Bodipy-based quinoline derivative as a highly Hg2+-selective fluorescent chemosensor and its potential applications.

A highly efficient sensor has been successfully developed using quinoline-based BODIPY compounds (8-quinoline-4,4-difluoro-4-boro-3a, 4a-diazaindacene (C1) and 7-hydroxy-8-quinoline-4,4-difluoro-4-boro-3a, 4a-diazindacene (C2) to detect Hg2+ ions. The sensor C1 exhibits remarkable selectivity in detecting Hg2+ with a limit of detection 3.06 × 10-8 mol/L. The developed chemical sensors have shown stability, cost-effectiveness, ease of preparation, and remarkable selectivity towards Hg2+ ions compared to other commonly occurring metal ions. The total recovery of the sensor C1 can be achieved by using a 0.1 mol/L solution of KI. The proposed sensor C1 has been applied to determine Hg2+ in tap and distilled water, yielding excellent results. In addition, the binding mode of C1-Hg2+ and C2-Hg2+ complexes was a 1:1 ratio confirmed by mass spectra, Job's plot, and DFT study. Moreover, the sensor C1 successfully applied for the biological studies results in negligible cytotoxicity, which demonstrates it can be used to determine Hg2+ in HT22 cells.

A fluorescence turn "on-off" imaging probe for sequential detection of Al3+ and L-Cysteine in HeLa cells.

At this "Aluminum Age", exposure to aluminum (metallic or ionic form) is inevitable and inestimable. The presence of aluminum in biological systems is evident but more often aluminum toxicity is less understood. Therefore, the presence of biologically reactive aluminum needs to be identified and quantified. Alongside metals, L-cysteine, an essential amino acid, plays a pivotal role in the homeostasis of cellular oxidative and reductive stress. However, excess (<7g) could be lethal and can lead to death. Thus, in-situ selective detection of aluminum and L-cysteine is of larger interest. Here we report a fluorogenic probe (R) for the sequential selective detection and quantification of Al3+ and L-cysteine in a semi-aqueous medium (3:7; water: DMSO). The probe (R) was synthesized by a one-step acid-mediated condensation reaction between pyridine-3,4-diamine and 2-hydroxy-1-napthaldehyde. The synthesized probe was characterized using 1H and 13C NMR, and HR-Mass spectroscopic techniques. The probe (R) is non-emissive in nature, but on recognition of Al3+, the probe R showed "turn-on" emission (bright yellow colour) showing two emission maxima (522 nm and 547 nm), and no naked eye observable color change. Other competing cations do not show any noticeable fluorescence outcome. The R + Al3+ ensemble can specifically detect L-cysteine among all the essential amino acids by showing a fluorescence "turn-off" response. The sensing mechanism of Al3+ is obeying the chelation-enhanced fluorescence (CHEF) effect. The binding constant of R + Al3+ is 0.3 × 104 M-1. The limit of detection (LoD) for Al3+ and L-cysteine are 2.02 × 10-7 M and 0.5 × 10-5 M respectively. The probe (R) can show maximum efficiency within the pH range (7.0-10.0). The probe is found non-toxic (>80 % cell viability with 15 µM concentration) and employed for the in-vitro fluorescence imaging in the HeLa cell.

Bio-net dataset: AI-based diagnostic solutions using peripheral blood smear images.

Peripheral blood smear examination is one of the basic steps in the evaluation of different blood cells. It is a confirmatory step after an automated complete blood count analysis. Manual microscopy is time-consuming and requires professional laboratory expertise. Therefore, the turn-around time for peripheral smear in a health care center is approximately 3-4 hours. To avoid the traditional method of manual counting under the microscope a computerized automation of peripheral blood smear examination has been adopted, which is a challenging task in medical diagnostics. In recent times, deep learning techniques have overcome the challenges associated with human microscopic evaluation of peripheral smears and this has led to reduced cost and precise diagnosis. However, their application can be significantly improved by the availability of annotated datasets. This study presents a large customized annotated blood cell dataset (named the Bio-Net dataset from healthy individuals) and blood cell detection and counting in the peripheral blood smear images. A mini-version of the dataset for specialized WBC-based image processing tasks is also equipped to classify the healthy and mature WBCs in their respective classes. An object detection algorithm called You Only Look Once (YOLO) with a refashion disposition has been trained on the novel dataset to automatically detect and classify blood cells into RBCs, WBCs, and platelets and compare the results with other publicly available datasets to highlight the versatility. In short the introduction of the Bio-Net dataset and AI-powered detection and counting offers a significant potential for advancement in biomedical research for analyzing and understanding biological data.

A Hydrogen Bonded Non-Porous Organic-Inorganic Framework for Measuring Cysteine in Blood Plasma and Endogenous Cancer Cell.

An imbalance in cysteine (Cys) levels in the cells and plasma has been identified as the risk indicator for various human diseases. The structural similarity of cysteine with its congener homocysteine and glutathione offers challenges in its measurement. Herein, we report a hydrogen-bonded organic-inorganic framework of Cu(II) (HOIF) for the selective detection of cysteine over other biothiols. The non-fluorescent HOIF showed 12-fold green emission in the presence of cysteine. The monomeric unit of HOIF is stabilized via intermolecular hydrogen bonds, resulting in a non-porous network structure. Non-interference from homocysteine, glutathione, and other competitive bio-analytes revealed explicit affinity of HOIF for cysteine. Fluorimetric titration showed a wide working concentration window (650 nM-800 µM) for measuring cysteine in an aqueous medium. The mechanistic investigation involving HRMS, EPR, and UV-vis spectroscopic studies revealed the decomplexation of HOIF with Cys, resulting in a fluorescence turn-on response from the luminescent ligand. Validation using a commercial dye, "Cysteine Green", confirmed the prospect of HOIF for early diagnostic purposes. Utilizing the fluorescence turn-on property of HOIF in the presence of cysteine, we measured cysteine quantitatively in the blood plasma samples. Bio-imaging of endogenous cysteine in cancer cells indicated the ability of HOIF to monitor the intracellular cysteine.

Quantification of cytosolic 'free' calcium in isolated coral cells with confocal microscopy.

Despite its prominent role as an intracellular messenger in all organisms, cytosolic free calcium ([Ca2+]i) has never been quantified in corals or cnidarians in general. Ratiometric calcium dyes and cell imaging have been key methods in successful research on [Ca2+]i in model systems, and could be applied to corals. Here, we developed a procedure to quantify [Ca2+]i in isolated cells from the model coral species Stylophora pistillata using Indo-1 and confocal microscopy. We quantified [Ca2+]i in coral cells with and without intracellular dinoflagellate symbionts, and verified our procedure on cultured mammalian cells. We then used our procedure to measure changes in [Ca2+]i in coral cells exposed to a classic inhibitor of [Ca2+]i regulation, thapsigargin, and also used it to record elevations in [Ca2+]i in coral cells undergoing apoptosis. Our procedure paves the way for future studies into intracellular calcium in corals and other cnidarians.

Fluorescent Carbon Dots: Aggregation-Induced Emission Enhancement, Application as Probe for CN- and Cr2O7-2, Sensing Strips and Bio-imaging Agent.

Fluorescent carbon dots (Trp-CDs) were prepared using tryptophan as precursor and were characterized on the basis of elemental analysis, powder-XRD, IR, Raman spectroscopy, 13C-NMR, UV-Vis, fluorescence and TEM. Trp-CDs exhibit poor fluorescence in 100% water but showed strong Aggregation Induced Emission (AIE) in ethanol and higher alcohols. The anion sensing study of Trp-CD revealed that it selectively detects CN- and Cr2O7-2 and from fluorescence quenching titration study, quenching constant, LOD and range of detection were evaluated. The emission life-time of Trp-CD before and after addition of CN- and Cr2O7-2 were measured, the decay curve before addition of anion was best fitted with a bi-exponential function with life-time of τ1 2.79 ns (10.74%) and τ2 18.93 ns (89.26%). The mechanistic study revealed that for CN-, the fluorescence quenching is due to its interaction with protons attached to surface functional groups and for Cr2O7-2, it is due to inner filter effect (IFE). Sensing strips were prepared by coating Trp-CDs onto various solid surfaces including agarose films and were used for detection of CN- and Cr2O7-. Trp-CD was found to be nontoxic and biocompatible and used as staining agent for Artemia and Bacteria (Bacillus Subtilis, Pseudomonas) and detection of CN- and Cr2O7-.

A highly selective probe for fluorometric sensing of cyanide in an aqueous solution and its application in quantitative determination and living cell imaging.

A simple fluorescent probe (KS4) containing multiple reaction sites (phenolic -OH, imine and C = C bonds) is successfully synthesized and characterized using 1H NMR, 13C NMR, mass and single crystal XRD techniques. KS4 exhibits high selectivity towards CN- over a wide range of common anions in H2O:DMSO (1:1 v/v) leading to an amazing turn-on fluorescence at 505 nm via deprotonation of the phenolic -OH group. The limit of detection (1.3 µM) for CN- was much below the standard (1.9 µM) set by the World Health Organization (WHO). Stoichiometry of the interaction between KS4 and CN- was ascertained as 1:1 by the Job's plot method and the binding constant was determined to be 1.5x104 M-1. Density Functional Theory (DFT) and Time-Dependent Density Functional Theory (TD-DFT) based theoretical insight has been appealed to understand the optical properties of KS4 before and after the addition of CN- ion. The probe shows respectable real-time applicability for qualitative detection of CN- in almond and cassava powder as well as quantification in real water samples with excellent recoveries (98.8 - 99.8%). In addition, KS4 is found to safe towards living HeLa cells and successfully applied to the detection of endogenous cyanide ions in HeLa cells.

Probing the interstitial calcium compartment.

Calcium in interstitial fluids is a crucial ion pool for entry into cells through a plethora of calcium-permeable channels. It is also sensed actively by dedicated receptors. While the mechanisms of global calcium homeostasis and regulation in body fluids appear well understood, more efforts and new technology are needed to elucidate local calcium handling in the small and relatively isolated interstitial spaces between cells. Here we review current methodology for monitoring interstitial calcium and highlight the potential of new approaches for its study. In particular, new generations of high-performance low-affinity genetically encoded calcium indicators could allow imaging of calcium in relatively inaccessible intercellular structures in live tissues and organisms.

Recent progress on small molecular temperature-sensitive fluorescent probes.

Temperature is an important biophysical parameter that is closely related with the metabolic activity in living cells. Therefore, the detection of intracellular temperature changes is crucial for exploring temperature-related biological processes. Fluorescence probe is an ideal tool for observing temperature changes in cells, which has many advantages, such as high sensitivity, good selectivity, and noninvasive, and thus aroused the great interest of researchers. In this paper, we summarize the recent progress of organic small molecule temperature-sensitive fluorescence probes in recent years was reviewed. Particularly, we describe the common response mode to the temperature and the practical applications of the probe in living cells and even animal models. Moreover, an outlook regarding temperature detection in clinical applications is discussed. The temperature-sensitive fluorescent probe is a "black box" to many researchers. This review aims to open a window on the prospect of the noninvasive in vivo detection of temperature which is helpful to deeper understand this rich research area.

Analyte Detection: A Decade of Progress in the Development of Optical/Fluorescent Sensing Probes.

The development of selective and sensitive chemical sensors capable of detecting metal ions, anions, neutral species, explosives and hazardous substances, selectively and sensitively has attracted considerable interest of various research groups. The presence of such analytes within the permissible limits is often beneficial, but the excess amounts may lead to lethal effects to both the environment as well as the living organisms. Owing to the toxicity of the heavy metal ions, toxic anions and nitro-aromatics which are main constituents of explosives, the timely detection of these materials is most desirable to ensure safety and security of the mankind. In this personal account, we present several classes of molecular sensors that were specifically designed in our lab during the past decade for detecting several species in solutions, solid state as well as biological media. Modulation of the optical properties in response to the presence of guest species, led to selective and sensitive detection protocols, and was supported by the theoretical studies wherever possible. We have also extended the application of some of these probes for the on-site detection of analytes by developing the paper strips, glass slides and even the wool and cotton fabrics loaded with probes. One such development represents detection of palladium in human urine and blood samples collected from clinical samples. Additionally, the sensing events in some cases have successfully been reproduced in the live cancer cells. Based on the ease and cost-effective synthesis of the molecular probes, we hope that this account shall provide significant information to researchers in understanding the structure dependent sensing capabilities of the molecular probes.

Spectroscopic and Theoretical Studies on the Selective Detection of Cyanide Ions by a Turn-On Fluorescent Chemo-Dosimeter and its Application in Living Cell Imaging.

A new chemo-dosimeter AK4 containing quinoline fluorophore has rationally been designed, synthesised and characterized using 1H and 13C NMR and mass spectral techniques. The probe senses explicitly CN- ion through a dramatic enhancement in fluorescence over other commonly coexistent anions in H2O:DMSO (9:1 v/v) medium over a broad pH range (4-10). 1H NMR titration revealed the deprotonation followed by nucleophilic addition reaction of CN-, which was supported by 13C NMR and mass spectral examinations. The Job's continuous variation method indicated the formation of a 1:1 adduct between AK4 and CN- with a binding constant of 1.62 × 104 M-1. A limit of detection (LOD) towards CN- of 0.69 µM has been determined, which is much lower than the World Health Organization (WHO) recommended limit of CN- in drinking water (1.9 µM). The changes in the optical properties of AK4 upon reaction with CN- were delineated using Density Functional Theory (DFT) and Time-Dependent Density Functional Theory (TD-DFT) calculations. Moreover, fluorescence microscopic studies established that AK4 could be an effective probe for imaging intracellular CN- in HeLa cells.

Novel & Smart Sensing of Acetate and Hypochlorite Ions With Relay Recognition of Cr3+ Ion With Fluorescence Turn-on: Application in Bio-imaging & Real Sample Analysis.

The receptor-bearing anthraquinone chromophore was synthesized by a simple aldamine condensation reaction, and its anion sensing properties were investigated via colorimetric, UV-vis, photoluminescence, and DFT calculations. The synthesized receptor detects both acetate and hypochlorite ions, where remarkable colorimetric transitions were observed from pink to purple for the acetate ion and pink to blue for the hypochlorite ion. Moreover, in the occurrence of the acetate ion, it shows an admirable answer for the Cr3+ ion, which changes its purple color to pink, while no notable change was observed for other ions. The detection limits of receptors with acetate and hypochlorite are 7.1 × 10-7 M and 9.4 × 10-7 M, respectively. The DFT calculation was performed to better understand the sensing mechanisms of both AcO- and ClO- ions. Furthermore, receptors were effectively utilized in the preparation of optical sensors supported by silica gel for the detection of AcO- and ClO- ions. The receptor proved itself to be potentially useful for real-life application by sensing AcO - in vinegar and ClO - ions in ala. Furthermore, its preeminent detection properties enabled the successful labeling of the AcO- ion in living biological cells.

Quantum Dot Nanomaterials: Preparation, Characterization, Advanced Bio-Imaging and Therapeutic Applications.

The bio-imaging technology is one of the most significant modern applications used in several fields, including early diagnosis of many illnesses that are most important diseases facing humanity and other vital uses. The primary advancement in nanotechnology is the creation of innovative fluorescence probes called quantum dots (QDs). The use of molecular tagging in research, in vivo, and in vitro studies is revolutionized by quantum dots. The application of QD indicates conversion in natural imaging and photography has demonstrated extraordinary appropriateness in bio-imaging, the discovery of novel drugs, and delivery of targeted genes, biosensing, photodynamic therapy, and diagnosis. New potential methods of early cancer detection and treatment management are being researched as a result of the special physical and chemical characteristics of QD probes. The bio-imaging technique depends on the fluorescent emission of the used materials, which is paired with living cells that are easy to see it in 3D without any surgical intervention. Therefore, the use of QDs many types that have unique and appropriate properties for use in that application; In terms of fluorescent emission strength, duration and luminosity.This review article displays some methods of preparation for QDs nanomaterials and the devices used in this. In addition, it presentssome of challenges that must be avoided for the possibility of using them in the bio-imaging field; as toxicity, bio-compatibility, and hydrophilization. It's reviewed some of the devices that use QDs in bio-imaging technique, the QDs application in cell analysis-imaging, and QDs application in vivo imaging.

Tris(2,2'-bipyridyl)ruthenium (II) complex as a universal reagent for the fabrication of heterogeneous electrochemiluminescence platforms and its recent analytical applications.

In recent years, electrochemiluminescence (ECL) has received enormous attention and has emerged as one of the most successful tools in the field of analytical science. Compared with homogeneous ECL, the heterogeneous (or solid-state) ECL has enhanced the rate of the electron transfer kinetics and offers rapid response time, which is highly beneficial in point-of-care and clinical applications. In ECL, the luminophore is the key element, which dictates the overall performance of the ECL-based sensors in various analytical applications. Tris(2,2'-bipyridyl)ruthenium (II) complex, Ru(bpy)32+, is a coordination compound, which is the gold-standard luminophore in ECL. It has played a key role in translating ECL from a "laboratory curiosity" to a commercial analytical instrument for diagnosis. The aim of the present review is to provide the principles of ECL and classical reaction mechanisms-particularly involving the heterogeneous Ru(bpy)32+/co-reactant ECL systems, as well as the fabrication methods and its importance over solution-phase Ru(bpy)32+ ECL. Then, we discussed the emerging technology in solid-state Ru(bpy)32+ ECL-sensing platforms and their recent potential analytical applications such as in immunoassay sensors, DNA sensors, aptasensors, bio-imaging, latent fingerprint detection, point-of-care testing, and detection of non-biomolecules. Finally, we also briefly cover the recent advances in solid-state Ru(bpy)32+ ECL coupled with the hyphenated techniques.

DiceCT for fishes: recommendations for pairing iodine contrast agents with µCT to visualize soft tissues in fishes.

Computed tomography (CT) scanning and other high-throughput three-dimensional (3D) visualization tools are transforming the ways we study morphology, ecology and evolutionary biology research beyond generating vast digital repositories of anatomical data. Contrast-enhanced chemical staining methods, which render soft tissues radio-opaque when coupled with CT scanning, encompass several approaches that are growing in popularity and versatility. Of these, the various diceCT techniques that use an iodine-based solution like Lugol's have provided access to an array of morphological data sets spanning extant vertebrate lineages. This contribution outlines straightforward means for applying diceCT techniques to preserved museum specimens of cartilaginous and bony fishes, collectively representing half of vertebrate species diversity. This study contrasts the benefits of using either aqueous or ethylic Lugol's solutions and reports few differences between these methods with respect to the time required to achieve optimal tissue contrast. It also explores differences in minimum stain duration required for different body sizes and shapes and provides recommendations for staining specimens individually or in small batches. As reported by earlier studies, the authors note a decrease in pH during staining with either aqueous or ethylic Lugol's. Nonetheless, they could not replicate the drastic declines in pH reported elsewhere. They provide recommendations for researchers and collections staff on how to incorporate diceCT into existing curatorial practices, while offsetting risk to specimens. Finally, they outline how diceCT with Lugol's can aid ichthyologists of all kinds in visualizing anatomical structures of interest: from brains and gizzards to gas bladders and pharyngeal jaw muscles.

Nucleic Acid Probes in Bio-Imaging and Diagnostics: Recent Advances in ODN-Based Fluorescent and Surface-Enhanced Raman Scattering Nanoparticle and Nanostructured Systems.

Raman nanoparticle probes are a potent class of optical labels for the interrogation of pathological and physiological processes in cells, bioassays, and tissues. Herein, we review the recent advancements in fluorescent and Raman imaging using oligodeoxyribonucleotide (ODN)-based nanoparticles and nanostructures, which show promise as effective tools for live-cell analysis. These nanodevices can be used to investigate a vast number of biological processes occurring at various levels, starting from those involving organelles, cells, tissues, and whole living organisms. ODN-based fluorescent and Raman probes have contributed to the achievement of significant advancements in the comprehension of the role played by specific analytes in pathological processes and have inaugurated new possibilities for diagnosing health conditions. The technological implications that have emerged from the studies herein described could open new avenues for innovative diagnostics aimed at identifying socially relevant diseases like cancer through the utilization of intracellular markers and/or guide surgical procedures based on fluorescent or Raman imaging. Particularly complex probe structures have been developed within the past five years, creating a versatile toolbox for live-cell analysis, with each tool possessing its own strengths and limitations for specific studies. Analyzing the literature reports in the field, we predict that the development of ODN-based fluorescent and Raman probes will continue in the near future, disclosing novel ideas on their application in therapeutic and diagnostic strategies.

Encapsulation of Dual-Passivated Perovskite Quantum Dots for Bio-Imaging.

Due to their marvelous electrical and optical properties, perovskite nanocrystals have reached remarkable landmarks in solar cells, light-emitting diodes, and photodetectors. However, the intrinsic instability of ionic perovskites, which would undergo an undesirable phase transition and decompose rapidly in ambient humidity, limits their long-term practical deployment. To address this challenge, halogenated trimethoxysilane as the passivation additive is chosen, which utilizes simultaneous halide and silica passivation to enhance the stability of perovskite nanoparticles via a dual-passivation mechanism. The processable nanoparticles show high photoluminescence quantum yield, tunable fluorescence wavelength, and excellent resistance against air and water, highlighting great potential as green to deep-red bio-labels after further phospholipid encapsulation. This work demonstrates that the dual-passivation mechanism could be used to maintain the long-term stability of ionic crystals, which sheds light on the opportunity of halide perovskite nanoparticles for usage in a humid environment.

Supramolecular biomaterials for bio-imaging and imaging-guided therapy.

Benefiting from their unique advantages, including reversibly switchable structures, good biocompatibility, facile functionalization, and sensitive response to biological stimuli, supramolecular biomaterials have been widely applied in biomedicine. In this review, the representative achievements and trends in the design of supramolecular biomaterials (mainly those derived from biomacromolecules) with specific macromolecules including peptides, deoxyribonucleic acid, and polysaccharides, as well as their applications in bio-imaging and imaging-guided therapy are summarized. This review will serve as an important summary and "go for" reference for explorations of the applications of supramolecular biomaterials in bio-imaging and image-guided therapy, and will promote the development of supramolecular chemistry as an emerging interdisciplinary research area.

Emerging Trends in Fluorescence Bioimaging of Divalent Metal Cations Using Small-Molecule Indicators.

Visualization of cation dynamics inside a living system represent a major breakthrough at the crossroad of chemistry and cellular physiology. Since the inception of BAPTA-based cellular calcium indicators in the 1980s, generations of chemical and genetically encoded ion indicators spanning the visible spectrum have been developed. In this article, we bring up three emerging concepts in this field: 1. red-shifting cation indicators towards far-red and near-infrared (NIR) channels; 2. directing the indicators to various subcellular localizations; 3. lowering the phototoxicity of indicators for long term recording. These initiatives collectively echo the advocate of 4D cellular physiology, where biological processes within living systems can be panoramically unveiled under 3D, long-term, and multi-channel imaging with unprecedented spatial and temporal resolution. This outlook poses exciting challenges and opportunities for chemists to upgrade the toolkit of fluorescent indicators as key enablers for a new era of imageomics.

Comparison of Three 3D Segmentation Software Tools for Hip Surgical Planning.

In hip arthroplasty, preoperative planning is fundamental to reaching a successful surgery. Nowadays, several software tools for computed tomography (CT) image processing are available. However, research studies comparing segmentation tools for hip surgery planning for patients affected by osteoarthritic diseases or osteoporotic fractures are still lacking. The present work compares three different software from the geometric, dimensional, and usability perspectives to identify the best three-dimensional (3D) modelling tool for the reconstruction of pathological femoral heads. Syngo.via Frontier (by Siemens Healthcare) is a medical image reading and post-processing software that allows low-skilled operators to produce prototypes. Materialise (by Mimics) is a commercial medical modelling software. 3D Slicer (by slicer.org) is an open-source development platform used in medical and biomedical fields. The 3D models reconstructed starting from the in vivo CT images of the pathological femoral head are compared with the geometries obtained from the laser scan of the in vitro bony specimens. The results show that Mimics and 3D Slicer are better for dimensional and geometric accuracy in the 3D reconstruction, while syngo.via Frontier is the easiest to use in the hospital setting.