Characterization and classification of ductal carcinoma tissue using four channel based stokes-mueller polarimetry and machine learning.

Interaction of polarized light with healthy and abnormal regions of tissue reveals structural information associated with its pathological condition. Even a slight variation in structural alignment can induce a change in polarization property, which can play a crucial role in the early detection of abnormal tissue morphology. We propose a transmission-based Stokes-Mueller microscope for quantitative analysis of the microstructural properties of the tissue specimen. The Stokes-Mueller based polarization microscopy provides significant structural information of tissue through various polarization parameters such as degree of polarization (DOP), degree of linear polarization (DOLP), and degree of circular polarization (DOCP), anisotropy (r) and Mueller decomposition parameters such as diattenuation, retardance and depolarization. Further, by applying a suitable image processing technique such as Machine learning (ML) output images were analysed effectively. The support vector machine image classification model achieved 95.78% validation accuracy and 94.81% testing accuracy with polarization parameter dataset. The study's findings demonstrate the potential of Stokes-Mueller polarimetry in tissue characterization and diagnosis, providing a valuable tool for biomedical applications.

Skin emitted volatiles analysis for noninvasive diagnosis: the current advances in sample preparation techniques for biomedical application.

Human skin emits a series of volatile compounds from the skin due to various metabolic processes, microbial activity, and several external factors. Changes in the concentration of skin volatile metabolites indicate many diseases, including diabetes, cancer, and infectious diseases. Researchers focused on skin-emitted compounds to gain insight into the pathophysiology of various diseases. In the case of skin volatolomics research, it is noteworthy that sample preparation, sampling protocol, analytical techniques, and comprehensive validation are important for the successful integration of skin metabolic profiles into regular clinical settings. Solid-phase microextraction techniques and polymer-based active sorbent traps were developed to capture the skin-emitted volatile compounds. The primary advantage of these sample preparation techniques is the ability to efficiently and targetedly capture skin metabolites, thus improving the detection of the biomarkers associated with various diseases. In further research, polydimethyl-based patches were utilized for skin research due to their biocompatibility and thermal stability properties. The microextraction sampling tools coupled with high sensitive Gas Chromatography-Mass Spectrometer provided a potential platform for skin volatolomes, thus emerging as a state-of-the-art analytical technique. Later, technological advancements, including the design of wearable sensors, have enriched skin-based research as it can integrate the information from skin-emitted volatile profiles into a portable platform. However, individual-specific hydration, temperature, and skin conditions can influence variations in skin volatile concentration. Considering the subject-specific skin depth, sampling time standardization, and suitable techniques may improve the skin sampling techniques for the potential discovery of various skin-based marker compounds associated with diseases. Here, we have summarised the current research progress, limitations, and technological advances in skin-based sample preparation techniques for disease diagnosis, monitoring, and personalized healthcare applications.

Resistive Memory-Switching Behavior in Solution-Processed Trans, trans-1,4-bis-(2-(2-naphthyl)-2-(butoxycarbonyl)-vinyl) Benzene-PVA-Composite-Based Aryl Acrylate on ITO-Coated PET.

Resistive switching memories are among the emerging next-generation technologies that are possible candidates for in-memory and neuromorphic computing. In this report, resistive memory-switching behavior in solution-processed trans, trans-1,4-bis-(2-(2-naphthyl)-2-(butoxycarbonyl)-vinyl) benzene-PVA-composite-based aryl acrylate on an ITO-coated PET device was studied. A sandwich configuration was selected, with silver (Ag) serving as a top contact and trans, trans-1,4-bis-(2-(2-naphthyl)-2-(butoxycarbonyl)-vinyl) benzene-PVA-composite-based aryl acrylate and ITO-PET serving as a bottom contact. The current-voltage (I-V) characteristics showed hysteresis behavior and non-zero crossing owing to voltages sweeping from positive to negative and vice versa. The results showed non-zero crossing in the devices' current-voltage (I-V) characteristics due to the nanobattery effect or resistance, capacitive, and inductive effects. The device also displayed a negative differential resistance (NDR) effect. Non-volatile storage was feasible with non-zero crossing due to the exhibition of resistive switching behavior. The sweeping range was -10 V to +10 V. These devices had two distinct states: 'ON' and 'OFF'. The ON/OFF ratios of the devices were 14 and 100 under stable operating conditions. The open-circuit voltages (Voc) and short-circuit currents (Isc) corresponding to memristor operation were explained. The DC endurance was stable. Ohmic conduction and direct tunneling mechanisms with traps explained the charge transport model governing the resistive switching behavior. This work gives insight into data storage in terms of a new conception of electronic devices based on facile and low-temperature processed material composites for emerging computational devices.

Exploring the future of regenerative medicine: Unveiling the potential of optical microscopy for structural and functional imaging of stem cells.

Regenerative medicine, which utilizes stem cells for tissue and organ repair, holds immense promise in healthcare. A comprehensive understanding of stem cell characteristics is crucial to unlock their potential. This study explores the pivotal role of optical microscopy in advancing regenerative medicine as a potent tool for stem cell research. Advanced optical microscopy techniques enable an in-depth examination of stem cell behavior, morphology, and functionality. The review encompasses current optical microscopy, elucidating its capabilities and constraints in stem cell imaging, while also shedding light on emerging technologies for improved stem cell visualization. Optical microscopy, complemented by techniques like fluorescence and multiphoton imaging, enhances our comprehension of stem cell dynamics. The introduction of label-free imaging facilitates noninvasive, real-time stem cell monitoring without external dyes or markers. By pushing the boundaries of optical microscopy, researchers reveal the intricate cellular mechanisms underpinning regenerative processes, thereby advancing more effective therapeutic strategies. The current study not only outlines the future of regenerative medicine but also underscores the pivotal role of optical microscopy in both structural and functional stem cell imaging.

Isolation, Detection and Analysis of Circulating Tumour Cells: A Nanotechnological Bioscope.

Cancer is one of the dreaded diseases to which a sizeable proportion of the population succumbs every year. Despite the tremendous growth of the health sector, spanning diagnostics to treatment, early diagnosis is still in its infancy. In this regard, circulating tumour cells (CTCs) have of late grabbed the attention of researchers in the detection of metastasis and there has been a huge surge in the surrounding research activities. Acting as a biomarker, CTCs prove beneficial in a variety of aspects. Nanomaterial-based strategies have been devised to have a tremendous impact on the early and rapid examination of tumor cells. This review provides a panoramic overview of the different nanotechnological methodologies employed along with the pharmaceutical purview of cancer. Initiating from fundamentals, the recent nanotechnological developments toward the detection, isolation, and analysis of CTCs are comprehensively delineated. The review also includes state-of-the-art implementations of nanotechnological advances in the enumeration of CTCs, along with future challenges and recommendations thereof.

Types of spectroscopy and microscopy techniques for cancer diagnosis: a review.

Cancer is a life-threatening disease that has claimed the lives of many people worldwide. With the current diagnostic methods, it is hard to determine cancer at an early stage, due to its versatile nature and lack of genomic biomarkers. The rapid development of biophotonics has emerged as a potential tool in cancer detection and diagnosis. Using the fluorescence, scattering, and absorption characteristics of cells and tissues, it is possible to detect cancer at an early stage. The diagnostic techniques addressed in this review are highly sensitive to the chemical and morphological changes in the cell and tissue during disease progression. These changes alter the fluorescence signal of the cell/tissue and are detected using spectroscopy and microscopy techniques including confocal and two-photon fluorescence (TPF). Further, second harmonic generation (SHG) microscopy reveals the morphological changes that occurred in non-centrosymmetric structures in the tissue, such as collagen. Again, Raman spectroscopy is a non-destructive method that provides a fingerprinting technique to differentiate benign and malignant tissue based on Raman signal. Photoacoustic microscopy and spectroscopy of tissue allow molecule-specific detection with high spatial resolution and penetration depth. In addition, terahertz spectroscopic studies reveal the variation of tissue water content during disease progression. In this review, we address the applications of spectroscopic and microscopic techniques for cancer detection based on the optical properties of the tissue. The discussed state-of-the-art techniques successfully determines malignancy to its rapid diagnosis.

Gamma radiation as a modifier of starch - Physicochemical perspective.

Starch is one of the most common and abundantly found carbohydrates in cereals, roots, legumes, and some fruits. It is a tasteless, colorless, and odorless source of energy that is present in the amyloplasts of plants. Native starch comprises amylose, a linear α-glucan having α-1,4-linkage and amylopectin, a branched polysaccharide with both α-1,4-linkage and α-1,6-linkage. Due to the low solubility, high viscosity, and unstable pasting property of native starch, it has been restricted from its application in industries. Although native starch has been widely used in various industries, modification of the same by various chemical, enzymatic and physical methods have been carried out to alter its properties for better performance in several industrial aspects. Physical modification like gamma radiation is frequently used as it is rapid, penetrates deeper, less toxic, and cost-effective. Starch when irradiated with gamma rays is observed to produce free radicals, generate sugars owing to cleavage of amylopectin branches, and exhibit variation in enzymatic digestion, amylose content, morphology, crystallinity, thermal property, and chemical composition. These physicochemical properties of the starch due to gamma radiation are assessed using optical microscopy, scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier-transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), and its application are discussed.

Elucidating Methods for Isolation and Quantification of Exosomes: A Review.

Exosomes are the smallest extracellular vesicles present in most of the biological fluids. They are found to play an important role in cell signaling, immune response, tumor metastasis, etc. Studies have shown that these vesicles also have diagnostic and therapeutic roles for which their accurate detection and quantification is essential. Due to the complexity in size and structure of exosomes, even the gold standard methods face challenges. This comprehensive review discusses the various standard methods such as ultracentrifugation, ultrafiltration, size-exclusion chromatography, precipitation, immunoaffinity, and microfluidic technologies for the isolation of exosomes. The principle of isolation of each method is described, as well as their specific advantages and disadvantages. Quantification of exosomes by nanoparticle tracking analysis, flow cytometry, tunable resistive pulse sensing, electron microscopy, dynamic light scattering, and microfluidic devices are also described, along with the applications of exosomes in various biomedical domains.

Photodynamic therapy to control microbial biofilms.

Microorganisms thrive in well-organized biofilm ecosystems. Biofilm-associated cells typically show increased resistance to antibiotics and contribute significantly to treatment failure. This has prompted investigations aimed at developing advanced and novel antimicrobial approaches that could effectively overcome the shortcomings associated with conventional antibiotic therapy. Studies are ongoing to develop effective curative strategies ranging from the use of peptides, small molecules, nanoparticles to bacteriophages, sonic waves, and light energy targeting various structural and physiological aspects of biofilms. In photodynamic therapy, a light source of a specific wavelength is used to irradiate non-toxic photosensitizers such as tetrapyrroles, synthetic dyes or, naturally occurring compounds to generate reactive oxygen species that can exert a lethal effect on the microbe especially by disrupting the biofilm. The photosensitizer preferentially binds to and accumulates in the microbial cells without causing any damage to the host tissue. Currently, photodynamic therapy is increasingly being used for the treatment of oral caries and dental plaque, chronic wound infections, infected diabetic foot ulcers, cystic fibrosis, chronic sinusitis, implant device-associated infections, etc. This approach is recognized as safe, as it is non-toxic and minimally invasive, making it a reliable, realistic, and promising therapeutic strategy for reducing the microbial burden and biofilm formation in chronic infections. In this review article, we discuss the current and future potential strategies of utilizing photodynamic therapy to extend our ability to impede and eliminate biofilms in various medical conditions.

Recent trends in two-photon auto-fluorescence lifetime imaging (2P-FLIM) and its biomedical applications.

Two photon fluorescence microscopy and the numerous technical advances to it have served as valuable tools in biomedical research. The fluorophores (exogenous or endogenous) absorb light and emit lower energy photons than the absorption energy and the emission (fluorescence) signal is measured using a fluorescence decay graph. Additionally, high spatial resolution images can be acquired in two photon fluorescence lifetime imaging (2P-FLIM) with improved penetration depth which helps in detection of fluorescence signal in vivo. 2P-FLIM is a non-invasive imaging technique in order to visualize cellular metabolic, by tracking intrinsic fluorophores present in it, such as nicotinamide adenine dinucleotide, flavin adenine dinucleotide and tryptophan etc. 2P-FLIM of these molecules enable the visualization of metabolic alterations, non-invasively. This comprehensive review discusses the numerous applications of 2P-FLIM towards cancer, neuro-degenerative, infectious diseases, and wound healing.

Polarization-resolved Stokes-Mueller imaging: a review of technology and applications.

Polarization microscopy, a powerful optical tool to study anisotropic properties of biomolecules, provides better microstructural information of a sample as compared with conventional optical microscopic techniques. The measurement and analysis of polarization states of light can be performed using both Jones matrix as well as Stokes algebra. Further, the details of optical properties of specimen are characterized by Mueller matrix. However, the application of Jones calculus is limited to perfectly polarized light, but Stokes-Mueller polarimetry is emerging as a promising tool for tissue imaging due to its application irrespective of polarization state of the light. In this review article, we explain the development of Stokes-Mueller formalism in context of linear optics. Furthermore, application of Mueller matrix decomposition (MMD) method to derive sample properties is demonstrated in several bio-medical studies.

Types of advanced optical microscopy techniques for breast cancer research: a review.

A cancerous cell is characterized by morphological and metabolic changes which are the key features of carcinogenesis. Adenosine triphosphate (ATP) in cancer cells is primarily produced by aerobic glycolysis rather than oxidative phosphorylation. In normal cellular metabolism, nicotinamide adenine dinucleotide (NADH) is considered as a principle electron donor and flavin adenine dinucleotide (FAD) as an electron acceptor. During metabolism in a cancerous cell, a net increase in NADH is found as the pathway switched from oxidative phosphorylation to aerobic glycolysis. Often during initiation and progression of cancer, the developmental regulation of extracellular matrix (ECM) is restricted and becomes disorganized. Tumor cell behavior is regulated by the ECM in the tumor micro environment. Collagen, which forms the scaffold of tumor micro-environment also influences its behavior. Advanced optical microscopy techniques are useful for determining the metabolic characteristics of cancerous, normal cells and tissues. They can be used to identify the collagen microstructure and the function of NADH, FAD, and lipids in living system. In this review article, various optical microscopy techniques applied for breast cancer research are discussed including fluorescence, confocal, second harmonic generation (SHG), coherent anti-Stokes Raman scattering (CARS), and fluorescence lifetime imaging (FLIM).

Fluorescence lifetime imaging of alterations to cellular metabolism by domain 2 of the hepatitis C virus core protein.

Hepatitis C virus (HCV) co-opts hepatic lipid pathways to facilitate its pathogenesis. The virus alters cellular lipid biosynthesis and trafficking, and causes an accumulation of lipid droplets (LDs) that gives rise to hepatic steatosis. Little is known about how these changes are controlled at the molecular level, and how they are related to the underlying metabolic states of the infected cell. The HCV core protein has previously been shown to independently induce alterations in hepatic lipid homeostasis. Herein, we demonstrate, using coherent anti-Stokes Raman scattering (CARS) microscopy, that expression of domain 2 of the HCV core protein (D2) fused to GFP is sufficient to induce an accumulation of larger lipid droplets (LDs) in the perinuclear region. Additionally, we performed fluorescence lifetime imaging of endogenous reduced nicotinamide adenine dinucleotides [NAD(P)H], a key coenzyme in cellular metabolic processes, to monitor changes in the cofactor's abundance and conformational state in D2-GFP transfected cells. When expressed in Huh-7 human hepatoma cells, we observed that the D2-GFP induced accumulation of LDs correlated with an increase in total NAD(P)H fluorescence and an increase in the ratio of free to bound NAD(P)H. This is consistent with an approximate 10 fold increase in cellular NAD(P)H levels. Furthermore, the lifetimes of bound and free NAD(P)H were both significantly reduced--indicating viral protein-induced alterations in the cofactors' binding and microenvironment. Interestingly, the D2-expressing cells showed a more diffuse localization of NAD(P)H fluorescence signal, consistent with an accumulation of the co-factor outside the mitochondria. These observations suggest that HCV causes a shift of metabolic control away from the use of the coenzyme in mitochondrial electron transport and towards glycolysis, lipid biosynthesis, and building of new biomass. Overall, our findings demonstrate that HCV induced alterations in hepatic metabolism is tightly linked to alterations in NAD(P)H functional states.

7-Ketocholesterol induces P-glycoprotein through PI3K/mTOR signaling in hepatoma cells.

7-Ketocholesterol (7-KC) is found at an elevated level in patients with cancer and chronic liver disease. The up-regulation of an efflux pump, P-glycoprotein (P-gp) leads to drug resistance. To elucidate the effect of 7-KC on P-gp, P-gp function and expression were investigated in hepatoma cell lines Huh-7 and HepG2 and in primary hepatocyte-derived HuS-E/2 cells. At a subtoxic concentration, 48-h exposure to 7-KC reduced the intracellular accumulation and cytotoxicity of P-gp substrate doxorubicin in hepatoma cells, but not in HuS-E/2 cells. In Huh-7 cells, 7-KC elevated efflux function through the activation of phosphatidylinositol 3-kinase (PI3K)/mammalian target of rapamycin (mTOR) pathway. 7-KC activated the downstream protein synthesis initiation factor 4E-BP1 and induced P-gp expression post-transcriptionally. The stimulation of efflux was reversible and could not be prevented by N-acetyl cysteine. Total cellular ATP content remained the same, whereas the lactate production was increased and fluorescence lifetime of protein-bound NADH was shortened. These changes suggested a metabolic shift to glycolysis, but glycolytic inhibitors did not eliminate 7-KC-mediated P-gp induction. These results demonstrate that 7-KC induces P-gp through PI3K/mTOR signaling and decreased the cell-killing efficacy of doxorubicin in hepatoma cells.