential to provide convenient quantitative image guidance at the point of care. However, common approaches have required the addition of complex optical attachments, which reduce translation potential. In this study, a simple clip-on attachment appropriate for fluorescence imaging of protoporphyrin-IX (PpIX) in skin was designed using the built-in light source and ultrawide camera sensor of a smartphone. Software control for image acquisition and quantitative analysis was developed using the 10-bit video capability of the phone. Optical performance was characterized using PpIX in liquid tissue phantoms and endogenously produced PpIX in mice and human skin. The proposed system achieves a very compact form factor (<30 cm3) and can be readily fabricated using widely available low-cost materials. The limit of detection of PpIX in optical phantoms was <10 nM, with good signal linearity from 10 to 1000 nM (R2 >0.99). Both murine and human skin imaging verified that in vivo PpIX fluorescence was detected within 1 hour of applying aminolevulinic acid (ALA) gel. This ultracompact handheld system for quantification of PpIX in skin is well-suited for dermatology clinical workflows. Due to its simplicity and form factor, the proposed system can be readily adapted for use with other smartphone devices and fluorescence imaging applications. Hardware design and software for the system is made freely available on GitHub (https://github.com/optmed/CompactFluorescenceCam).

Multi-Spectral Clinical Prototype for Fluorophore Detection

Identification of tumour margins during resection of the brain is critical for improving the post-operative outcomes. Current methods of tumour identification use 5-ALA, an exogenous precursor, metabolized to fluorescent PpIX in tumour tissue. Although visible under fluorescent microscope, PpIX is easily photo-bleached and tumour tagging is subjective, resulting in tumour under-resection and accelerated recurrence. To address this issue, photo-bleaching resistant and quantitative method is required. This study describes the characterization of a pulsed, multi-wavelengths system designed to measure diffuse reflectance and auto-fluorescence under strong ambient illumination conditions. The performance was tested on n = 400 liquid tissue phantoms containing a wide concentration range of absorber, scatterer and two fluorophores as well as on ex-vivo samples of gray and white matter. The background subtraction technique was shown to be efficient for a range of ambient illumination intensities. A linear relationship was observed between system response and predicted fluorophore concentrations as well as 97.8% accuracy of tissue classification by 5-fold cross-correlation, linear SVM.

Multiband antenna design based on Gosper fractal for implantable biomedical devices

AbstractIn this paper, a novel multiband implantable planar inverted-F antenna (PIFA) antenna based on Gosper curve fractal geometry is designed for biomedical telemetry applications. The antenna has covered four dedicated frequency bands; medical implant communication system band (MICS 402–405 MHz), industrial, scientific, and medical bands (ISM 902–928 MHz and 2.4–2.5 GHz), and wireless medical telemetry services (WMTS 1395–1400 and 1429–1432 MHz). The proposed antenna is designed on Rogers RO 3010 substrate of thickness 25 mil and volume equal to 153.67 mm3. The reflection coefficient and the radiation pattern are measured inside muscle-mimicking liquid tissue phantom. The antenna has achieved the impedance bandwidth of 126, 406, 168, and 175 MHz at MICS (403 MHz), ISM (915 MHz), WMTS (1400 MHz), and ISM (2.45 GHz) with maximum gain value −33.6, −21.04, −15.48, and −10.25 dBi, respectively. The data link range between the implantable antenna and off body antenna has been performed with −16 dBm input power. Additionally, the obtained specific absorption rate with the input of 25 μW power has also been obtained within the safety limit and hence ensures the reliability of the proposed antenna.

Fluorescence and Treatment Light Monitoring for Interstitial Photodynamic Therapy.

In interstitial photodynamic therapy, light is distributed to the tumor via light diffusers. The light dose and the related phototoxic effect achieved throughout the target volume critically depend on absorption, scattering and diffuser positioning. Using liquid tissue phantoms, we investigated the dependencies of treatment light transmission and protoporphyrin IX (PpIX) fluorescence on these parameters. This enabled monitoring hemoglobin oxygenation and methemoglobin formation during irradiation (635nm, 200 mWcm-1 diffuser length). Starting with two parallel cylindrical diffusers at 10mm radial separation, the light transmitted between the fibers was largely determined by the minimal distance between the diffusers, but rather insensitive to an additional axial displacement or tilting of one fiber with respect to the other. For fixed distance between the diffusor centers, however, tilting up to direct contact resulted in a 10-fold signal increase. For hemoglobin within erythrocytes, irradiation leads to photobleaching of PpIX without marked change in hemoglobin oxygenation until hemolysis occurs. Afterward, hemoglobin is rapidly deoxygenized and methemoglobin is formed, leading to a dramatic increase in absorption. For lysed blood, these effects start immediately. A comparison of intraoperative monitoring of the signals with the experimental results might help prevent insufficient treatment by reconsidering treatment planning or prolonging irradiation.

Methods of extraction of optical properties from diffuse reflectance measurements of ex-vivo human colon tissue using thin film silicon photodetector arrays.

Spatially resolved diffuse reflectance spectroscopy (SRDRS) is a promising technique for characterization of colon tissue. Herein, two methods for extracting the reduced scattering and absorption coefficients ( and ) from SRDRS data using lookup tables of simulated diffuse reflectance are reported. Experimental measurements of liquid tissue phantoms performed with a custom multi-pixel silicon SRDRS sensor spanning the 450 - 750 nm wavelength range were used to evaluate the extraction methods, demonstrating that the combined use of spatial and spectral data reduces extraction error compared to use of spectral data alone. Additionally, SRDRS measurements of normal and tumor ex-vivo human colon tissue are presented along with and extracted from these measurements.

Measuring light transport properties using speckle patterns as structured illumination

The measurement of light absorption and scattering properties of biological materials has several diagnostic and therapeutic applications. We can measure these properties for skin without contact using structured illumination and imaging. However, building simple handheld devices remains challenging due to motion artefacts and moving targets. To overcome this limitation, we project random speckle patterns instead of discrete spatial frequencies on the target. Since random patterns are spatially broadband, they capture more information per image, enabling frame-by-frame analysis. In this paper, we describe the statistics of objective speckles and demonstrate how the optical system is designed for spatially bandlimited illumination. Next, we use a diverse set of liquid tissue phantom to validate the method. We successfully demonstrate that a calibrated instrument can independently predict the two primary light transport properties of a homogeneous turbid system. This work is a starting point for analysing skin and other heterogeneous biological media in the future.

NADH-fluorescence scattering correction for absolute concentration determination in a liquid tissue phantom using a novel multispectral magnetic-resonance-imaging-compatible needle probe

In this report, a quantitative nicotinamide adenine dinucleotide hydrate (NADH) fluorescence measurement algorithm in a liquid tissue phantom using a fiber-optic needle probe is presented. To determine the absolute concentrations of NADH in this phantom, the fluorescence emission spectra at 465 nm were corrected using diffuse reflectance spectroscopy between 600 nm and 940 nm. The patented autoclavable Nitinol needle probe enables the acquisition of multispectral backscattering measurements of ultraviolet, visible, near-infrared and fluorescence spectra. As a phantom, a suspension of calcium carbonate (Calcilit) and water with physiological NADH concentrations between 0 mmol l−1 and 2.0 mmol l−1 were used to mimic human tissue. The light scattering characteristics were adjusted to match the backscattering attributes of human skin by modifying the concentration of Calcilit. To correct the scattering effects caused by the matrices of the samples, an algorithm based on the backscattered remission spectrum was employed to compensate the influence of multiscattering on the optical pathway through the dispersed phase. The monitored backscattered visible light was used to correct the fluorescence spectra and thereby to determine the true NADH concentrations at unknown Calcilit concentrations. Despite the simplicity of the presented algorithm, the root-mean-square error of prediction (RMSEP) was 0.093 mmol l−1.

Optical Characterization of Tissue Phantoms Using a Silicon Integrated fdNIRS System on Chip.

An interface circuit with signal processing and digitizing circuits for a high frequency, large area avalanche photodiode (APD) has been integrated in a 130 nm BiCMOS chip. The system enables the absolute oximetry of tissue using frequency domain Near Infrared Spectroscopy (fdNIRS). The system measures the light absorbed and scattered by the tissue by measuring the reduction in the amplitude of signal and phase shift introduced between the light source and detector which are placed a finite distance away from each other. The received 80 MHz RF signal is downconverted to a low frequency and amplified using a heterodyning scheme. The front-end transimpedance amplifier has a 3-level programmable gain that increases the dynamic range to 60 dB. The phase difference between an identical reference channel and the optical channel is measured with a 0.5° accuracy. The detectable current range is [Formula: see text] and with a 40 A/W reponsivity using the APD, power levels as low as 500 pW can be detected. Measurements of the absorption and reduced scattering coefficients of solid tissue phantoms using this system are compared with those using a commercial instrument with differences within 30%. Measurement of a milk based liquid tissue phantom show an increase in absorption coefficient with addition of black ink. The miniaturized circuit serves as an efficiently scalable system for multi-site detection for applications in neonatal cerebral oximetry and optical mammography.

Characterisation of signal enhancements achieved when utilizing a photon diode in deep Raman spectroscopy of tissue.

We characterise the performance of a beam enhancing element ('photon diode') for use in deep Raman spectroscopy (DRS) of biological tissues. The optical component enhances the number of laser photons coupled into a tissue sample by returning escaping photons back into it at the illumination zone. The method is compatible with transmission Raman spectroscopy, a deep Raman spectroscopy concept, and its implementation leads to considerable enhancement of detected Raman photon rates. In the past, the enhancement concept was demonstrated with a variety of samples (pharmaceutical tablets, tissue, etc) but it was not systematically characterized with biological tissues. In this study, we investigate the enhancing properties of the photon diode in the transmission Raman geometry as a function of: a) the depth and b) the optical properties of tissue samples. Liquid tissue phantoms were employed to facilitate systematic variation of optical properties. These were chosen to mimic optical properties of human tissues, including breast and prostate. The obtained results evidence that a photon diode can enhance Raman signals of tissues by a maximum of × 2.4, although it can also decrease the signals created towards the back of samples that exhibit high scattering or absorption properties.

Optimal estimation reconstruction of the optical properties of a two-layered tissue phantom from time-resolved single-distance measurements.

In this work, we have tested the optimal estimation (OE) algorithm for the reconstruction of the optical properties of a two-layered liquid tissue phantom from time-resolved single-distance measurements. The OE allows a priori information, in particular on the range of variation of fit parameters, to be included. The purpose of the present investigations was to compare the performance of OE with the Levenberg–Marquardt method for a geometry and real experimental conditions typically used to reconstruct the optical properties of biological tissues such as muscle and brain. The absorption coefficient of the layers was varied in a range of values typical for biological tissues. The reconstructions performed demonstrate the substantial improvements achievable with the OE provided a priori information is available. We note the extreme reliability, robustness, and accuracy of the retrieved absorption coefficient of the second layer obtained with the OE that was found for up to six fit parameters, with an error in the retrieved values of less than 10%. A priori information on fit parameters and fixed forward model parameters clearly improves robustness and accuracy of the inversion procedure.

Characterisation of a fibre optic Raman probe within a hypodermic needle.

We demonstrate the first use of a multifibre Raman probe that fits inside the bore of a hypodermic needle. A Raman probe containing multiple collection fibres provides improved signal collection efficiency in biological samples compared with a previous two-fibre design. Furthermore, probe performance (signal-to-noise ratios) compared favourably with the performance achieved in previous Raman microscope experiments able to distinguish between benign lymph nodes, primary malignancies in lymph nodes and secondary malignancies in lymph nodes. The experimental measurements presented here give an indication of the sampling volume of the Raman needle probe in lymphoid tissues. Liquid tissue phantoms were used that contained scattering medium encompassing a range of scattering properties similar to those of a variety of tissue types, including lymph node tissues. To validate the appropriateness of the phantoms, the sampling depth of the probe was also measured in excised lymph node tissue. More than 50% of Raman photons collected were found to originate from between the tip of the needle and a depth of 500μm into the tissue. The needle probe presented here achieves spectral quality comparable to that in numerous studies previously demonstrating Raman disease discrimination. It is expected that this approach could achieve targeted subcutaneous tissue measurements and be viable for use for the in vivo Raman diagnostics of solid organs located within a few centimetres below the skin's surface. Graphical Abstract Schematic of multi-fibre Raman needle probe with disposible tips and proximal optical filtration.

Studying the distribution of deep Raman spectroscopy signals using liquid tissue phantoms with varying optical properties.

In this study we employed large volume liquid tissue phantoms, consisting of a scattering agent (Intralipid), an absorption agent (Indian ink) and a synthesized calcification powder (calcium hydroxyapatite (HAP)) similar to that found in cancerous tissues (e.g. breast and prostate), to simulate human tissues. We studied experimentally the magnitude and origin of Raman signals in a transmission Raman geometry as a function of optical properties of the medium and the location of calcifications within the phantom. The goal was to inform the development of future noninvasive cancer screening applications in vivo. The results provide insight into light propagation and Raman scattering distribution in deep Raman measurements, exploring also the effect of the variation of relative absorbance of laser and Raman photons within the phantoms. Most notably when modeling breast and prostate tissues it follows that maximum signals is obtained from the front and back faces of the tissue with the central region contributing less to the measured spectrum.

High-Resolution Fluorescence Diffuse Optical Tomography Developed with Nonlinear Upconverting Nanoparticles

Fluorescence diffuse optical tomography (FDOT) is an emerging biomedical imaging technique that can be used to localize and quantify deeply situated fluorescent molecules within tissues. However, the potential of this technique is currently limited by its poor spatial resolution. In this work, we demonstrate that the current resolution limit of FDOT can be breached by exploiting the nonlinear power-dependent optical emission property of upconverting nanoparticles doped with rare-earth elements. The rare-earth-doped core-shell nanoparticles, NaYF(4):Yb(3+)/Tm(3+)@NaYF(4) of hexagonal phase, are synthesized through a stoichiometric method, and optical characterization shows that the upconverting emission of the nanoparticles in tissues depends quadratically on the power of excitation. In addition, quantum-yield measurements of the emission from the synthesized nanoparticles are performed over a large range of excitation intensities, for both core and core-shell particles. The measurements show that the quantum yield of the 800 nm emission band of core-shell upconverting nanoparticles is 3.5% under an excitation intensity of 78 W/cm(2). The FDOT reconstruction experiments are carried out in a controlled environment using liquid tissue phantoms. The experiments show that the spatial resolution of the FDOT reconstruction images can be significantly improved by the use of the synthesized upconverting nanoparticles and break the current spatial resolution limits of FDOT images obtained from using conventional linear fluorophores as contrast agents.

Lock-in 증폭기를 채용한 주파수영역 확산 광단층촬영 시스템

In this paper, we developed a frequency-domain diffuse optical tomography(DOT) system for non-invasively imaging in vivo. The system uses near-infrared(NIR) light sources and detectors for which the photon propagation in human tissue is dominated by scattering rather than by absorption. We present the experimental reconstruction images of absorption and scattering coefficients using a liquid tissue phantom, and we obtain the location and shape of an anomaly which has different optical properties than the phantom.

Estimation of regional hemoglobin concentration in biological tissues using diffuse reflectance spectroscopy with a novel spectral interpretation algorithm

Both in medical research and clinical settings, regional hemoglobin concentrations ([Hb]) in the microcirculation of biological tissues are highly sought. Diffuse reflectance spectroscopy has been proven to be a favorable method by which to detect regional [Hb]. This paper introduces a new algorithm to retrieve [Hb] information from diffuse reflectance spectra. The proposed algorithm utilizes the natural logarithmic operation and the differential wavelet transform to effectively quench the scattering effects, and then employs the concept of isosbestic wavelength in the transformed spectra to reduce the effects of hemoglobin oxygenation. As a result, the intensity at the defined isosbestic wavelength of the transformed spectra is a good indicator of [Hb] estimation. The algorithm was derived and validated using theoretical spectra produced by Monte Carlo simulation of photon migration. Its accuracy was further evaluated using liquid tissue phantoms, and its clinical utility with an in vivo clinical study of brain tumors. The results demonstrate the applicability of the algorithm for real-time [Hb] estimations from diffuse reflectance spectra, acquired by means of a fiber-optic spectroscopy system.

Improved detection limits using a hand-held optical imager with coregistration capabilities

Optical imaging is emerging as a non-invasive and non-ionizing method for breast cancer diagnosis. A hand-held optical imager has been developed with coregistration facilities towards flexible imaging of different tissue volumes and curvatures in near real-time. Herein, fluorescence-enhanced optical imaging experiments are performed to demonstrate deeper target detection under perfect and imperfect (100:1) uptake conditions in (liquid) tissue phantoms and in vitro. Upon summation of multiple scans (fluorescence intensity images), fluorescent targets are detected at greater depths than from single scan alone.

Determination of the optical properties of turbid media by measurements of the spatially resolved reflectance considering the point-spread function of the camera system

The radial dependence of the diffuse reflectance from a turbid medium that is due to a point source is basically influenced by the absorption and reduced scattering coefficients. A system consisting of a HeNe laser source and a CCD camera is described for making remote measurements of the spatially resolved diffuse reflectance. Liquid tissue phantoms were made of Intralipid and trypan blue to validate the experimental setup. We show that for the correct determination of the optical properties of the tissue phantoms, the point-spread function (PSF) of the camera system has to be considered. Convolving the PSF with a solution of the diffusion equation, the absorption and reduced scattering coefficients of the tissue phantoms could be determined with an average error of 8% in the absorption coefficient and 4% in the reduced scattering coefficient, whereas without convolution, the errors were considerably larger, especially for large optical parameters.

Variation of haemoglobin extinction coefficients can cause errors in the determination of haemoglobin concentration measured by near-infrared spectroscopy

Near-infrared spectroscopy or imaging has been extensively applied to various biomedical applications since it can detect the concentrations of oxyhaemoglobin (HbO2), deoxyhaemoglobin (Hb) and total haemoglobin (Hbtotal) from deep tissues. To quantify concentrations of these haemoglobin derivatives, the extinction coefficient values of HbO2 and Hb have to be employed. However, it was not well recognized among researchers that small differences in extinction coefficients could cause significant errors in quantifying the concentrations of haemoglobin derivatives. In this study, we derived equations to estimate errors of haemoglobin derivatives caused by the variation of haemoglobin extinction coefficients. To prove our error analysis, we performed experiments using liquid-tissue phantoms containing 1% Intralipid in a phosphate-buffered saline solution. The gas intervention of pure oxygen was given in the solution to examine the oxygenation changes in the phantom, and 3 mL of human blood was added twice to show the changes in [Hbtotal]. The error calculation has shown that even a small variation (0.01 cm−1 mM−1) in extinction coefficients can produce appreciable relative errors in quantification of Δ[HbO2], Δ[Hb] and Δ[Hbtotal]. We have also observed that the error of Δ[Hbtotal] is not always larger than those of Δ[HbO2] and Δ[Hb]. This study concludes that we need to be aware of any variation in haemoglobin extinction coefficients, which could result from changes in temperature, and to utilize corresponding animal's haemoglobin extinction coefficients for the animal experiments, in order to obtain more accurate values of Δ[HbO2], Δ[Hb] and Δ[Hbtotal] from in vivo tissue measurements.