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<p>Filter-Membrane-Based Ultrafiltration Coupled with Surface-Enhanced Raman Spectroscopy for Potential Differentiation of Benign and Malignant Thyroid Tumors from Blood Plasma</p>
摘要： The objective of this study is to evaluate the performance and feasibility of surface-enhanced Raman spectroscopy coupled with a filter membrane and advanced multivariate data analysis on identifying and differentiating benign and malignant thyroid tumors from blood plasma. We proposed a membrane filter SERS technology for the differentiation between benign thyroid tumor and thyroid cancer. That is to say, by using filter membranes with optimal pore size, the blood plasma samples from thyroid tumor patients were pretreated with the macromolecular proteins being filtered out prior to SERS measurement. The SERS spectra of blood plasma ultrafiltrate obtained using filter membranes from 102 patients with thyroid tumors (70 thyroid cancers and 32 benign thyroid tumors) were then analyzed and compared. Two multivariate statistical analyses, principal component analysis-linear discriminate and Lasso-partial least squares-discriminant analysis (Lasso-PLS-DA), were performed on the SERS spectral data after background subtraction and normalization, as well as the first derivative processing, to analyze and compare the differential diagnosis of benign thyroid tumors and thyroid cancer. SERS measurements were performed in blood plasma acquired from a total of 102 thyroid tumor patients (benign thyroid tumor N=32; thyroid cancer N=70). By using filter membranes, the macromolecular proteins in blood plasma were effectively filtered out to yield high-quality SERS spectra. 84.3% discrimination accuracy between benign and malignant thyroid tumor was achieved using PCA-LDA method, while Lasso-PLS-DA yields a discrimination accuracy of 90.2%. Our results demonstrate that SERS spectroscopy, coupled with ultrafiltration and multivariate analysis has the potential of providing a non-invasive, rapid, and objective detection and differentiation of benign and malignant thyroid tumors.
关键词： thyroid tumor,silver nanoparticles,filter membrane,blood plasma,surface-enhanced Raman spectroscopy
更新于2025-09-19 17:13:59
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Cost-effective microfabrication of sub-micron-depth channels by femto-laser anti-stiction texturing
摘要： Micro Electro Mechanical Systems (MEMS) and microfluidic devices have found numerous applications in the industrial sector. However, they require a fast, cost-effective and reliable manufacturing process in order to compete with the conventional methods. Particularly, at the sub-micron scale, the manufacturing od devices are limited by the dimensional complexity. A proper bonding and stiction prevention of these sub-micron channels are two of the main challenges faced during the fabrication process of low aspect ratio channels. Especially, in case of using flexible materials such as polydimethylsiloxane (PDMS). This study presents a direct laser microfabrication method of sub-micron channels using an infrared (IR) ultrashort pulse (femtoseconds) capable of manufacturing extremely low aspect ratio channels. These microchannels are manufactured and tested varying their depth from 0.5 μm to 2 μm and width of 15, 20, 25, and 30 μm. The roughness of each pattern was measured by an interferometric microscope. Additionally, the static contact angle of each depth was studied to evaluate the influence of femtosecond laser fabrication method on the wettability of the glass substrate. PDMS, which is a biocompatible polymer, was used to provide a watertight property to the sub-micron channels and also to assist the assembly of external microfluidic hose connections. A 750nm depth watertight channel was built using this methodology and successfully used as a blood plasma separator (BPS). The device was able to achieve 100% pure plasma without stiction of the PDMS layer to the sub-micron channel within an adequate time. This method provides a novel manufacturing approach useful for various applications such as point-of-care devices.
关键词： Femtolaser,Biofabrication,Microfluidic,Blood/plasma separation
更新于2025-09-16 10:30:52
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Autofluorescence of Breast Cancer Proteins
摘要： Background: The intensity of the blood as well as tissue autofluorescence (fluorescence of endogenous fluorophores) shows current structure of protein mixture, its current conformation (native or denatured state), concentration or activity depending on the external and internal conditions. Nicotine amide adenine dinucleotide (NAD+) is a cofactor in redox reactions of glycolysis and the citric acid cycle in eukaryotic cells and it can be a marker of the intensity of mitochondria metabolism as well as the presence of oxygen in the cells. The concentration of reduced nicotinamide adenine dinucleotide (NADH) varies during normoxia, hypoxia and hyperoxia cells. Cancer cells increase the concentration of NADH during hypoxia and anoxia. The reason of NADH cumulation is microenvironment with low or no oxygen supply which induces glycolysis as a preferential source of energy. Glycolysis is faster, does not need oxygen but is less effective than oxidative phosphorylation. Focus: The aim of this work was to measure the structure of blood plasma and mammary gland homogenates of patients at three stages of breast cancer in comparison to healthy subjects using fluorescence analysis and atomic force microscopy. The blood plasma of patients with breast cancer had a different structure of proteins in comparison to healthy subjects. The blood plasma and homogenate of patients with breast cancer showed significant increase in autofluorescence intensity, which represents mixture of various endogenous fluorophores in particular porphyrins, collagen, NAD and flavins. Prospect: The information about complex summary of fluorescence intensity of all mixtures of endogenous fluorophores may in the future serve as rapid preliminary markers of cancer. The fluorescence analysis might be a non-traditional methodology for an early rapid diagnosis of breast cancer in the next clinical practice.
关键词： breast cancer proteins,blood plasma,Nicotine amide adenine dinucleotide,breast cancer homogenate,fluorophores,autofluorescence
更新于2025-09-04 15:30:14
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In-line whole blood fractionation for Raman analysis of blood plasma
摘要： Blood plasma evaluation has high significance in clinical diagnostics. Current schemes involve the preparation of blood plasma by centrifugation of whole blood followed by electrochemical or spectroscopic analysis. However, centrifugation is often too time-consuming for application in clinical emergency and point-of-care settings. We propose to combine microfluidic, instantaneous plasma fractionation with localized spectroscopic methods for in-line analysis. As an example, we present confocal Raman spectroscopy in fractionated plasma domains at two different Raman excitation wavelengths. Resonance Raman spectroscopy with laser excitation at 408 nm allows the specific detection of free hemoglobin in blood plasma at concentrations above 22 mg dl?1 (level of detection). Consequently, we are able to accurately resolve the range of clinical relevance regarding hemolysis. At near-infrared excitation (785 nm) we furthermore demonstrate the acquisition of characteristic Raman spectra of fractionated blood plasma in the microfluidic setting. These spectra can serve as starting point for a multi-parameter regression analysis to quantify a set of blood plasma parameters from a single Raman spectrum. The combined microfluidics and Raman spectroscopy method is non-destructive and has a whole blood consumption of less than 100 μl per hour. It thus allows for continuous in-line blood plasma monitoring.
关键词： microfluidics,hemoglobin,Raman spectroscopy,blood plasma,clinical diagnostics
更新于2025-09-04 15:30:14