Beta Cyclodextrin Conjugated Au-Fe3O4 Janus Nanoparticles with Enhanced Chemo-Photothermal Therapy Performance.

The strategic integration of multi-functionalities within a singular nanoplatform has received growing attention for enhancing treatment efficacy, particularly in chemo-photothermal therapy. This study introduces a comprehensive concept of Janus nanoparticles (JNPs) composed of Au and Fe3O4 nanostructures intricately bonded with ß-cyclodextrins (ß-CD) to encapsulate 5-Fluorouracil (5-FU) and Ibuprofen (IBU). This strategic structure is engineered to exploit the synergistic effects of chemo-photothermal therapy, underscored by their exceptional biocompatibility and photothermal conversion efficiency (∼32.88%). Furthermore, these ß-CD-conjugated JNPs enhance photodynamic therapy by generating singlet oxygen (1O2) species, offering a multi-modality approach to cancer eradication. Computer simulation results were in good agreement with in vitro and in vivo assays. Through these studies, we were able to prove the improved tumor ablation ability of the drug-loaded ß-CD-conjugated JNPs, without inducing adverse effects in tumor-bearing nude mice. The findings underscore a formidable tumor ablation potency of ß-CD-conjugated Au-Fe3O4 JNPs, heralding a new era in achieving nuanced, highly effective, and side-effect-free cancer treatment modalities. STATEMENT OF SIGNIFICANCE: The emergence of multifunctional nanoparticles marks a pivotal stride in cancer therapy research. This investigation unveils Janus nanoparticles (JNPs) amalgamating gold (Au), iron oxide (Fe3O4), and ß-cyclodextrins (ß-CD), encapsulating 5-Fluorouracil (5-FU) and Ibuprofen (IBU) for synergistic chemo-photothermal therapy. Demonstrating both biocompatibility and potent photothermal properties (∼32.88%), these JNPs present a promising avenue for cancer treatment. Noteworthy is their heightened photodynamic efficiency and remarkable tumor ablation capabilities observed in vitro and in vivo, devoid of adverse effects. Furthermore, computational simulations validate their interactions with cancer cells, bolstering their utility as an emerging therapeutic modality. This endeavor pioneers a secure and efficacious strategy for cancer therapy, underscoring the significance of ß-CD-conjugated Au-Fe3O4 JNPs as innovative nanoplatforms with profound implications for the advancement of cancer therapy.

Hyaluronic acid functionalized iron-platinum nanoparticles for photothermal therapy and photoacoustic imaging.

This study represents an innovative approach to construct multi-functional nanoplatforms for cancer diagnosis and therapy by combining hyaluronic acid (HA) with iron-platinum nanoparticles (FePt NPs). These HA-coated FePt NPs, referred to as FePt@HA NPs, demonstrated remarkable biocompatibility, high absorption, and excellent light-to-heat conversion properties in the near-infrared (NIR) region, making them ideal candidates for photothermal therapy (PTT). In vitro studies revealed their effective cancer cell eradication under NIR laser irradiation, while in vivo experiments on mice showcased their superior heating capabilities. Moreover, FePt@HA NPs exhibited a distinct and strong photoacoustic (PA) signal, facilitating enhanced and precise intra-tumoral PA imaging. Our results highlight the potential of FePt@HA NPs as promising photothermal agents for future PTT applications. They offer high selectivity, precision, and minimal side effects in cancer treatment, along with their valuable PA imaging application for tumor localization and characterization.

Hydroxyapatite: A journey from biomaterials to advanced functional materials.

Hydroxyapatite (HAp), a well-known biomaterial, has witnessed a remarkable evolution over the years, transforming from a simple biocompatible substance to an advanced functional material with a wide range of applications. This abstract provides an overview of the significant advancements in the field of HAp and its journey towards becoming a multifunctional material. Initially recognized for its exceptional biocompatibility and bioactivity, HAp gained prominence in the field of bone tissue engineering and dental applications. Its ability to integrate with surrounding tissues, promote cellular adhesion, and facilitate osseointegration made it an ideal candidate for various biomedical implants and coatings. As the understanding of HAp grew, researchers explored its potential beyond traditional biomaterial applications. With advances in material synthesis and engineering, HAp began to exhibit unique properties that extended its utility to other disciplines. Researchers successfully tailored the composition, morphology, and surface characteristics of HAp, leading to enhanced mechanical strength, controlled drug release capabilities, and improved biodegradability. These modifications enabled the utilization of HAp in drug delivery systems, biosensors, tissue engineering scaffolds, and regenerative medicine applications. Moreover, the exceptional biomineralization properties of HAp allowed for the incorporation of functional ions and molecules during synthesis, leading to the development of bioactive coatings and composites with specific therapeutic functionalities. These functionalized HAp materials have demonstrated promising results in antimicrobial coatings, controlled release systems for growth factors and therapeutic agents, and even as catalysts in chemical reactions. In recent years, HAp nanoparticles and nanostructured materials have emerged as a focal point of research due to their unique physicochemical properties and potential for targeted drug delivery, imaging, and theranostic applications. The ability to manipulate the size, shape, and surface chemistry of HAp at the nanoscale has paved the way for innovative approaches in personalized medicine and regenerative therapies. This abstract highlights the exceptional evolution of HAp, from a traditional biomaterial to an advanced functional material. The exploration of novel synthesis methods, surface modifications, and nanoengineering techniques has expanded the horizon of HAp applications, enabling its integration into diverse fields ranging from biomedicine to catalysis. Additionally, this manuscript discusses the emerging prospects of HAp-based materials in photocatalysis, sensing, and energy storage, showcasing its potential as an advanced functional material beyond the realm of biomedical applications. As research in this field progresses, the future holds tremendous potential for HAp-based materials to revolutionize medical treatments and contribute to the advancement of science and technology.

OrganoidChip facilitates hydrogel-free immobilization for fast and blur-free imaging of organoids.

Organoids are three-dimensional structures of self-assembled cell aggregates that mimic anatomical features of in vivo organs and can serve as in vitro miniaturized organ models for drug testing. The most efficient way of studying drug toxicity and efficacy requires high-resolution imaging of a large number of organoids acquired in the least amount of time. Currently missing are suitable platforms capable of fast-paced high-content imaging of organoids. To address this knowledge gap, we present the OrganoidChip, a microfluidic imaging platform that incorporates a unique design to immobilize organoids for endpoint, fast imaging. The chip contains six parallel trapping areas, each having a staging and immobilization chamber, that receives organoids transferred from their native culture plates and anchors them, respectively. We first demonstrate that the OrganoidChip can efficiently immobilize intestinal and cardiac organoids without compromising their viability and functionality. Next, we show the capability of our device in assessing the dose-dependent responses of organoids' viability and spontaneous contraction properties to Doxorubicin treatment and obtaining results that are similar to off-chip experiments. Importantly, the chip enables organoid imaging at speeds that are an order of magnitude faster than conventional imaging platforms and prevents the acquisition of blurry images caused by organoid drifting, swimming, and fast stage movements. Taken together, the OrganoidChip is a promising microfluidic platform that can serve as a building block for a multiwell plate format that can provide high-throughput and high-resolution imaging of organoids in the future.

Yb-Gd Codoped Hydroxyapatite as a Potential Contrast Agent for Tumor-Targeted Biomedical Applications.

Recently, various nanomaterials based on hydroxyapatite (HAp) have been developed for bioimaging applications. In particular, HAp doped with rare-earth elements has attracted significant attention, owing to its enhanced bioactivity and imaging properties. In this study, the wet precipitation method was used to synthesize HAp codoped with Yb and Gd. The synthesized Ybx-Gdx-HAp nanoparticles (NPs) were characterized via various techniques to analyze the crystal phase, functional groups, thermal characteristics, and particularly, the larger surface area. The IR783 fluorescence dye and a folic acid (FA) receptor were conjugated with the synthesized Ybx-Gdx-HAp NPs to develop an effective imaging contrast agent. The developed FA/IR783/Yb-Gd-HAp nanomaterial exhibited improved contrast, sensitivity, and tumor-specific properties, as demonstrated by using the customized LUX 4.0 fluorescence imaging system. An in vitro cytotoxicity study was performed to verify the biocompatibility of the synthesized NPs using MTT assay and fluorescence staining. Photodynamic therapy (PDT) was also applied to determine the photosensitizer properties of the synthesized Ybx-Gdx-HAp NPs. Further, reactive oxygen species generation was confirmed by Prussian blue decay and a 2',7'-dichlorofluorescin diacetate study. Moreover, MDA-MB-231 breast cancer cells were used to evaluate the efficiency of Ybx-Gdx-HAp NP-supported PDT.

Spatial single-cell sequencing of meiosis I arrested oocytes indicates acquisition of maternal transcripts from the soma.

Maternal RNAs are stored from minutes to decades in oocytes throughout meiosis I arrest in a transcriptionally quiescent state. Recent reports, however, propose a role for nascent transcription in arrested oocytes. Whether arrested oocytes launch nascent transcription in response to environmental or hormonal signals while maintaining the meiosis I arrest remains undetermined. We test this by integrating single-cell RNA sequencing, RNA velocity, and RNA fluorescence in situ hybridization on C. elegans meiosis I arrested oocytes. We identify transcripts that increase as the arrested meiosis I oocyte ages, but rule out extracellular signaling through ERK MAPK and nascent transcription as a mechanism for this increase. We report transcript acquisition from neighboring somatic cells as a mechanism of transcript increase during meiosis I arrest. These analyses provide a deeper view at single-cell resolution of the RNA landscape of a meiosis I arrested oocyte and as it prepares for oocyte maturation and fertilization.

Femtosecond laser microdissection for isolation of regenerating C. elegans neurons for single-cell RNA sequencing.

Our understanding of nerve regeneration can be enhanced by delineating its underlying molecular activities at single-neuron resolution in model organisms such as Caenorhabditis elegans. Existing cell isolation techniques cannot isolate neurons with specific regeneration phenotypes from C. elegans. We present femtosecond laser microdissection (fs-LM), a single-cell isolation method that dissects specific cells directly from living tissue by leveraging the micrometer-scale precision of fs-laser ablation. We show that fs-LM facilitates sensitive and specific gene expression profiling by single-cell RNA sequencing (scRNA-seq), while mitigating the stress-related transcriptional artifacts induced by tissue dissociation. scRNA-seq of fs-LM isolated regenerating neurons revealed transcriptional programs that are correlated with either successful or failed regeneration in wild-type and dlk-1 (0) animals, respectively. This method also allowed studying heterogeneity displayed by the same type of neuron and found gene modules with expression patterns correlated with axon regrowth rate. Our results establish fs-LM as a spatially resolved single-cell isolation method for phenotype-to-genotype mapping.

Computational analysis of drug free silver triangular nanoprism theranostic probe plasmonic behavior for in-situ tumor imaging and photothermal therapy.

INTRODUCTION: The advanced features of plasmonic nanomaterials enable initial high accuracy detection with different therapeutic intervention. Computational simulations could estimate the plasmonic heat generation with a high accuracy and could be reliably compared to experimental results. This proposed combined theoretical-experimental strategy may help researchers to better understand other nanoparticles in terms of plasmonic efficiency and usability for future nano-theranostic research. OBJECTIVES: To develop innovative computationally-driven approach to quantify any plasmonic nanoparticles photothermal efficiency and effects before their use as therapeutic agents. METHODS: This report introduces drug free plasmonic silver triangular nanoprisms coated with polyvinyl alcohol biopolymer (PVA-SNT), for in vivo photoacoustic imaging (PAI) guided photothermal treatment (PTT) of triple-negative breast cancer mouse models. The synthesized PVA-SNT nanoparticles were characterized and a computational electrodynamic analysis was performed to evaluate and predict the optical and plasmonic photothermal properties. The in vitro biocompatibility and in vivo tumor abalation study was performed with MDA-MB-231 human breast cancer cell line and in nude mice model. RESULTS: The drug free 140 µg∙mL-1 PVA-SNT nanoparticles with 1.0 W∙cm-2 laser irradiation for 7 min proved to be an effective and optimized theranostic approach in terms of PAI guided triple negative breast cancer treatment. The PVA-SNT nanoparticles exhibits excellent biosafety, photostability, and strong efficiency as PAI contrast agent to visualize tumors. Histological analysis and fluorescence-assisted cell shorter assay results post-treatment apoptotic cells, more importantly, it shows substantial damage to in vivo tumor tissues, killing almost all affected cells, with no recurrence. CONCLUSION: This is a first complete study on computational simulations to estimate the plasmonic heat generation followed by drug free plasmonic PAI guided PTT for cancer treatment. This computationally-driven theranostic approach demonstrates an innovative thought regarding the nanoparticles shape, size, concentration, and composition which could be useful for the prediction of photothermal heat generation in precise nanomedicine applications.

In vivo mimicking injectable self-setting composite bio-cement: Scanning acoustic diagnosis and biological property evaluation for tissue engineering applications.

Injectability and self-setting properties are important factors to increase the efficiency of bone regeneration and reconstruction, thereby reducing the invasiveness of hard tissue engineering procedures. In this study, 63S bioactive glass (BG), nano-hydroxyapatite (n-HAp), alumina, titanium dioxide, and methylene bis-acrylamide (MBAM)-mediated polymeric crosslinking composites were prepared for the formulation of an efficient self-setting bone cement. According to the cytocompatibility and physicochemical analyses, all the samples qualified the standard of the bio-composite materials. They revealed high thermal stability, injectability, and self-setting ability supported by ~ 10.73% (maximum) mass loss, ~ 92-93% injectability and 24 ± 5 min of initial setting time. Moreover, a cellular adhesion and proliferation study was additionally performed with osteoblasts like MG-63 cells, which facilitate pseudopod-like cellular extensions on the BG/n-HAp composite scaffold surface. The SAM study was employed to non-invasively assess the self-setting properties of the composite bio-cement using the post injected distribution and physical properties of the phantom. These results validate the significant potential characteristics of the BG/n-HAp self-setting bio-cement (16:4:2:1) for promising minimal-invasive bone tissue engineering applications.

Design and Micro-Fabrication of Focused High-Frequency Needle Transducers for Medical Imaging.

In this study, we report an advanced fabrication technique to develop a miniature focused needle transducer. Two different types of high-frequency (100 MHz) transducers were fabricated using the lead magnesium niobate-lead titanate (PMN-0.3PT) and lithium niobate (LiNbO3) single crystals. In order to enhance the transducer's performance, a unique mass-spring matching layer technique was adopted, in which gold and parylene play the roles of the mass layer and spring layer, respectively. The PMN-0.3PT transducer had a 103 MHz center frequency with a -6 dB bandwidth of 52%, and a signal-to-noise ratio (SNR) of 42 dB. The center frequency, -6 dB bandwidth, and SNR of the LiNbO3 transducer were 105 MHz, 66%, and 44 dB, respectively. In order to compare and evaluate the transducers' performances, an ultrasonic biomicroscopy (UBM) imaging on the fish eye was performed. The results showed that the LiNbO3 transducer had a better contrast resolution compared to the PMN-0.3PT transducer. The fabricated transducer showed an excellent performance with high-resolution corneal epithelium imaging of the experimental fish eye. These interesting findings are useful for the future biomedical implementation of the fabricated transducers in the field of high-resolution ultrasound imaging and diagnosis purpose.

A Simple Microfluidic Chip for Long-Term Growth and Imaging of Caenorhabditis elegans.

Caenorhabditis elegans (C. elegans) have proved to be a valuable model system for studying developmental and cell biological processes. Understanding these biological processes often requires long-term and repeated imaging of the same animal. Long recovery times associated with conventional immobilization methods done on agar pads have detrimental effects on animal health making it inappropriate to repeatedly image the same animal over long periods of time. This paper describes a microfluidic chip design, fabrication method, on-chip C. elegans culturing protocol, and three examples of long-term imaging to study developmental processes in individual animals. The chip, fabricated with polydimethylsiloxane and bonded on a cover glass, immobilizes animals on a glass substrate using an elastomeric membrane that is deflected using nitrogen gas. Complete immobilization of C. elegans enables robust time-lapse imaging of cellular and sub-cellular events in an anesthetic-free manner. A channel geometry with a large cross-section allows the animal to move freely within two partially sealed isolation membranes permitting growth in the channel with a continuous food supply. Using this simple chip, imaging of developmental phenomena such as neuronal process growth, vulval development, and dendritic arborization in the PVD sensory neurons, as the animal grows inside the channel, can be performed. The long-term growth and imaging chip operates with a single pressure line, no external valves, inexpensive fluidic consumables, and utilizes standard worm handling protocols that can easily be adapted by other laboratories using C. elegans.

Fluorescence conjugated nanostructured cobalt-doped hydroxyapatite platform for imaging-guided drug delivery application.

Multifunctional nanomaterials developed from hydroxyapatite (HAp) with enhanced biological characteristics have recently attracted attention in the biomedical field. The goal of this study is to investigate the potential applications of cobalt-doped HAp (Co-HAp) in the biomedical imaging and therapeutic applications. The co-precipitation approach was used to substitute different molar concentrations of Ca2+ ions with cobalt (Co2+) in HAp structure. The synthesized Co-HAp nanoparticles were studied using various sophisticated techniques to verify the success rate of the doping method. The specific crystal structure, functional groups, size, morphology, photoluminescence property, and thermal stability of the Co-HAp nanoparticles were analyzed based on the characterization results. The computational modelling of doped and undoped HAp reveals the difference in crystal structure parameters. The cytotoxicity study (MTT assay and AO/PI/Hoechst fluorescence staining) reveals the non-toxic characteristics of Co-HAp nanoparticles on MDA-MB-231 breast cancer cell lines. The DOX was loaded onto Co-HAp, showing the maximum drug loading capacity for 2.0 mol% Co-HAp. Drug release was estimated in five different pH environments with various time intervals over 72 h. Furthermore, 2.0 mol% Co-HAp shows excellent fluorescence sensitivity with FITC-conjugated MDA-MB-231 cell lines. These results suggest that cobalt improved the fluorescence intensity of FITC-labeled HAp nanoparticles. This work highlights the promising application of Co-HAp nanoparticles with significant enhanced fluorescence activity for imaging-guided drug delivery system.

Effect of Mindfulness-Based Stress Reduction on Intraocular Pressure in Patients With Ocular Hypertension: A Randomized Control Trial.

PURPOSE: To evaluate the effect of mindfulness-based stress reduction (MBSR) on intraocular pressure (IOP) in patients with ocular hypertension (OHT). DESIGN: Parallel arm, single masked, randomized controlled trial. METHODS: Sixty patients with ocular hypertension and IOP > 21 and < 30 mmHg were recruited at a tertiary eye care centre in India. Thirty patients (group 1) underwent six weeks of one hour daily MBSR sessions, while the other 30 patients (group 2) were waitlisted and kept on follow-up. The primary outcome was change in IOP (ΔIOP) after six weeks of MBSR. Secondary outcomes were effect on serum cortisol level, diurnal variation of IOP, vessel perfusion and vessel density on optical coherence tomography angiography (OCTA), and quality of life (QOL). RESULTS: At six weeks, a significant decrease in IOP was noted in group 1 (23.05 ± 1.17 to 19.15 ± 1.45 mmHg; P = .001) compared with group 2 (22.55 ± 0.98 mmHg to 22.37 ± 1.07 mmHg; P = .107). The ΔIOP was significantly greater in group 1 (3.93 ± 1.47) than group 2 (0.17 ± 0.58; P = .001). The diurnal fluctuation of IOP decreased in group 1 (4.87 ± 1.13 mmHg to 2.73 ± 0.98 mmHg; P = .001) as compared with group 2 (4.50 ± 0.86 mmHg to 4.30 ± 0.83 mmHg; P = .227). Significant improvement in vessel perfusion, vessel density, and flux index was noted on OCTA in group 1 compared with group 2. Group 1 showed a significant decrease (P ≤ .001) in serum cortisol level and an improved QOL (P = .001). CONCLUSION: Mindfulness-based stress reduction was associated with a significant decrease in IOP and serum cortisol, along with an improvement in optic nerve head perfusion and QOL. Mindfulness-based stress reduction can be considered as a potential treatment option in the management of OHT.

Full-view in vivo skin and blood vessels profile segmentation in photoacoustic imaging based on deep learning.

Photoacoustic (PA) microscopy allows imaging of the soft biological tissue based on optical absorption contrast and spatial ultrasound resolution. One of the major applications of PA imaging is its characterization of microvasculature. However, the strong PA signal from skin layer overshadowed the subcutaneous blood vessels leading to indirectly reconstruct the PA images in human study. Addressing the present situation, we examined a deep learning (DL) automatic algorithm to achieve high-resolution and high-contrast segmentation for widening PA imaging applications. In this research, we propose a DL model based on modified U-Net for extracting the relationship features between amplitudes of the generated PA signal from skin and underlying vessels. This study illustrates the broader potential of hybrid complex network as an automatic segmentation tool for the in vivo PA imaging. With DL-infused solution, our result outperforms the previous studies with achieved real-time semantic segmentation on large-size high-resolution PA images.

piRNAQuest V.2: an updated resource for searching through the piRNAome of multiple species.

PIWI interacting RNAs (piRNAs) have emerged as important gene regulators in recent times. Since the release of our first version of piRNAQuest in 2014, lots of novel piRNAs have been annotated in different species other than human, mouse and rat. Such new developments in piRNA research have led us to develop an updated database piRNAQuest V.2. It consists of 92,77,689 piRNA entries for 25 new species of different phylum along with human, mouse and rat. Besides providing primary piRNA features which include their genomic location, with further information on piRNAs overlapping with repeat elements, pseudogenes and syntenic regions, etc., the novel features of this version includes (i) density based cluster prediction, (ii) piRNA expression profile across various healthy and disease systems and (iii) piRNA target prediction. The concept of density-based piRNA cluster identification is robust as it does not consider parametric distribution in its model. The piRNA expression profile for 21 disease systems including cancer have been hosted in addition to 32 tissue specific piRNA expression profile for various species. Further, the piRNA target prediction section includes both predicted and curated piRNA targets within eight disease systems and developmental stages of mouse testis. Further, users can visualize the piRNA-target duplex structure and the ping-pong signature pattern for all the ping-pong piRNA partners in different species. Overall, piRNAQuest V.2 is an updated user-friendly database which will serve as a useful resource to survey, search and retrieve information on piRNAs for multiple species. This freely accessible database is available at http://dibresources.jcbose.ac.in/zhumur/pirnaquest2.

Fluorescence/photoacoustic imaging-guided nanomaterials for highly efficient cancer theragnostic agent.

Imaging modalities combined with a multimodal nanocomposite contrast agent hold great potential for significant contributions in the biomedical field. Among modern imaging techniques, photoacoustic (PA) and fluorescence (FL) imaging gained much attention due to their non-invasive feature and the mutually supportive characteristic in terms of spatial resolution, penetration depth, imaging sensitivity, and speed. In this present study, we synthesized IR783 conjugated chitosan-polypyrrole nanocomposites (IR-CS-PPy NCs) as a theragnostic agent used for FL/PA dual-modal imaging. A customized FL and photoacoustic imaging system was constructed to perform required imaging experiments and create high-contrast images. The proposed nanocomposites were confirmed to have great biosafety, essentially a near-infrared (NIR) absorbance property with enhanced photostability. The in vitro photothermal results indicate the high-efficiency MDA-MB-231 breast cancer cell ablation ability of IR-CS-PPy NCs under 808 nm NIR laser irradiation. The in vivo PTT study revealed the complete destruction of the tumor tissues with IR-CS-PPy NCs without further recurrence. The in vitro and in vivo results suggest that the demonstrated nanocomposites, together with the proposed imaging systems could be an effective theragnostic agent for imaging-guided cancer treatment.

Ultra-widefield photoacoustic microscopy with a dual-channel slider-crank laser-scanning apparatus for in vivo biomedical study.

Photoacoustic microscopy (PAM) is an important imaging tool that can noninvasively visualize the anatomical structure of living animals. However, the limited scanning area restricts traditional PAM systems for scanning a large animal. Here, we firstly report a dual-channel PAM system based on a custom-made slider-crank scanner. This novel scanner allows us to stably capture an ultra-widefield scanning area of 24 mm at a high B-scan speed of 32 Hz while maintaining a high signal-to-noise ratio. Our system's spatial resolution is measured at ∼3.4 µm and ∼37 µm for lateral and axial resolution, respectively. Without any contrast agent, a dragonfly wing, a nude mouse ear, an entire rat ear, and a portion of mouse sagittal are successfully imaged. Furthermore, for hemodynamic monitoring, the mimicking circulating tumor cells using magnetic contrast agent is rapidly captured in vitro. The experimental results demonstrated that our device is a promising tool for biological applications.

Tracking Mitochondrial Density and Positioning along a Growing Neuronal Process in Individual C. elegans Neuron Using a Long-Term Growth and Imaging Microfluidic Device.

The long cellular architecture of neurons requires regulation in part through transport and anchoring events to distribute intracellular organelles. During development, cellular and subcellular events such as organelle additions and their recruitment at specific sites on the growing axons occur over different time scales and often show interanimal variability thus making it difficult to identify specific phenomena in population averages. To measure the variability in subcellular events such as organelle positions, we developed a microfluidic device to feed and immobilize Caenorhabditis elegans for high-resolution imaging over several days. The microfluidic device enabled long-term imaging of individual animals and allowed us to investigate organelle density using mitochondria as a testbed in a growing neuronal process in vivo Subcellular imaging of an individual neuron in multiple animals, over 36 h in our microfluidic device, shows the addition of new mitochondria along the neuronal process and an increase in the accumulation of synaptic vesicles (SVs) at synapses. Long-term imaging of individual C. elegans touch receptor neurons (TRNs) shows that the addition of new mitochondria takes place along the entire neuronal process length at a rate of ∼0.6 mitochondria/h. The threshold for the addition of a new mitochondrion occurs when the average separation between the two preexisting mitochondria exceeds 24 µm. Our assay provides a new opportunity to move beyond simple observations obtained from in vitro assays to allow the discovery of genes that regulate positioning of mitochondria in neurons.

Response of a Zn(II)-based metal-organic coordination polymer towards trivalent metal ions (Al3+, Fe3+ and Cr3+) probed by spectroscopic methods.

A new zinc-based two-dimensional coordination polymer, [Zn(5-AIP)(Ald-4)]·H2O (5-AIP = 5-amino isophthalate, Ald-4 = aldrithiol-4), 1, has been synthesized at room temperature by the layer diffusion technique. Single-crystal X-ray diffraction analysis of 1 showed a two-dimensional bilayer structure. An aqueous suspension of 1 upon excitation at 300 nm displayed an intense blue emission at 403 nm. The luminescence spectra were interestingly responsive and selective to Al3+, Cr3+ and Fe3+ ions even in the presence of other interfering ions. The calculated detection limits for Al3+, Cr3+ and Fe3+ were 0.35 µM ([triple bond, length as m-dash]8.43 ppb), 0.46 µM ([triple bond, length as m-dash]22.6 ppb) and 0.30 µM ([triple bond, length as m-dash]15.85 ppb), respectively. Notably, with the cumulative addition of Al3+ ions, the luminescence intensity at 403 nm decreased steadily with a gradual red shift up to 427 nm. Afterward, this red shifted peak showed a turn-on effect upon further addition of Al3+ ions. On the other hand, for Cr3+ and Fe3+ ions, there was only drastic luminescence quenching and a large red shift up to 434 nm. This indicated the formation of a complex between 1 and these metal ions, which was also supported by the UV-Visible absorption spectra of 1 that showed the appearance of a new band at 280 nm in the presence of these three metal ions. The FTIR spectra revealed that these ions interacted with the carboxylate oxygen atom of 5-AIP and the nitrogen atom of the Ald-4 ligand in the structure. The luminescence lifetime decay analysis manifested that a charge-transfer type complex was formed between 1 and Cr3+ and Fe3+ ions that resulted in huge luminescence quenching due to the efficient charge transfer involving the vacant d-orbitals, whereas for Al3+ ions having no vacant d-orbital, turn-on of luminescence occurred because of the increased rigidity of 1 upon complexation.