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Cancer, ranked as the second leading cause of global mortality with a prevalence of 1 in 6 deaths, necessitates innovative approaches for effective treatment. Combinatorial drug therapy for cancer treatment targets several key pathways simultaneously and potentially enhances anti-cancer efficacy without intolerable side effects. However, it demands precise and accurate control of drug-dose combinations and their release. In this study, we demonstrated a selectively cross-linked hydrogel-based platform that can quantify and release drugs simultaneously for in-parallel cocktail drug screening. PDMS was used as the flow channel substrate and the poly (ethylene glycol) diacrylate (PEGDA) hydrogel array was formed by UV exposure using the photomask. Employing our platform, cocktails of anticancer drugs are precisely loaded and simultaneously released in-parallel into HCT-116 colon cancer cells, facilitating combinatorial drug screening. The integration of an artificial intelligence-based complex system response (AI-CSR) platform successfully identifies optimal drug-dose combinations from a pool of ten approved drugs. Notably, our cocktail drug chip demonstrates exceptional efficiency, screening 155 drug-dose combinations within a brief two and a half hours, a marked improvement over traditional methods. Furthermore, the device exhibits low drug consumption, requiring a mere 1 μL per patch of chip. Thus, our developed PDMS drug-loaded hydrogel platform presents a novel and expedited approach to quantifying drug concentrations, promising to be a faster, efficient and more precise approach for conducting cocktail drug screening experiments.

Atomic Ag cluster bonding is employed to reinforce the interface between PF3T nano-cluster and TiO 2 nanoparticle. With an optimized Ag loading (Ag/TiO 2  = 0.5 wt%), the Ag atoms will uniformly disperse on TiO 2 thus generating a high density of intermediate states in the band gap to form the electron channel between the terthiophene group of PF3T and the TiO 2 in the hybrid composite (denoted as T@Ag05-P). The former expands the photon absorption band width and the latter facilitates the core-hole splitting by injecting the photon excited electron (from the excitons in PF3T) into the conduction band (CB) of TiO 2 . These characteristics enable the high efficiency of H 2 production to 16 580 µmol h -1  g -1 and photocatalysis stability without degradation under visible light exposure for 96 h. Compared to that of hybrid material without Ag bonding (TiO 2 @PF3T), the H 2 production yield and stability are improved by 4.1 and 18.2-fold which shows the best performance among existing materials in similar component combination and interfacial reinforcement. The unique bonding method offers a new prospect to accelerate the development of photocatalytic hydrogen production technologies., (© 2024 Wiley‐VCH GmbH.)

Background: Most breast cancer-related deaths are caused by distant metastases and drug resistance. It is important to find appropriate biomarkers to monitor the disease and to predict patient responses after treatment early and accurately. Many studies have found that clustered circulating tumor cells, with more correlations with metastatic cancer and poor survival of patients than individual ones, are promising biomarkers., Methods: Eighty samples from eleven patients with breast cancer during follow-up visits were examined. By using a microfluidic chip and imaging system, the number of circulating tumor cells and microemboli (CTC/CTM) were counted to assess the distribution in stratified patients and the potential in predicting the disease condition of patients after treatments during follow-up visits. Specific components and subtypes of CTM were also preliminarily investigated., Results: Compared to CTC, CTM displayed a distinguishable distribution in stratified patients, having a better AUC value, in predicting the disease progression of breast cancer patients during follow-up visits in this study. Four subtypes were categorized from the identified CTM by considering different components. In combination with CEA and CA153, enumerated CTC and CTM from individual patients were applied to monitor the disease condition and patient response to the therapy during follow-up visits., Conclusions: The CTM and its subtypes are promising biomarkers and valuable tools for studying cancer metastasis and longitudinally monitoring cancer patients during follow-up visits.

Background: Listeriosis is caused by the facultative anaerobic bacterium Listeria monocytogenes . Infection from Listeria -contaminated food or water is the main etiology. If Listeria travels outside the intestines, it can cause invasive listeriosis, such as sepsis, meningitis, and meningoencephalitis. Invasive illness is especially dangerous for pregnant women and their newborns, elderly people, and people with compromised immune systems or medical conditions such as end-stage kidney disease (ESKD) patients receiving long-term dialysis., Purpose: Describe the manifestations and hospital outcomes of invasive listeriosis and identify the risk factors for in-hospital and one-year mortality in ESKD patients receiving long-term dialysis., Patients and Methods: This retrospective observational study examined hospitalized patient records at a Taiwanese tertiary medical center from August 1, 2000, to August 31, 2021. ESKD patients on chronic dialysis were identified with invasive listeriosis by blood culture and discharge diagnosis. Over 21 years, we accurately recorded 26 cases., Results: ESKD patients on chronic dialysis with invasive listeriosis have a poor prognosis. Only 53.8% of chronic dialysis patients with invasive listeriosis survived their first hospital episode. 42.3% of hospitalized ESKD patients with invasive listeriosis survived one year later. In univariate analysis, shock, tachypnea (RR ≥ 22), respiratory failure, qSOFA score ≥ 2, and lower initial platelet count were linked to greater in-hospital mortality rates., Conclusion: ESKD patients with invasive listeriosis have a grave prognosis. Our research reveals that an early blood sample for a bacterial culture may identify invasive listeriosis in chronic dialysis patients with fever, nausea or vomiting, confusion, and respiratory distress. This study is the first to identify a lower platelet count and qSOFA score ≥ 2 as markers of high-risk invasive listeriosis in ESKD patients., Competing Interests: The authors report no conflicts of interest in this work., (© 2024 Liu et al.)

Unraveling the intricacies between oxygen dynamics and cellular processes in the tumor microenvironment (TME) hinges upon precise monitoring of intracellular and intratumoral oxygen levels, which holds paramount significance. The majority of these reported oxygen nanoprobes suffer compromised lifetime and quantum yield when exposed to the robust ROS activities prevalent in TME, limiting their prolonged in vitro usability. Herein, the ruthenium-embedded oxygen nano polymeric sensor (Ru-ONPS) is proposed for precise oxygen gradient monitoring within the cellular environment and TME. Ru-ONPS (≈64±7 nm) incorporates [Ru(dpp) 3 ]Cl 2 dye into F-127 and crosslinks it with urea and paraformaldehyde, ensuring a prolonged lifetime (5.4 µs), high quantum yield (66.65 ± 2.43% in N 2 and 49.80 ± 3.14% in O 2 ), superior photostability (>30 min), and excellent stability in diverse environmental conditions. Based on the Stern-Volmer plot, the Ru-ONPS shows complete linearity for a wide dynamic range (0-23 mg L -1 ), with a detection limit of 10 µg mL -1 . Confocal imaging reveals Ru-ONPS cellular uptake and intratumoral distribution. After 72 h, HCT-8 cells show 5.20±1.03% oxygen levels, while NIH3T3 cells have 7.07±1.90%. Co-culture spheroids display declining oxygen levels of 17.90±0.88%, 10.90±0.88%, and 5.10±1.18%, at 48, 120, and 216 h, respectively. Ru-ONPS advances cellular oxygen measurement and facilitates hypoxia-dependent metastatic research and therapeutic target identification., (© 2023 Wiley‐VCH GmbH.)

This study explores the potential of laser-induced nano-photon-poration as a non-invasive technique for the intracellular delivery of micro/macromolecules at the single-cell level. This research proposes the utilization of gold-coated spiky polymeric nanoparticles (Au-PNPs) and gold nanorods (GNRs) to achieve efficient intracellular micro/macromolecule delivery at the single-cell level. By shifting the operating wavelength towards the near-infrared (NIR) range, the intracellular delivery efficiency and viability of Au-PNP-mediated photon-poration are compared to those using GNR-mediated intracellular delivery. Employing Au-PNPs as mediators in conjunction with nanosecond-pulsed lasers, a highly efficient intracellular delivery, while preserving high cell viability, is demonstrated. Laser pulses directed at Au-PNPs generate over a hundred hot spots per particle through plasmon resonance, facilitating the formation of photothermal vapor nanobubbles (PVNBs). These PVNBs create transient pores, enabling the gentle transfer of cargo from the extracellular to the intracellular milieu, without inducing deleterious effects in the cells. The optimization of wavelengths in the NIR region, coupled with low laser fluence (27 mJ/cm 2 ) and nanoparticle concentrations (34 µg/mL), achieves outstanding delivery efficiencies (96%) and maintains high cell viability (up to 99%) across the various cell types, including cancer and neuronal cells. Importantly, sustained high cell viability (90-95%) is observed even 48 h post laser exposure. This innovative development holds considerable promise for diverse applications, encompassing drug delivery, gene therapy, and regenerative medicine. This study underscores the efficiency and versatility of the proposed technique, positioning it as a valuable tool for advancing intracellular delivery strategies in biomedical applications.

Early prognosis of cancer recurrence remains difficult partially due to insufficient and ineffective screening biomarkers or regimes. This study evaluated the rare circulating tumor microemboli (CTM) from liquid biopsy individually and together with circulating tumor cells (CTCs) and serum CEA/CA19-9 in a panel, on early prediction of colorectal cancer (CRC) recurrence. Stained CTCs/CTM were detected by a microfluidic chip-based automatic rare-cell imaging platform. ROC, AUC, Kaplan-Meier survival, and Cox proportional hazard models regarding 4 selected biomarkers were analyzed. The relative risk, odds ratio, predictive accuracy, and positive/negative predictive value of biomarkers individually and in combination, to predict CRC recurrence were assessed and preliminarily validated. The EpCAM + Hochest + CD45 - CTCs/CTM could be found in all cancer stages, where more recurrences were observed in late-stage cases. Significant correlations between CTCs/CTM with metastatic stages and clinical treatment were illustrated. CA19-9 and CTM could be seen as independent risk factors in patient survivals, while stratified patients by grouped biomarkers on the Kaplan-Meier analyses presented more significant differences in predicting CRC recurrences. By monitoring the panel of selected biomarkers, disease progressions of 4 CRC patients during follow-up visits after first treatments within 3 years were predicted successfully. This study unveiled the value of rare CTM on clinical studies and a panel of selected biomarkers on predicting CRC recurrences in patients at the early time after medical treatment, in which the CTM and serum CA19-9 could be applied in clinical surveillance and CRC management to improve the accuracy., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Cell patterning is a powerful technique for the precise control and arrangement of cells, enabling detailed single-cell analysis with broad applications in therapeutics, diagnostics, and regenerative medicine. This study presents a novel and efficient technique that enables massively parallel high throughput cell patterning and precise delivery of small to large biomolecules into patterned cells. The innovative cell patterning device proposed in this study is a standalone, ultrathin 3D SU-8 micro-stencil membrane, with a thickness of 10 μm. It features an array of micro-holes ranging from 40 μm to 80 μm, spaced apart by 50 μm to 150 μm. By culturing cells on top of this SU-8 membrane, the technique achieves highly efficient cell patterns varying from single-cell to cell clusters on a Petri dish. Utilizing this technique, we have achieved a remarkable reproducible patterning efficiency for mouse fibroblast L929 (80.5%), human cervical SiHa (81%), and human neuroblastoma IMR32 (89.6%) with less than 1% defects in undesired areas. Single-cell patterning efficiency was observed to be highest at 75.8% for L929 cells. Additionally, we have demonstrated massively parallel high throughput uniform transfection of large biomolecules into live patterned cells by employing an array of titanium micro-rings (10 μm outer diameter, 3 μm inner diameter) activated through infrared light pulses. Successful delivery of a wide range of small to very large biomolecules, including propidium iodide (PI) dye (668.4 Da), dextran (3 kDa), siRNA (13.3 kDa), and β-galactosidase enzyme (465 kDa), was accomplished in cell patterns for various cancer cells. Notably, our platform achieved exceptional delivery efficiencies of 97% for small molecules like PI dye and 84% for the enzyme, with corresponding high cell viability of 100% and 90%, respectively. Furthermore, the compact and reusable SU-8-based membrane device facilitates highly efficient cell patterning, transfection, and cell viability, making it a promising tool for diagnostics and therapeutic applications.

Rapid and sensitive diagnostics of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is of utmost importance to control the widespread coronavirus disease 2019 (COVID-19) upsurge. This study demonstrated a novel one-pot surface-enhanced Raman scattering (SERS) based immunoassay to detect SARS-CoV-2, without any washing process using a portable Raman spectrometer. The SERS-immune assay was designed using a regular digital versatile disk (DVD) substrate integrated with Raman reporter labeled silver nanoparticles for double clamping effects. The disks were molded to form nanopillar arrays and coated with silver film to enhance the sensitivity of immunoassay. The SERS platform demonstrated a limit of detection (LoD) up to 50 pg mL -1 for SARS-CoV-2 spike protein and virus-like-particle (VLP) protein in phosphate buffer saline within a turnaround time of 20 mins. Moreover, VLP protein spiked in untreated saliva achieved an LoD of 400 pg mL -1 , providing a cycle threshold (Ct) value range of 30-32, closer to reverse transcription-polymerase chain reaction (RT-PCR) results (35-40) and higher than the commercial rapid antigen tests, ranging from 25 to 28. Therefore, the developed one-pot SERS based biosensor exhibited highly sensitive and rapid detection of SARS-CoV-2, which could be a potential point-of-care platform for early and cost-effective diagnosis of the COVID-19 virus., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Elsevier B.V. All rights reserved.)

A hybrid composite of organic-inorganic semiconductor nanomaterials with atomic Au clusters at the interface decoration (denoted as PF3T@Au-TiO 2 ) is developed for visible-light-driven H 2 production via direct water splitting. With a strong electron coupling between the terthiophene groups, Au atoms and the oxygen atoms at the heterogeneous interface, significant electron injection from the PF3T to TiO 2 occurs leading to a quantum leap in the H 2 production yield (18 578 µmol g -1 h -1 ) by ≈39% as compared to that of the composite without Au decoration (PF3T@TiO 2 , 11 321 µmol g -1 h -1 ). Compared to the pure PF3T, such a result is 43-fold improved and is the best performance among all the existing hybrid materials in similar configurations. With robust process control via industrially applicable methods, it is anticipated that the findings and proposed methodologies can accelerate the development of high-performance eco-friendly photocatalytic hydrogen production technologies., (© 2023 Wiley-VCH GmbH.)

This paper proposes a Swiss-roll-type mini-reformer employing a copper-zinc catalyst for high-efficient SRM process. Although the commercially available copper-zinc catalysts commonly used in cylindrical-type reformers provide decent conversion rates in the short term, their long-term durability still requires improvement, mainly due to temperature variations in the reformer, catalyst loading, and thermal sintering issues. This Swiss-roll-shaped mini-reformer is designed to improve thermal energy preservation/temperature uniformity by using dual spiral channels to improve the long-term durability while maintaining methanol-reforming efficiency. It was fabricated on a copper plate that was 80 mm wide, 80 mm long, and 4 mm high with spiral channels that were 2 mm deep, 4 mm wide, and 350 mm long. To optimize the design and reformer operation, the catalyst porosity, gas hourly speed velocity (GHSV), operation temperature, and fuel feeding rate are investigated. Swiss-roll-type reformers may require higher driving pressures but can provide better thermal energy preservation and temperature uniformity, posing a higher conversion rate for the same amount of catalyst when compared with other geometries. By carefully adjusting the catalyst bed porosity, locations, and catalyst loading amount as well as other conditions, an optimized gas hourly space velocity (GHSV) can be obtained (14,580 mL/g·h) and lead to not only a high conversion rate (96%) and low carbon monoxide generation rate (0.98%) but also a better long-term durability (decay from 96% to 88.12% after 60 h operation time) for SRM processes. The decay rate, 0.13%/h, after 60 h of operation, is five-folds lower than that (0.67%/h, 0.134%/h) of a commercial cylindrical-type fixed-bed reactor with a commercial catalyst.

Herein, this work aims to directly visualize the morphological evolution of the controlled self-assembly of star-block polystyrene- block -polydimethylsiloxane (PS- b -PDMS) thin films via in situ transmission electron microscopy (TEM) observations. With an environmental chip, possessing a built-in metal wire-based microheater fabricated by the microelectromechanical system (MEMS) technique, in situ TEM observations can be conducted under low-dose conditions to investigate the development of film-spanning perpendicular cylinders in the block copolymer (BCP) thin films via a self-alignment process. Owing to the free-standing condition, a symmetric condition of the BCP thin films can be formed for thermal annealing under vacuum with neutral air surface, whereas an asymmetric condition can be formed by an air plasma treatment on one side of the thin film that creates an end-capped neutral layer. A systematic comparison of the time-resolved self-alignment process in the symmetric and asymmetric conditions can be carried out, giving comprehensive insights for the self-alignment process via the nucleation and growth mechanism.

Temperature governs the reactivity of a wide range of biomolecules in the cellular environment dynamically. The complex cellular pathways and molecules in solid tumors substantially produce temperature gradients in the tumor microenvironment (TME). Hence, visualization of these temperature gradients at the cellular level would give physiologically relevant spatio-temporal information about solid tumors. This study used fluorescent polymeric nano-thermometers (FPNTs) to measure the intratumor temperature in co-cultured 3D tumor spheroids. A temperature-sensitive rhodamine-B dye and Pluronic F-127 were conjugated through hydrophobic and hydrophobic interactions and then cross-linked with urea-paraformaldehyde resins to form the FPNTs. The characterization results exhibit monodisperse nanoparticles (166 ± 10 nm) with persistent fluorescence. The FPNTs exhibit a linear response with a wide temperature sensing range (25-100 °C) and are stable toward pH, ionic strength, and oxidative stress. FPNTs were utilized to monitor the temperature gradient in co-cultured 3D tumor spheroids and the temperature difference between the core (34.9 °C) and the periphery (37.8 °C) was 2.9 °C. This investigation demonstrates that the FPNTs have great stability, biocompatibility, and high intensity in a biological medium. The usage of FPNTs as a multifunctional adjuvant may demonstrate the dynamics of the TME and they may be suitable candidates to examine thermoregulation in tumor spheroids.

Background: Boron neutron capture therapy (BNCT) is a promising cancer treatment that eliminates tumor cells by triggering high-energy radiation within cancer cells. Aim: In vivo evaluation of poly(vinyl alcohol)/boric acid crosslinked nanoparticles (PVA/BA NPs) for BNCT. Materials & methods: PVA/BA NPs were synthesized and intravenously injected into tumor-bearing mice for BNCT. Results: The in vitro boron uptake of PVA/BA NPs in tumor cells was 70-fold higher than the required boron uptake for successful BNCT. In an in vivo study, PVA/BA NPs showed a 44.29% reduction in tumor size compared with clinically used boronophenylalanine for oral cancer in a murine model. Conclusion: PVA/BA NPs exhibited effective therapeutic results for oral cancer treatments in BNCT.

Personalised drug delivery systems with the ability to offer real-time imaging and control release are an advancement in diagnostic and therapeutic applications. This allows for a tailored drug dosage specific to the patient with a release profile that offers the optimum therapeutic effect. Coupling this application with medical imaging capabilities, real-time contrast can be viewed to display the interaction with the host. Current approaches towards such novelty produce a drug burst release profile and contrasting agents associated with side effects as a result of poor encapsulation of these components. In this study, a 3D-printed drug delivery matrix with real-time imaging is engineered. Polycaprolactone (PCL) forms the bulk structure and encapsulates tetracycline hydrochloride (TH), an antibiotic drug and Iron Oxide Nanoparticles (IONP, Fe 3 O 4 ), a superparamagnetic contrasting agent. Hot melt extrusion (HME) coupled with fused deposition modelling (FDM) is utilised to promote the encapsulation of TH and IONP. The effect of additives on the formation of micropores (10-20 µm) on the 3D-printed surface was investigated. The high-resolution process demonstrated successful encapsulation of both bioactive and nano components to present promising applications in drug delivery systems, medical imaging and targeted therapy.

We previously developed chicken interleukin-1β (IL-1β) mutants as single-dose adjuvants that induce protective immunity when co-administered with an avian vaccine. However, livestock such as pigs may require a vaccine adjuvant delivery system that provides long-lasting protection to reduce the need for successive booster doses. Therefore, we developed chitosan-coated alginate microparticles as a carrier for bovine serum albumin (BSA) or porcine IL-1β (pIL-1β) and assessed their physical, chemical, and biological properties. Electrospraying of the BSA-loaded alginate microparticles (BSA/ALG MPs) resulted in an encapsulation efficiency of 50%, and those MPs were then coated with chitosan (BSA/ALG/CHI MPs). Optical and scanning electron microscopy, zeta potential analysis, and Fourier transform infrared spectroscopy were used to characterize these MPs. The BSA encapsulation parameters were applied to ALG/CHI MPs loaded with pIL-1β, which were not cytotoxic to porcine fibroblasts but had enhanced bio-activity over unencapsulated pIL-1β. The chitosan layer of the BSA/ALG/CHI MPs prevented burst release and facilitated sustained release of pIL-1β for at least 28 days. In conclusion, BSA/ALG/CHI MPs prepared as a carrier for pIL-1β may be used as an adjuvant for the formulation of pig vaccines.

This work aims to demonstrate a facile method for the controlled orientation of nanostructures of block copolymer (BCP) thin films. A simple diblock copolymer system, polystyrene- block -polydimethylsiloxane (PS- b -PDMS), is chosen to demonstrate vacuum-driven orientation for solving the notorious low-surface-energy problem of silicon-based BCP nanopatterning. By taking advantage of the pressure dependence of the surface tension of polymeric materials, a neutral air surface for the PS- b -PDMS thin film can be formed under a high vacuum degree (∼10 -4 Pa), allowing the formation of the film-spanning perpendicular cylinders and lamellae upon thermal annealing. In contrast to perpendicular lamellae, a long-range lateral order for forming perpendicular cylinders can be efficiently achieved through the self-alignment mechanism for induced ordering from the top and bottom of the free-standing thin film.

Insufficient prognosis of local recurrence contributes to the poor progression-free survival rate and death in colorectal cancer (CRC) patients. Various biomarkers have been explored in predicting CRC recurrence. This study investigated the expressions of plasma/exosomal microRNA-21 (miR-21) in 113 CRC patients by qPCR, their values of predicting CRC recurrence, and the possibility to improve the prognostic efficacy in early CRC recurrence in stratified patients by combined biomarkers including circulating miR-21s, circulating tumour cells/microemboli (CTCs/CTM), and serum carcinoembryonic antigen (CEA)/carbohydrate antigen 19-9 (CA19-9). Expressions of plasma and exosomal miR-21s were significantly correlated (p < 0.0001) in all and late-stage patients, presenting similar correlations with other biomarkers. However, stage IV patients stratified by a high level of exosomal miR-21 and stage I to III patients stratified by a high level of plasma miR-21 displayed significantly worse survival outcomes in predicting CRC recurrence, suggesting their different values to predict CRC recurrence in stratified patients. Comparable and even better performances in predicting CRC recurrence in late-stage patients were found by CTCs/CTM from our blood samples as sensitive biomarkers. Improved prognosing efficacy in CRC recurrence and better outcomes to significantly differentiate recurrence in stratified patients could be obtained by analysing combined biomarkers.

Single-neuron actions are the basis of brain function, as clinical sequelae, neuronal dysfunction or failure for most of the central nervous system (CNS) diseases and injuries can be identified via tracing single-neurons. The bulk analysis methods tend to miscue critical information by assessing the population-averaged outcomes. However, its primary requisite in neuroscience to analyze single-neurons and to understand dynamic interplay of neurons and their environment. Microfluidic systems enable precise control over nano-to femto-liter volumes via adjusting device geometry, surface characteristics, and flow-dynamics, thus facilitating a well-defined micro-environment with spatio-temporal control for single-neuron analysis. The microfluidic platform not only offers a comprehensive landscape to study brain cell diversity at the level of transcriptome, genome, and/or epigenome of individual cells but also has a substantial role in deciphering complex dynamics of brain development and brain-related disorders. In this review, we highlight recent advances of microfluidic devices for single-neuron analysis, i.e., single-neuron trapping, single-neuron dynamics, single-neuron proteomics, single-neuron transcriptomics, drug delivery at the single-neuron level, single axon guidance, and single-neuron differentiation. Moreover, we also emphasize limitations and future challenges of single-neuron analysis by focusing on key performances of throughput and multiparametric activity analysis on microfluidic platforms., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2022 Published by Elsevier Ltd.)

This study demonstrates a novel multi-functional microfluidic system, designated three dimensional Alternative Current Electrokinetic/Surface Enhanced Raman Scattering (3D-ACEK/SERS), which can concentrate bacteria from whole blood, identify bacterial species, and determine antibiotic susceptibilities of the bacteria rapidly. The system consists of a hybrid electrokinetic mechanism, integrating AC-electroosmosis (AC-EO) and dielectrophoresis (DEP) that allows thousand-fold concentration of bacteria, including S. aureus, Escherichia coli, and Chryseobacterium indologenes, in the center of an electrode with a wide range of working distance (hundreds to thousands of μm), while exclusion of blood cells through negative DEP forces. This microchip employs SERS assay to determine the identity of the concentrated bacteria in approximately 2 min with a limit of detection of 3 CFU/ml, 5 orders of magnitude lower than that using standard centrifugation-purification process. Finally, label-free antibiotic susceptibility testing has been successfully demonstrated on the platform using both antibiotic-sensitive and multidrug-resistant bacterial strains illustrating a potential utility of the system to clinical applications., (Copyright © 2021. Published by Elsevier B.V.)

Microfluidic platforms gain popularity in biomedical research due to their attractive inherent features, especially in nanomaterials synthesis. This review critically evaluates the current state of the controlled synthesis of nanomaterials using microfluidic devices. We describe nanomaterials' screening in microfluidics, which is very relevant for automating the synthesis process for biomedical applications. We discuss the latest microfluidics trends to achieve noble metal, silica, biopolymer, quantum dots, iron oxide, carbon-based, rare-earth-based, and other nanomaterials with a specific size, composition, surface modification, and morphology required for particular biomedical application. Screening nanomaterials has become an essential tool to synthesize desired nanomaterials using more automated processes with high speed and repeatability, which can't be neglected in today's microfluidic technology. Moreover, we emphasize biomedical applications of nanomaterials, including imaging, targeting, therapy, and sensing. Before clinical use, nanomaterials have to be evaluated under physiological conditions, which is possible in the microfluidic system as it stimulates chemical gradients, fluid flows, and the ability to control microenvironment and partitioning multi-organs. In this review, we emphasize the clinical evaluation of nanomaterials using microfluidics which was not covered by any other reviews. In the future, the growth of new materials or modification in existing materials using microfluidics platforms and applications in a diversity of biomedical fields by utilizing all the features of microfluidic technology is expected., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Highly efficient intracellular delivery strategies are essential for developing therapeutic, diagnostic, biological, and various biomedical applications. The recent advancement of micro/nanotechnology has focused numerous researches towards developing microfluidic device-based strategies due to the associated high throughput delivery, cost-effectiveness, robustness, and biocompatible nature. The delivery strategies can be carrier-mediated or membrane disruption-based, where membrane disruption methods find popularity due to reduced toxicity, enhanced delivery efficiency, and cell viability. Among all of the membrane disruption techniques, the mechanoporation strategies are advantageous because of no external energy source required for membrane deformation, thereby achieving high delivery efficiencies and increased cell viability into different cell types with negligible toxicity. The past two decades have consequently seen a tremendous boost in mechanoporation-based research for intracellular delivery and cellular analysis. This article provides a brief review of the most recent developments on microfluidic-based mechanoporation strategies such as microinjection, nanoneedle arrays, cell-squeezing, and hydroporation techniques with their working principle, device fabrication, cellular delivery, and analysis. Moreover, a brief discussion of the different mechanoporation strategies integrated with other delivery methods has also been provided. Finally, the advantages, limitations, and future prospects of this technique are discussed compared to other intracellular delivery techniques., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Tuhin Subhra Santra reports financial support was provided by the DBT/Wellcome Trust India Alliance Fellowship., (© 2021 Published by Elsevier Ltd.)

We report a one-pot facile synthesis of highly photoresponsive bovine serum albumin (BSA) templated bismuth-copper sulfide nanocomposites (BSA-BiZ/Cu x S NCs, where BiZ represents in situ formed Bi 2 S 3 and bismuth oxysulfides (BOS)). As-formed surface vacancies and BiZ/Cu x S heterojunctions impart superior catalytic, photodynamic and photothermal properties. Upon near-infrared (NIR) irradiation, the BSA-BiZ/Cu x S NCs exhibit broad-spectrum antibacterial activity, not only against standard multidrug-resistant (MDR) bacterial strains but also against clinically isolated MDR bacteria and their associated biofilms. The minimum inhibitory concentration of BSA-BiZ/Cu x S NCs is 14-fold lower than that of BSA-Cu x S NCs because their multiple heterojunctions and vacancies facilitated an amplified phototherapeutic response. As-prepared BSA-BiZ/Cu x S NCs exhibited substantial biofilm inhibition (90%) and eradication (>75%) efficiency under NIR irradiation. Furthermore, MRSA-infected diabetic mice were immensely treated with BSA-BiZ/Cu x S NCs coupled with NIR irradiation by destroying the mature biofilm on the wound site, which accelerated the wound healing process via collagen synthesis and epithelialization. We demonstrate that BSA-BiZ/Cu x S NCs with superior antimicrobial activity and high biocompatibility hold great potential as an effective photosensitive agent for the treatment of biofilm-associated infections.

Three-dimensional (3D) spheroid culture provides opportunities to model tumor growth closer to its natural context. The collagen network in the extracellular matrix supports autonomic tumor cell proliferation, but its presence and role in tumor spheroids remain unclear. In this research, we developed an in vitro 3D co-culture model in a microwell 3D (μ-well 3D) cell-culture array platform to mimic the tumor microenvironment (TME). The modular setup is used to characterize the paracrine signaling molecules and the role of the intraspheroidal collagen network in cancer drug resistance. The μ-well 3D platform is made up of poly(dimethylsiloxane) that contains 630 round wells for individual spheroid growth. Inside each well, the growth surface measured 500 μm in diameter and was functionalized with the amphiphilic copolymer. HCT-8 colon cancer cells and/or NIH3T3 fibroblasts were seeded in each well and incubated for up to 9 days for TME studies. It was observed that NIH3T3 cells promoted the kinetics of tumor organoid formation. The two types of cells self-organized into core-shell chimeric tumor spheroids (CTSs) with fibroblasts confined to the shell and cancer cells localized to the core. Confocal microscopy analysis indicated that a type-I collagen network developed inside the CTS along with increased TGF-β1 and α-SMA proteins. The results were correlated with a significantly increased stiffness in 3D co-cultured CTS up to 52 kPa as compared to two-dimensional (2D) co-culture. CTS was more resistant to 5-FU (IC 50 = 14.0 ± 3.9 μM) and Regorafenib (IC 50 = 49.8 ± 9.9 μM) compared to cells grown under the 2D condition 5-FU (IC 50 = 12.2 ± 3.7 μM) and Regorafenib (IC 50 = 5.9 ± 1.9 μM). Targeted collagen homeostasis with Sclerotiorin led to damaged collagen structure and disrupted the type-I collagen network within CTS. Such a treatment significantly sensitized collagen-supported CTS to 5-FU (IC 50 = 4.4 ± 1.3 μM) and to Regorafenib (IC 50 = 0.5 ± 0.2 μM). In summary, the efficient formation of colon cancer CTSs in a μ-well 3D culture platform allows exploration of the desmoplastic TME. The novel role of intratumor collagen quality as a drug sensitization target warrants further investigation.

Targeted intracellular delivery of biomolecules and therapeutic cargo enables the controlled manipulation of cellular processes. Laser-based optoporation has emerged as a versatile, non-invasive technique that employs light-based transient physical disruption of the cell membrane and achieves high transfection efficiency with low cell damage. Testing of the delivery efficiency of optoporation-based techniques has been conducted on single cells in monolayers, but its applicability in three-dimensional (3D) cell clusters/spheroids has not been explored. Cancer cells grown as 3D tumor spheroids are widely used in anti-cancer drug screening and can be potentially employed for testing delivery efficiency. Towards this goal, we demonstrated the optoporation-based high-throughput intracellular delivery of a model fluorescent cargo (propidium iodide, PI) within 3D SiHa human cervical cancer spheroids. To enable this technique, nano-spiked core-shell gold-coated polystyrene nanoparticles (ns-AuNPs) with a high surface-to-volume ratio were fabricated. ns-AuNPs exhibited high electric field enhancement and highly localized heating at an excitation wavelength of 680 nm. ns-AuNPs were co-incubated with cancer cells within hanging droplets to enable the rapid aggregation and assembly of spheroids. Nanosecond pulsed-laser excitation at the optimized values of laser fluence (45 mJ cm -2 ), pulse frequency (10 Hz), laser exposure time (30 s), and ns-AuNP concentration (5 × 10 10 particles per ml) resulted in the successful delivery of PI dye into cancer cells. This technique ensured high delivery efficiency (89.6 ± 2.8%) while maintaining high cellular viability (97.4 ± 0.4%), thereby validating the applicability of this technique for intracellular delivery. The optoporation-based strategy can enable high-throughput single cell manipulation, is scalable towards larger 3D tissue constructs, and may provide translational benefits for the delivery of anti-cancer therapeutics to tumors.

The ability to deliver foreign molecules into a single living cell with high transfection efficiency and high cell viability is of great interest in cell biology for applications in therapeutic development, diagnostics, and drug delivery towards personalized medicine. Various physical delivery methods have long demonstrated the ability to deliver cargo molecules directly to the cytoplasm or nucleus and the mechanisms underlying most of the approaches have been extensively investigated. However, most of these techniques are bulk approaches that are cell-specific and have low throughput delivery. In comparison to bulk measurements, single-cell measurement technologies can provide a better understanding of the interactions among molecules, organelles, cells, and the microenvironment, which can aid in the development of therapeutics and diagnostic tools. To elucidate distinct responses during cell genetic modification, methods to achieve transfection at the single-cell level are of great interest. In recent years, single-cell technologies have become increasingly robust and accessible, although limitations exist. This review article aims to cover various microfluidic-based physical methods for single-cell intracellular delivery such as electroporation, mechanoporation, microinjection, sonoporation, optoporation, magnetoporation, and thermoporation and their analysis. The mechanisms of various physical methods, their applications, limitations, and prospects are also elaborated.

Circulation tumor cells (CTCs) play an important role in metastasis and highly correlate with cancer progression; thus, CTCs could be considered as a powerful diagnosis tool. Our previous studies showed that the number of CTCs could be utilized for recurrence prediction in colorectal cancer (CRC); however, the odds ratio was still lower than five. To improve prognosis in CRC patients, we analyzed CTC clusters/microemboli, CTC numbers, and carcinoembryonic antigen (CEA)/carbohydrate antigen 19-9 (CA19-9) levels using a self-assembled cell array (SACA) chip system for recurrence prediction. In CRC patients, the presence of CTC clusters/microemboli may have higher correlation in metastasis when compared to the high number of CTCs. Additionally, when both the number of CTCs and serum CEA levels are high, very high odds ratios of 24.4 and 17.1 are observed in patients at all stages and stage III of CRC, respectively. The high number of CTCs and CTC clusters/microemboli simultaneously suggests the high chance of relapse (odds ratio 8.4). Overall, the characteristic of CTC clusters/microemboli, CEA level, and CTC number have a clinical potential to enhance CRC prognosis.

Background: Due to the noninvasive nature of boron neutron capture therapy (BNCT), it is considered a promising cancer treatment method. Aim: To investigate whether polyvinyl alcohol/boric acid crosslinked nanoparticles (PVA/BA NPs) are an efficient delivery system for BNCT. Materials & methods: PVA/BA NPs were synthesized and cocultured with brain and oral cancers cells for BNCT. Results: PVA/BA NPs had a boron-loading capacity of 7.83 ± 1.75 w/w%. They accumulated in brain and oral cancers cells at least threefold more than in fibroblasts and macrophages. The IC 50  values of the brain and oral cancers cells were at least ninefold and sixfold lower than those of fibroblasts and macrophages, respectively. Conclusion: Theoretically, PVA/BA NPs target brain and oral cancers cells and could offer improved therapeutic outcomes of BNCT.

Bovine serum albumin (BSA)-encapsulated copper sulfide nanocrystals (CuS NCs) were prepared by heating an alkaline solution containing copper ions and BSA without an additional sulfur source. At a high BSA concentration (0.8 mM), nanoassembly of the as-formed CuS NCs occurs to form BSA-CuS NCs as a result of the formation of BSA gel-like structures. In addition to their intrinsic photothermal properties, the BSA-CuS NCs possess rich surface vacancies and thus exhibit enzyme-like and photodynamic activities. Spontaneous generation of hydrogen peroxide (H 2 O 2 ) led to the in situ formation of copper peroxide (CPO) nanodots on the BSA-CuS NCs to catalyze singlet oxygen radical generation. The antimicrobial response was enhanced by >60-fold upon NIR laser irradiation, which was ascribed to the combined effect of the photodynamic and photothermal inactivation of bacteria. Furthermore, BSA-CuS NCs were transdermally administered onto a methicillin-resistant Staphylococcus aureus -infected wound and eradicated >99% of bacteria in just 1 min under NIR illumination due to the additional peroxidase-like activity of BSA-CuS NCs, transforming H 2 O 2 at the infection site into hydroxyl radicals and thus increasing the synergistic effect from photodynamic and photothermal treatment. The BSA-CuS NCs exhibited insignificant in vitro cytotoxicity and hemolysis and thus can serve as highly biocompatible bactericides in preclinical applications to effectively eradicate bacteria.

The ability to deliver foreign cargos into single living cells is of great interest in cell biology and therapeutic research. Here, we have reported a single or multiple position based nano-localized single-cell nano-electroporation platform. The device consists of an array of triangular shape ITO nano-electrodes with a 70 nm gap between two nano-electrodes, each having a 40 nm tip diameter. The voltage is applied between nano-electrodes to generate an intense electric field, which electroporates multiple nano-localized regions of the targeted single-cell membrane, and biomolecules are gently delivered into cells by pressurizing pump flow, without affecting cell viability. The platform successfully delivers dyes, QDs, and plasmids into different cell types with the variation of field strength, pulse duration, and the number of pulses. This new approach allows us to analyze delivery of different biomolecules into single living cells with high transfection efficiency (>96%, for CL1-0 cells) and high cell viability (∼98%), which are potentially beneficial for cellular therapy and diagnostic purposes.

In this paper, a nanobiosensor with surface-enhanced Raman scattering (SERS) capability is introduced for highly sensitive miRNA detection in colorectal cancer. This sensor was designed and fabricated by employing a nanoshielding mechanism from nanopolystyrene beads to resist reactive ion etching and allow anisotropic electrochemical etching, producing high-aspect-ratio, surface-corrugated nanopillars (SiNPs) on a silicon wafer to create extensive hot spots along the nanopillars for improved SERS signals. SERS enhancements were correlated with nanorange roughness, indicating that hot spots along the pillars were the crucial factor to improve the SERS effect. We achieved the detection capability of a trace amount of R6G (10 -8 M), and the SERS signal enhancement factor (EF) was close to 1.0 × 10 7 on surface-corrugated gold SiNPs. miRNA samples were also demonstrated on this sensor with good sensitivity and specificity. The target molecule miR-21-Cy5 was easily monitored through Raman spectrum variation with a PCR-comparable concentration at around 100 pM with clear nucleotide-specific Raman signals, which is also suitable for biomolecule sensing.

We report infrared (IR) pulse laser-activated highly efficient parallel intracellular delivery by using an array of titanium microdish (TMD) device. Upon IR laser pulse irradiation, a two-dimensional array of TMD device generated photothermal cavitation bubbles to disrupt the cell membrane surface and create transient membrane pores to deliver biomolecules into cells by a simple diffusion process. We successfully delivered the dyes and different sizes of dextran in different cell types with variations of laser pulses. Our platform has the ability to transfect more than a million cells in a parallel fashion within a minute. The best results were achieved for SiHa cells with a delivery efficiency of 96% and a cell viability of around 98% for propidium iodide dye using 600 pulses, whereas a delivery efficiency of 98% and a cell viability of 100% were obtained for dextran 3000 MW delivery using 700 pulses. For dextran 10,000 MW, the delivery efficiency was 92% and the cell viability was 98%, respectively. The device is compact, easy-to-use, and potentially applicable for cellular therapy and diagnostic purposes.

Tissue-specific microenvironmental factors contribute to the targeting preferences of metastatic cancers. However, the physical attributes of the premetastatic microenvironment are not yet fully characterized. In this research, we develop a transwell-based alginate hydrogel (TAH) model to study how permeability, stiffness, and roughness of a hanging alginate hydrogel regulate breast cancer cell homing. In this model, a layer of physically characterized alginate hydrogel is formed at the bottom of a transwell insert, which is placed into a matching culture well with an adherent monolayer of breast cancer cells. We found that breast cancer cells dissociate from the monolayer and home to the TAH for continual growth. The process is facilitated by the presence of rich serum in the upper chamber, the increased stiffness of the gel, as well as its surface roughness. This model is able to support the homing ability of MCF-7 and MDA-MB-231 cells drifting across the vertical distance in the culture medium. Cells homing to the TAH display stemness phenotype morphologically and biochemically. Taken together, these findings suggest that permeability, stiffness, and roughness are important physical factors to regulate breast cancer homing to a premetastatic microenvironment.

The brain is an intricate network with complex organizational principles facilitating a concerted communication between single-neurons, distinct neuron populations, and remote brain areas. The communication, technically referred to as connectivity, between single-neurons, is the center of many investigations aimed at elucidating pathophysiology, anatomical differences, and structural and functional features. In comparison with bulk analysis, single-neuron analysis can provide precise information about neurons or even sub-neuron level electrophysiology, anatomical differences, pathophysiology, structural and functional features, in addition to their communications with other neurons, and can promote essential information to understand the brain and its activity. This review highlights various single-neuron models and their behaviors, followed by different analysis methods. Again, to elucidate cellular dynamics in terms of electrophysiology at the single-neuron level, we emphasize in detail the role of single-neuron mapping and electrophysiological recording. We also elaborate on the recent development of single-neuron isolation, manipulation, and therapeutic progress using advanced micro/nanofluidic devices, as well as microinjection, electroporation, microelectrode array, optical transfection, optogenetic techniques. Further, the development in the field of artificial intelligence in relation to single-neurons is highlighted. The review concludes with between limitations and future prospects of single-neuron analyses.

In this work, we aim to study zinc oxide (ZnO)-based functional materials over cotton fabrics and their effects after gamma ray exposure of 9 kGy. We found that the binding of the nanoparticles with cotton fabrics can be enhanced after irradiation. This could be due to the oxygen deficiency or defects created in the interface between ZnO and cotton fabrics after irradiation. Near-edge X-ray absorption fine structure and X-ray photoelectron spectroscopy (XPS) were used to detect the oxygen inadequacies generated in the interior and at the surface of the ZnO nanoparticles after gamma ray exposure. XPS results showed that the binding energy of Zn shifts by 2 eV at 1.5 kGy and by 4 eV at 9 kGy. This huge shift of about 4 eV is completely different from other works due to the reaction that takes place on the interface between ZnO nanostructures and cotton fabrics after gamma ray irradiation. Overall, this work suggests that after gamma ray irradiation, there is an enhanced level of binding between the coated functional nanoparticles and cotton fabrics, which can be advantageous for the textile industries., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 American Chemical Society.)

Here, an efficient intracellular delivery of molecules with high cell viability is reported using nanosecond-pulsed laser-activated plasmonic photoporation, mediated by high-aspect-ratio nano-corrugated mushroom-shaped gold-coated polystyrene nanoparticles (nm-AuPNPs) at near-infrared wavelength. Upon pulsed laser illumination, nm-AuPNPs exhibit greater plasmonic extinction than spherical AuPNPs, which increase their energy efficiency and reduce the necessary illumination of light, effectively controlling cell damage and improving the delivery efficiency. Nm-AuPNPs exhibit surface plasmon absorption at near infrared region with a peak at 945 nm. Pulsed laser illumination at this plasmon peak triggers explosive nanobubbles, which create transient membrane pores, allowing the delivery of dyes, quantum dots and plasmids into the different cell types. The results can be tuned by laser fluence, exposure time, molecular size and concentration of nm-AuPNPs. The best results are found for CL1-0 cells, which yielded a 94% intracellular PI dye uptake and ∼100% cell viability at 35 mJ cm-2 laser fluence for 945 nm wavelength. Thus, the presented approach has proven to have an inevitable potential for biological cell research and therapeutic applications.

We have prepared copper nanoclusters (Cu NCs) in the presence of bovine serum albumin (BSA) and 1,3-propanedithiol (PDT). The PDT/BSA-Cu NCs possess great activities against different types of bacteria, including non-multidrug-resistant bacteria (Escherichia coli, Salmonella Enteritidis, Pseudomonas aeruginosa, and Staphylococcus aureus) and multidrug-resistant bacteria (methicillin-resistant S. aureus). Their minimal inhibitory concentration (MIC) values are at least 242-fold and 10-fold lower than that of the free PDT and BSA-Cu NCs, respectively. The PDT/BSA-Cu NCs are strongly bound to the bacterial membrane, in which they induce the generation of ascorbyl (Asc) and perhydroxyl (HOO) radicals that result in disruption of their membrane integrity. At a concentration of 100-fold higher than their MIC for Escherichia coli, the PDT/BSA-Cu NCs exhibit negligible cytotoxicity towards the tested mammalian cells and show insignificant hemolysis. We have further demonstrated that low-cost PDT/BSA-Cu NCs-coated carbon fiber fabrics (CFFs) are effective against antibacterial growth, showing their great potential for antifouling applications., Competing Interests: Declaration of Competing Interest All authors have approved to the final version of the manuscript. The manuscript has not been previously published, in whole or in part, and is not under consideration by any other journal. We confirm that all authors are aware of and accept responsibility for the manuscript., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Aim: To improve the killing efficacy of head and neck squamous cells (SAS) by boric acid-mediated boron neutron capture therapy (BNCT).  Materials & methods:  Boric acid-containing chitosan/alginate/polyvinyl alcohol nanoparticles (B-capNPs) were manufactured using the nano-electrospray process.  Results: Less than 10% of the boric acid leaked from the B-capNPs over 2 days. The B-capNPs killed up to 2.8-fold more SAS cells and reduced cytotoxicity tenfold when compared with pure boric acid alone. B-capNPs show selective uptake in tumor cells with tumor/normal ratios of SAS to normal (NIH 3T3) and macrophage (RAW 264.7) cells of 4.0 and 3.5, respectively, which are greater than the minimum acceptable tumor/normal ratio for BNCT of 2.5. Conclusion: These findings illustrate that B-capNPs may be more superior as BNCT drugs than pure boric acid.

Multifunctional lanthanide-doped upconversion nanoparticles (UCNPs) have spread their wings in the fields of flexible optoelectronics and biomedical applications. One of the ongoing challenges lies in achieving UCNP-based nanocomposites, which enable a continuous-wave (CW) laser action at ultralow thresholds. Here, gold sandwich UCNP nanocomposites [gold (Au 1 )-UCNP-gold (Au 2 )] capable of exhibiting lasing at ultralow thresholds under CW excitation are demonstrated. The metastable energy-level characteristics of lanthanides are advantageous for creating population inversion. In particular, localized surface plasmon resonance-based electromagnetic hotspots in the nanocomposites and the huge enhancement of scattering coefficient for the formation of coherent closed loops due to multiple scattering facilitate the process of stimulated emissions as confirmed by theoretical simulations. The nanocomposites are subjected to stretchable systems for enhancing the lasing action (threshold ∼ 0.06 kW cm -2 ) via a light-trapping effect. The applications in bioimaging of HeLa cells and antibacterial activity (photothermal therapy) are demonstrated using the newly designed Au 1 -UCNP-Au 2 nanocomposites.

Electrodynamic systems for bioanalytical applications constantly suffer from biofouling due to electrical field-induced nonspecific bioadsorption on electrode surfaces. To minimize this issue, surface modification using anti-biofouling and conductive materials is necessary to not only protect the electrode surface from nonspecific bioadsorption but also maintain desired electrodynamic properties for electrode operation. In this study, we designed and prepared a conductive, zwitterionic, and self-doped sulfonated polyaniline (SPANI) coating on Au electrode surfaces for anti-biofouling applications. The zwitterionic coating was fabricated by electrochemical polymerization of aniline on the Au electrode surface functionalized with cysteamine (HS-CH 2 CH 2 -NH 2 ) and then a post-polymerization treatment with fuming sulfuric acid. We found that the SPANI-coated electrodes exhibited an excellent anti-biofouling ability in dielectrophoresis (DEP) capturing-and-releasing processes, with a very low average residual mass rate of 1.44% for the SPANI-5s electrode, whereas electrodes modified with poly(ethylene glycol) (PEG) gave an average residual mass rate of 14.30%. Even under continuous operation for more than 1 h, the SPANI-5s electrode still showed stable anti-biofouling ability for an 11-cycle E. coli capturing-and-releasing DEP process, with the residual mass rate for all 11 cycles being kept at or below 2.18% to give an average residual mass rate of 1.62% with a standard deviation of 0.40%. This study demonstrates that electrodynamic systems with zwitterionic SPANI coated on open electrode surfaces can excellently function with decent conductance and anti-biofouling performance.

A microfluidic chip, which can separate and enrich leukocytes from whole blood, is proposed. The chip has 10 switchback curve channels, which are connected by straight channels. The straight channels are designed to permit the inertial migration effect and to concentrate the blood cells, while the curve channels allow the Dean flow to further classify the blood cells based on the cell sizes. Hydrodynamic suction is also utilized to remove smaller blood cells (e.g., red blood cell (RBC)) in the curve channels for higher separation purity. By employing the inertial migration, Dean flow force, and hydrodynamic suction in a continuous flow system, our chip successfully separates large white blood cells (WBCs) from the whole blood with the processing rates as high as 1 × 10 8 cells/sec at a high recovery rate at 93.2% and very few RBCs (~0.1%)., Competing Interests: The authors declare no conflict of interest.

Early diagnosis of bacterial infections is crucial to improving survival rates by enabling treatment with appropriate antibiotics within the first few hours of infection. This paper presents a highly sensitive amperometric biosensor for the detection of several pathogenic bacterial cells in blood plasma around 30 min. The proposed device is based on an electropolymerized self-assembled layer on gold nanoparticles operated in a portable nano-sieving microfluidic system (NS-MFS). The redox-active gold nanoparticles (raGNPs) enhanced the electrical conductivity and provided a greater number of electrochemically active molecules for sensing, while improving resistance to the fouling of sensors by oxidation products in blood plasma. The detection limit of the device has been shown to reach 10 CFU/mL for Pseudomonas aeruginosa and Staphylococcus aureus spiked in plasma. The dynamic range of the sensing system falls between 10 and 10 5 CFU/mL in a buffer solution by cyclic voltammetry (CV) measurements. The results demonstrated that the raGNPs/NS-MFS can successful detect P. aeruginosa and S. aureus in human plasma, and is very useful for the diagnosis of bacteremia from clinical samples., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Stable and low-cost carbon dots (C-dots) were prepared from polyethylenimine (PEI) by a hydrothermal method. It is found that the fluorescence of the C-dots (best measured at excitation/emission wavelengths of 365/473 nm) is quenched by selective oxidation of surface PEI by periodate but recovers in the presence of uric acid (UA). It is assumed that this is due to the selective reduction of the nitrone groups to hydroxylamine groups by UA. The findings were used to design a fluorometric method for determination of UA that has a 2.3 nM detection limit. This is lower than that of reported fluorometric and enzymatic assays. The performance of the method has been validated by determination of UA in samples of human saliva. It is found that the results agree well with those obtained by a commercial UA assay. Graphical abstract Schematic presentation of the polyethylenimine (PEI) carbon nanodots (C-dots) as a fluorescent probe for uric acid. Their fluorescence is quenched by periodate (IO 4 - ) due to oxidative formation of nitrone groups, an subsequently restored due to reduction by uric acid (UA).

Colorectal cancer (CRC) is the second most common cause of cancer-related death worldwide. Detecting and enumerating circulating tumor cells (CTCs) in patients with colorectal cancer emerged as an important prognostic tool which provides a direct estimate of metastatic potential. Improving the turnaround time and decreasing sample volume is critical for incorporating this liquid biopsy tool into routine practice. The objective of the current study was to validate the clinical feasibility of a self-assembled cell array (SACA) chip, a CTC counting platform with less than 4 h turnaround time, in patients with newly diagnosed colorectal cancers. In total, 179 patients with newly diagnosed colorectal cancers from a single institute were enrolled. Epithelial cell adhesion molecule positive (EpCAM(+)), cluster of differentiation 45 negative (CD45(-)) cells were isolated and enumerated from 2 mL of peripheral vein blood (PB) and inferior mesenteric vein blood (IMV) samples obtained during surgery. We found that the CTC count in PB but not IMV correlates with disease stages. Neoadjuvant chemotherapy did not lead to decreased CTC count in both types of blood samples. With cutoffs of four CTCs per 2 mL of blood, and serum carcinoembryonic antigen (CEA) level of 5 ng/mL, patients with non-metastatic disease were more likely to experience recurrence if they had high PB CTC count and high serum CEA concentration (odds ratio, 8.9). Our study demonstrates the feasibility of enumerating CTCs with a SACA chip in patients with colorectal cancer., Competing Interests: The authors declare no conflicts of interest.

The last 20 years have seen the blooming of microfluidics technologies applied to biological sciences. Microfluidics provides effective tools for biological analysis, allowing the experimentalists to extend their playground to single cells and single molecules, with high throughput and resolution which were inconceivable few decades ago. In particular, microfluidic devices are profoundly changing the conventional way of studying the cell motility and cell migratory dynamics. In this chapter we will furnish a comprehensive view of the advancements made in the research domain of confinement-induced cell migration, thanks to the use of microfluidic devices. The chapter is subdivided in three parts. Each section will be addressing one of the fundamental questions that the microfluidic technology is contributing to unravel: (i) where cell migration takes place, (ii) why cells migrate and, (iii) how the cells migrate. The first introductory part is devoted to a thumbnail, and partially historical, description of microfluidics and its impact in biological sciences. Stress will be put on two aspects of the devices fabrication process, which are crucial for biological applications: materials used and coating methods. The second paragraph concerns the cell migration induced by environmental cues: chemical, leading to chemotaxis, mechanical, at the basis of mechanotaxis, and electrical, which induces electrotaxis. Each of them will be addressed separately, highlighting the fundamental role of microfluidics in providing the well-controlled experimental conditions where cell migration can be induced, investigated and ultimately understood. The third part of the chapter is entirely dedicated to how the cells move in confined environments. Invadosomes (the joint name for podosomes and invadopodia) are cell protrusion that contribute actively to cell migration or invasion. The formation of invadosomes under confinement is a research topic that only recently has caught the attention of the scientific community: microfluidic design is helping shaping the future direction of this emerging field of research.

The investigation of human disease mechanisms is difficult due to the heterogeneity in gene expression and the physiological state of cells in a given population. In comparison to bulk cell measurements, single-cell measurement technologies can provide a better understanding of the interactions among molecules, organelles, cells, and the microenvironment, which can aid in the development of therapeutics and diagnostic tools. In recent years, single-cell technologies have become increasingly robust and accessible, although limitations exist. In this review, we describe the recent advances in single-cell technologies and their applications in single-cell manipulation, diagnosis, and therapeutics development.