We present the viability of Raman spectroscopic approach for detection and determination of tattoo ink pigments, and the use of polymer optical skin tissue phantoms as possible tools for regulatory agencies.
The detection of molecules by surface-enhanced Raman spectroscopy (SERS) is dependent on the nanomaterial used to induce the enhancement effect. This depends on a variety of parameters of the substrate such as the metal used for their creation, their shape, size and size distribution, concentration, as well as the parameters of the solution, such as packing of the nanoparticles, the complexity of the sample, the solvent, etc. It is most crucial, that the parameters are kept constant to provide uniformity of the enhancement. this is crucial for the development of SERS as a reliable and quantitative technique for bioanalysis. Here, we have developed the silver-core and gold-shell nanoparticles, to serve as the enhancement material. The fabrication phase involved constant concentrations of chemicals stability of the solution physical parameters like stirring and heating, and differed only in the perturbation of the reagents addition kinetics. These nanoparticles were investigated further with their ability to measure the solutions of 2-naphtalenethiol in DMSO, as model for testing the variability of the signal due to the enhancement and the kinetics of the nanoparticle-sample solution during a routine Raman measurement procedure. The results indicate vast difference in the preference of the 2-naphthalenethiol to come into contact with the nanoparticles and the partial enhancement of DMSO in most cases, with an almost complete by-pass of the solvent and direct detection of the 2-naphthalenethiol in one case. Moreover, the kinetics of the measurement solution, or its stability during measurement, is provided.
Utilization of surface-enhanced Raman spectroscopy as a measurement technique is of particular interest in biodetection due to its superb chemical specificity and high sensitivity. The use of SERS substrates further improve this method by massive enhancement of the molecule Raman spectrum, permitting very low levels of detection. Therefore it is important to seek for new ways to develop reliable substrates, which are quickly and easily manufactured at a low cost. This paper describes the development of a simple and cost-effective substrate for the SERS detection. The substrate is synthesized from a silver ink on the glass, and its utilization for biodetection is shown. The hydrophobicity of the substrate permits the pre-concentration benefit of the drop-coating deposition, by the formation of the coffee-ring. This allows to achieve lower limits of detection, by effectively measuring areas with higher concentration of measured molecules than the initial sample. However, the different properties of the medium, such as the influence of protein types and amounts, may influence the ring formation mechanics, thus effectively changing the pre-concentration of the target analyte.
We have developed a numerical model describing the optical and thermal behavior of optical tissue phantoms upon laser irradiation. According to our previous studies, the phantoms can be used as substitute of real skin from the optical, as well as thermal point of view. However, the thermal parameters are not entirely similar to those of real tissues thus there is a need to develop mathematical model, describing the thermal and optical response of such materials. This will facilitate the correction factors, which would be invaluable in translation between measurements on skin phantom to real tissues, and gave a good representation of a real case application.
Here, we present the model dependent on the data of our optical phantoms fabricated and measured in our previous preliminary study. The ambiguity between the modeling and the thermal measurements depend on lack of accurate knowledge of material's thermal properties and some exact parameters of the laser beam. Those parameters were varied in the simulation, to provide an overview of possible parameters' ranges and the magnitude of thermal response.
A quick, cost-effective method for detection of drugs of abuse in biological fluids would be of great value in healthcare, law enforcement, and home testing applications. The alarming rise in narcotics abuse has led to considerable focus on developing potent and versatile analytical tools that can address this societal problem. While laboratory testing plays a key role in the current detection of drug misuse and the evaluation of patients with drug induced intoxication, these typically require expensive reagents and trained personnel, and may take hours to complete. Thus, a significant unmet need is to engineer a facile method that can rapidly detect drugs with little sample preparation, especially the bound fraction that is typically dominant in the blood stream.
Here we report an approach that combines the exquisite sensitivity of surface enhanced Raman spectroscopy (SERS) and a facile protein tethering mechanism to reliably detect four different classes of drugs, barbiturate, benzodiazepine, amphetamine and benzoylecgonine. The proposed approach harnesses the reliable and specific attachment of proteins to both drugs and nanoparticle to facilitate the enhancement of spectral markers that are sensitive to the presence of the drugs. In conjunction with chemometric tools, we have shown the ability to quantify these drugs lower than levels achievable by existing clinical immunoassays. Through molecular docking simulations, we also probe the mechanistic underpinnings of the protein tethering approach, opening the door to detection of a broad class of narcotics in biological fluids within a few minutes as well as for groundwater analysis and toxin detection.
In brain studies, the function of the cerebrospinal fluid (CSF) awakes growing interest, particularly related to studies of the glymphatic system in the brain, which is connected with the complex system of lymphatic vessels responsible for cleaning the tissues. The CSF is a clear, colourless liquid including water (H2O) approximately with a concentration of 99 %. In addition, it contains electrolytes, amino acids, glucose, and other small molecules found in plasma. The CSF acts as a cushion behind the skull, providing basic mechanical as well as immunological protection to the brain. Disturbances of the CSF circulation have been linked to several brain related medical disorders, such as dementia.
Our goal is to develop an in vivo method for the non-invasive measurement of cerebral blood flow and CSF circulation by exploiting optical and capacitive sensing techniques simultaneously. We introduce a prototype of a wearable probe that is aimed to be used for long-term brain monitoring purposes, especially focusing on studies of the glymphatic system. In this method, changes in cerebral blood flow, particularly oxy- and deoxyhaemoglobin, are measured simultaneously and analysed with the response gathered by the capacitive sensor in order to distinct the dynamics of the CSF circulation behind the skull. Presented prototype probe is tested by measuring liquid flows inside phantoms mimicking the CSF circulation.
KEYWORDS: Raman spectroscopy, Interference (communication), Signal processing, Principal component analysis, Denoising, Spectroscopy, Molecular spectroscopy, Signal to noise ratio, Light scattering, Chemical compounds, Modulation, Signal detection, Electronics
We present an overview of noise sources deteriorating the quality of the recorded biological Raman spectra and the ability to determine the specimen composition. The acquired Raman spectra exhibit intense additive noise components or drifts because of low intensity of the scattered light. Therefore we have to apply expensive or bulky measurement setups to limit their inherent noise or to apply additional signal processing to reduce random components after recording the spectra (e.g., Savitzky-Golay filtering, polynomial approximation, denoising by empirical mode decomposition). We present noise sources generated in detectors of Raman scattered photon stream. We consider the methods of background noise reduction by increasing averaging time when the background noise comprises of white noise and 1/f noise components. We consider how the background noise reduces the accuracy of chemical compounds estimation using Raman spectra and prediction model based on linear (e.g., Principal Component Analysis) or nonlinear (e.g., Support Vector Machine) methods. Finally, we give some remarks about synchronous detection and background noise reduction.
We present an surface-enhanced Raman spectroscopy (SERS) approach for detection of drugs of abuse in whole human blood. We utilize a near infrared laser with 830 nm excitation wavelength in order to reduce the influence of fluorescence on the spectra of blood. However, regular plasmon resonance peak of plasmonic nanoparticles, such as silver or gold fall in a much lower wavelength regime about 400 nm. Therefore, we have shifted the plasmon resonance of nanoparticles to match that of an excitation laser wavelength, by fabrication of the silver-core gold-shell nanoparticles. By combining the laser and plasmon resonance shift towards longer wavelengths we have achieved a great reduction in background fluorescence of blood. Great enhancement of Raman signal coming solely from drugs was achieved without any prominent lines coming from the erythrocytes. We have applied chemometric processing methods, such as Principal Component Analysis (PCA), to detect the elusive differences in the Raman bands which are specific for the investigated drugs. We have achieved good classification for the samples containing particular drugs (e.g., butalbital, α-hydroxyalprazolam). Furthermore, a quantitative analysis was carried out to assess the limit of detection (LOD) using Partial Least Squares (PLS) regression method. In conclusion, our LOD values obtained for each class of drugs was competitive with the gold standard GC/MS method.
Human tissue is one of the most complex optical media since it is turbid and nonhomogeneous. In our poster, we suggest a new type of skin phantom and an optical method for sensing physiological tissue condition, basing on the collection of the ejected light at all exit angles, to receive the full scattering profile. Conducted experiments were carried out on an unique set-up for noninvasive encircled measurement. Set-up consisted of a laser, a photodetector and new tissues-like phantoms made with a polyvinyl chloride-plastisol (PVCP), silicone elastomer polydimethylsiloxane (PDMS) and PDMS with glycerol mixture. Our method reveals an isobaric point, which is independent of the optical properties. Furthermore, we present the angular distribution of cylindrical phantoms, in order to sense physiological tissue state.
A wide range of dermatological diseases can be efficiently treated using laser heating. Nevertheless, before the new laser
is introduced into clinical practice, its parameters and ability to interact with human skin have to be carefully examined.
In order to do that optical skin phantoms can be used. Such phantoms closely imitate the scattering and absorption
properties of real human skin tissue along with its thermal properties, such as capacitance and conductivity specific heat.
We have fabricated a range of optical tissue phantoms based on polyvinylchloride-plastisol PVC-P with varying optical
properties, including the absorption, scattering and density of the matrix material. We have utilized a pre-clinical
dermatological laser system with a 975 nm diode laser module. A range of laser settings were tested, such as laser pulse
duration, laser power and number of pulses. We have studied laser irradiation efficiency on fabricated optical tissue
phantoms. Measurements of the temporal and spatial temperature distribution on the phantoms' surface were performed
using thermographic imaging. The comparison of results between tissues' and phantoms' optical and thermal response
prove that they can be used for approximate evaluation of laser heating efficiency. This study presents a viable approach
for calibration of dermatological lasers which can be utilized in practice.
We present optoelectronic investigation of in vitro interactions of whole human blood with different nanodiamond biomarkers. Plasmo-chemical modifications of detonation nanodiamond particles gives the possibility for controlling their surface for biological applications. Optical investigations reveal the biological activity of nanodiamonds in blood dependent on its surface termination. We compare different types of nanodiamonds: commercial non-modified detonation nanodiamonds, and nanodiamonds modified by MW PACVD method with H2-termination, and chemically modified nanodiamond with O2-termination. The absorption spectra, and optical microscope investigations were conducted. The results indicate haemocompatibility of non-modified detonation nanodiamond as well as modified nanodiamonds, which enables their application for drug delivery, as well as sensing applications.
A dedicated absorption spectroscopy system was set up using tungsten-halogen broadband source, optical fibers, sample holder, and a commercial spectrometer with CCD array. Analysis of noise present in the setup was carried out. Data processing was applied to the absorption spectra to reduce spectral noise, and improve the quality of the spectra and to remove the baseline level. The absorption spectra were measured for whole blood samples, separated components: plasma, saline, washed erythrocytes in saline and human whole blood with biomarkers - biocompatible nanodiamonds (ND). Blood samples had been derived from a number of healthy donors. The results prove a correct setup arrangement, with adequate preprocessing of the data. The results of blood-ND mixtures measurements show no toxic effect on blood cells, which proves the NDs as a potential biocompatible biomarkers.
Skin lesions are commonly treated using laser heating. However, the introduction of new devices into clinical practice requires evaluation of their performance. This study presents the application of optical phantoms for assessment of a newly developed 975-nm pulsed diode laser system for dermatological purposes. Such phantoms closely mimic the absorption and scattering of real human skin (although not precisely in relation to thermal conductivity and capacitance); thus, they can be used as substitutes for human skin for approximate evaluation of laser heating efficiency in an almost real environment. Thermographic imaging was applied to measure the spatial and temporal temperature distributions on the surface of laser-irradiated phantoms. The study yielded results of heating with regard to phantom thickness and absorption, as well as laser settings. The methodology developed can be used in practice for preclinical evaluations of laser treatment for dermatology.
Our aim is to optically monitor the delivery of the chemotherapy drugs for brain tumours, particularly used in the central nervous system (CNS) lymphoma therapy. In vivo monitoring would help to optimize the treatment and avoiding unnecessary medications. Moreover, it would be beneficial to be able to measure which of the multi-regimen drugs actually do penetrate and how well into the brain tissue.
There exist several potential optical measurement techniques to be utilised for the purpose. The most desired method would allow the detection of the drugs without using optical biomarkers as a contrast agent. In this case, for non-invasive sensing of the drug in the brain cortex, the drug should have a reasonably strong optical absorption band somewhere in the range between 600 nm and 1700 nm, and not directly coincident with the strong bands of haemoglobin or water. Alternatively, mid-infrared (MIR) range has the potential for invasive drug monitoring techniques.
In this paper, we report the optical properties of several chemotherapy drugs used in CNS lymphoma therapy, such as rituximabi, cyclophosphamide and etoposide. We measured their transmittance and reflectance spectra in near-infrared (NIR) range, particularly 900 nm − 2500 nm, to be considered when choosing the in vivo monitoring method to be developed. The absorption and scattering coefficients were retrieved from the measurements and applying Beer’s law. For the measurement of the sum of total transmission and reflection in NIR range we used integrating sphere with spektralo to enable calculation of the scattering coefficient.
In this article the simultaneous investigation of blood parameters by complementary optical methods, Raman spectroscopy and spectral-domain low-coherence interferometry, is presented. Thus, the mutual relationship between chemical and physical properties may be investigated, because low-coherence interferometry measures optical properties of the investigated object, while Raman spectroscopy gives information about its molecular composition.
A series of in-vitro measurements were carried out to assess sufficient accuracy for monitoring of blood parameters. A vast number of blood samples with various hematological parameters, collected from different donors, were measured in order to achieve a statistical significance of results and validation of the methods. Preliminary results indicate the benefits in combination of presented complementary methods and form the basis for development of a multimodal system for rapid and accurate optical determination of selected parameters in whole human blood. Future development of optical systems and multivariate calibration models are planned to extend the number of detected blood parameters and provide a robust quantitative multi-component analysis.
KEYWORDS: Blood, Raman spectroscopy, Data modeling, Spectroscopy, Plasma, Neodymium, Statistical analysis, In vivo imaging, Quantitative analysis, Chemical analysis
In this paper we present a proof-of-concept of a Raman spectroscopy-based approach for measuring the content of propofol, a common anesthesia drug, in whole human blood, and plasma, which is intended for use during clinical procedures. This method utilizes the Raman spectroscopy as a chemically–sensitive method for qualitative detection of the presence of a drug and a quantitative determination of its concentration. A number of samples from different patients with added various concentrations of propofol IV solution were measured. This is most equivalent to a real in-vivo situation. Subsequent analysis of a set of spectra was carried out to extract qualitative and quantitative information. We conclude, that the changes in the spectra of blood with propofol, overlap with the most prominent lines of the propofol solution, especially at spectral regions: 1450 cm-1, 1250- 1260 cm-1, 1050 cm-1, 875-910 cm-1, 640 cm-1. Later, we have introduced a quantitative analysis program based on correlation matrix closest fit, and a LOO cross-validation. We have achieved 36.67% and 60% model precision when considering full spectra, or specified bands, respectively. These results prove the possibility of using Raman spectroscopy for quantitative detection of propofol concentrations in whole human blood.
We present the optical measurement techniques used in human skin phantom studies. Their accuracy and the sources of errors in microscopic parameters’ estimation of the produced phantoms are described. We have produced optical phantoms for the purpose of simulating human skin tissue at the wavelength of 930 nm. Optical coherence tomography was used to measure the thickness and surface roughness and to detect the internal inhomogeneities. A more detailed study of phantom surface roughness was carried out with the optical profilometer. Reflectance, transmittance, and collimated transmittance of phantoms were measured using an integrating-sphere spectrometer setup. The scattering and absorption coefficients were calculated with the inverse adding-doubling method. The reduced scattering coefficient at 930 nm was found to be 1.57±0.14 mm−1 and the absorption was 0.22±0.03 mm−1. The retrieved optical properties of phantoms are in agreement with the data found in the literature for real human tissues.
This paper reports preliminary studies on use of Raman spectroscopy for investigation of blood. High quality blood spectra were recorded in-vitro with excitation wavelengths of 830 nm. Because of complex composition of the blood as well as by light attenuation and scattering in the tissues, spectra set up from wide, low-intensive Raman bands and intensive optical background. To get information about origin of bands in Raman spectra it is necessary to create phantom, which would show influence of this parameter and can be used to calibrate the Raman measurement system. Spectra of phantoms of selected blood components were acquired and discussed.
Dermatological diseases, such as neurofibroma (Recklinghausen disease) or hemangiomas can be efficiently treated using photothermolysis from laser irradiation. We have utilized a developed 975 nm fiber diode laser as a low-cost alternative over common Nd:YAG lasers. This paper describes the investigations of interaction of 975 nm diode laser radiation-pulses with optical skin phantoms which were designed and manufactured in our laboratory. Such phantoms match the scattering and absorption coefficients of real human skin. Spatial and temporal temperature evolutions during laser irradiation with various laser settings (pulsed and CW mode), were recorded by an IR camera. Subsequent analysis yielded optimum choice of parameters for laser therapy of coetaneous lesions.
In this article the procedure of selection of physiological parameters for optoelectronic system supporting behavioral
therapy of autistic children is proposed. Authors designed and conducted an experiment in which a group of 30 health volunteers (16 females and 14 males) were examined. Under controlled conditions people were exposed to a stressful situation caused by the picture or sound (1kHz constant sound, which was gradually silenced and finished with a shot sound). For each of volunteers, a set of physiological parameters were recorded, including: skin conductance, heart rate,
peripheral temperature, respiration rate and electromyography. The selected characteristics were measured in different
locations in order to choose the most suitable one for the designed therapy supporting system. The bio-statistical analysis allowed us to discern the proper physiological parameters that are most associated to changes due to emotional state of a
patient, such as: skin conductance, temperatures and respiration rate. This allowed us to design optoelectronic sensors
network for supporting behavioral therapy of children with autism.
We present the implementation and validation of low-coherence Fabry–Perot interferometer for refractive index dispersion measurements of liquids. A measurement system has been created with the use of four superluminescent diodes with different optical parameters, a fiber-optic coupler and an optical spectrum analyzer. The Fabry–Perot interferometer cavity has been formed by the fiber-optic end and mirror surfaces mounted on a micromechanical stage. The positive result of the validation procedure has been determined through statistical analysis. All obtained results were 99.999% statistically significant and were characterized by a strong positive correlation (r>0.98). The accuracy of the measured result of implemented low-coherence Fabry–Perot interferometer sensor is from 83% to 94%, which proves that the sensor can be used in the measurement of refractive index dispersion of liquids.
Reliability and validity of measurements is of utmost importance when assessing measuring capability of instruments developed for research. In order to perform an experiment which is legitimate, used instruments must be both reliable and valid. Reliability estimates the degree of precision of measurement, the extent to which a measurement is internally consistent. Validity is the usefulness of an instrument to perform accurate measurements of quantities it was designed to measure. Statistical analysis for reliability and validity control of low-coherence interferometry method for refractive index measurements of biological fluids is presented. The low-coherence interferometer is sensitive to optical path difference between interfering beams. This difference depends on the refractive index of measured material. To assess the validity and reliability of proposed method for blood measurements, the statistical analysis of the method was performed on several substances with known refractive indices. Analysis of low-coherence interferograms considered the mean distances between fringes. Performed statistical analysis for validity and reliability consisted of Grubb’s test for outliers, Shapiro-Wilk test for normal distribution, T-Student test, standard deviation, coefficient of determination and r-Pearson correlation. Overall the tests proved high statistical significance of measurement method with confidence level < 0.0001 of measurement method.
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