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The integration of microfluidics with biomedical research is confronted with considerable limitations due to its body materials. With high content of water, hydrogels own superior biocompatibility and degradability. Can hydrogels become another material choice for the construction of microfluidic chips, particularly biofluidics? The present review aims to systematically establish the concept of hydrogel-based microfluidic chips (HMCs) and address three main concerns: i) why choosing hydrogels? ii) how to fabricate HMCs?, and iii) in which fields to apply HMCs? It is envisioned that hydrogels may be used increasingly as substitute for traditional materials and gradually act as the body material for microfluidic chips. The modifications of conventional process are highlighted to overcome issues arising from the incompatibility between the construction methods and hydrogel materials. Specifically targeting at the "soft and wet" hydrogels, an efficient flowchart of "i) high resolution template printing; ii) damage-free demolding; iii) twice-crosslinking bonding" is proposed. Accordingly, a broader microfluidic chip concept is proposed in terms of form and function. Potential biomedical applications of HMCs are discussed. This review also highlights the challenges arising from the material replacement, as well as the future directions of the proposed concept. Finally, the authors' viewpoints and perspectives for this emerging field are discussed., (© 2020 Wiley-VCH GmbH.)

Procalcitonin (PCT) is a known protein biomarker clinically used for the early stages of sepsis diagnosis and therapy guidance. For its reliable determination, sandwich format magnetic bead-based immunoassays with two different electrochemical detection approaches are described: (i) disposable screen-printed carbon electrodes (SPE-C, on-drop detection); (ii) electro-kinetically driven microfluidic chips with integrated Au electrodes (EMC-Au, on-chip detection). Both approaches exhibited enough sensitivity (limit of detection (LOD) of 0.1 and 0.04 ng mL -1 for SPE-C and EMC-Au, respectively; cutoff 0.5 ng mL -1 ), an adequate working range for the clinically relevant concentrations (0.5-1000 and 0.1-20 ng mL -1 for SPE-C and EMC-Au, respectively), and good precision (RSD < 9%), using low sample volumes (25 µL) with total assay times less than 20 min. The suitability of both approaches was successfully demonstrated by the analysis of human serum and plasma samples, for which good recoveries were obtained (89-120%). Furthermore, the EMC-Au approach enabled the easy automation of the process, constituting a reliable alternative diagnostic tool for on-site/bed-site clinical analysis.

Minimal residual disease (MRD) offers a highly independent prognostic factor for leukemia patients. However, challenges confronting conventional MRD assays are high invasiveness, as well as limited detection sensitivity and clinical applicability. Inspired by the self-adaptive skeleton and multiple suckers or tendrils of climbing plants, a biomimetic Multivalent Aptamer Nanoclimber (MANC)-functionalized microfluidic chip (MANC-Chip) is reported for minimally invasive, highly sensitive and clinically applicable MRD detection in the peripheral blood of T-cell acute lymphoblastic leukemia patients. The MANCs are synthesized by a simple co-polymerization reaction. Due to their flexible structure and cooperative multivalent effect, MANCs dramatically enhance the binding affinity of aptamers targeting leukemia cells. A deterministic lateral displacement-patterned microfluidic chip is designed to further increase the collision probability between MANCs and leukemia cells. Benefiting from the synergistic effect of multivalent binding and enhanced collision, a high capture efficiency of 92.2% for leukemia cells is achieved. Moreover, the captured leukemia cells can be released with high efficiency of 88.9% and high viability of 93.8% via nuclease treatment prior to downstream analysis. Overall, the excellent features of MANC-Chip make it very useful for precise detection of MRD and better understanding of leukemia., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Directional droplet transportation without extra energy input remains a challenge in microfluidic biochips for clinical detections. Herein, inspired by the water-collecting behaviors on the cactus spine, we fabricate nanomaterial-based superwettable microspine (SMS) chips. The bioinspired SMS chips are capable of spontaneous and directional droplet transportation by synergistically combining geometric asymmetry and surface superhydrophilicity. Based on theoretical models, the gradient of the Laplace pressure arising from the geometric asymmetry of the SMS chip can dominate the directional transportation of the droplet, and the superhydrophilicity of the nanomaterial-based microspine can also contribute to the droplet self-transportation. The multimicrochannel SMS chips provide a simple and energy efficient technology to realize accurate detection of serum prostate-specific antigen (PSA) from prostate cancer patients, showing great potential as a biosensing platform for clinical applications. We believe that our bioinspired superwettable two-dimensional conical surface will offer effective means for the design of smart microfluidic devices and have great potential applications in multicomponent biosensing and clinical detection.

Introduction : In recent years, extensive research has been conducted on using exosomes as biomarkers for cancer detection. Exosomes are 40-150 nm-sized extracellular vesicles released by all cell types, including tumor cells. Exosomes are stable in body fluids due to their lipid bilayer member and often contain DNA, RNA, and proteins. These exosomes can be harvested from blood, plasma, serum, urine, or saliva and analyzed for tumor-relevant mutations. Thus, exosomes provide an alternative to current methods of tumor detection. Areas covered : This review discusses the use of exosomal diagnostics in various tumor types as well as their examination in various clinical trials. The authors also discuss the limitations of exosome-based diagnostics in the clinical setting and provide examples of several studies in which the development and usage of microfluidic chips and nano-sensing devices have been utilized to address these obstacles. Expert commentary : In recent years, exosomes and their contents have exhibited potential as novel tumor detection markers despite the labor involved in their harvest and isolation. Despite this, much work is being done to optimize exosome capture and analysis. Thus, their roles as biomarkers in the clinical setting appear promising.

Screens of cancer stem cells (CSCs)-specific agents present significant challenges to conventional cell assays due to the difficulty in preparing CSCs ready for drug testing. To overcome this limitation, developed is a microfluidic single-cell assay for screening breast cancer stem cell-specific agents. This assay takes advantage of the single-cell clone-forming capability of CSCs, which can be specifically inhibited by CSC-targeting agents. The single-cell assay is performed on a microfluidic chip with an array of 3840 cell-capturing units; the single-cell arrays are easily formed by flowing a cell suspension into the microchip. Achieved is a single cell-capture rate of ≈60% thus allowing more than 2000 single cells to be analyzed in a single test. Over long-term suspension culture, only a minority of cells survive and form tumorspheres. The clone-formation rate of MCF-7, MDA-MB-231, and T47D cells is 1.67%, 5.78%, and 5.24%, respectively. The clone-forming inhibition assay is conducted by exposing the single-cell arrays to a set of anticancer agents. The CSC-targeting agents show complete inhibition of single-cell clone formation while the nontargeting ones show incomplete inhibition effects. The resulting microfluidic single-cell assay with the potential to screen CSC-specific agents with high efficiency provides new tools for individualized tumor therapy., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

As an interface between the blood flow and vessel wall, endothelial cells (ECs) are exposed to hemodynamic forces, and the biochemical molecules released from ECs-blood flow interaction are important determinants of vascular homeostasis. Versatile microfluidic chips have been designed to simulate the biological and physiological parameters of the human vascular system, but in situ and real-time monitoring of the mechanical force-triggered signals during vascular mechanotransduction still remains a significant challenge. Here, such challenge is fulfilled for the first time, by preparation of a flexible and stretchable electrochemical sensor and its incorporation into a microfluidic vascular chip. This allows simulating of in vivo physiological and biomechanical parameters of blood vessels, and simultaneously monitoring the mechanically induced biochemical signals in real time. Specifically, the cyclic circumferential stretch that is actually exerted on endothelium but is hard to reproduce in vitro is successfully recapitulated, and nitric oxide signals under normal blood pressure, as well as reactive oxygen species signals under hypertensive states, are well documented. Here, the first integration of a flexible electrochemical sensor into a microfluidic chip is reported, therefore paving a way to evaluate in vitro organs by built-in flexible sensors., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Exosomes are secreted by most cell types and circulate in body fluids. Recent studies have revealed that exosomes play a significant role in intercellular communication and are closely associated with the pathogenesis of disease. Therefore, exosomes are considered promising biomarkers for disease diagnosis. However, exosomes are always mixed with other components of body fluids. Consequently, separation methods for exosomes that allow high-purity and high-throughput separation with a high recovery rate and detection techniques for exosomes that are rapid, highly sensitive, highly specific, and have a low detection limit are indispensable for diagnostic applications. For decades, many exosome separation and detection techniques have been developed to achieve the aforementioned goals. However, in most cases, these two techniques are performed separately, which increases operation complexity, time consumption, and cost. The emergence of microfluidics offers a promising way to integrate exosome separation and detection functions into a single chip. Herein, an overview of conventional and microfluidics-based techniques for exosome separation and detection is presented. Moreover, the advantages and drawbacks of these techniques are compared., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Various driving methods are developed in microfluidic technology to manipulate small amounts of liquid. Among them, the photocontrol method is highlighted for its advantages in contactless and accuracy control. However, the illumination method may induce risks of radiation damage, thermal damage, or chemical environment changes. To avoid these risks, a photothermal pneumatic microfluidic device (PPMD) containing a photothermal composite, an air chamber, and a tube is designed, bioinspired by the feeding process of female mosquitos. One way to drive liquid columns at a constant velocity higher than 1.0 cm s−1 in microtubes without interface modification is successfully obtained, and a method to separate out water–oil two‐phase liquid mixtures is proposed. The integration of several single PPMDs enables complex logical operations (AND and OR) toward precise control of microreactions. This work is promising in fields of microfluidic chips, pneumatic devices, liquid transfer, microreactions, and water–oil phase separation. [ABSTRACT FROM AUTHOR]

Alginate as a good drug delivery vehicle has excellent biocompatibility and biodegradability. In the ionic gelation process between alginate and Ca2+, the violent reaction is the absence of a well-controlled strategy in the synthesizing calcium alginate (CaA) microgels. In this study, a concentration-controlled microfluidic chip with central buffer flow was designed and 3D-printed to well-control the synthesis process of CaA microgels by the diffusion mixing pattern. The diffusion mixing pattern in the microfluidic chip can slow down the ionic gelation process in the central stream. The particle size can be influenced by channel length and flow rate ratio, which can be regulated to 448 nm in length and 235 nm in diameter. The delivery ratio of Doxorubicin (Dox) in CaA microgels are up to 90% based on the central stream strategy. CaA@Dox microgels with pH-dependent release property significantly enhances the cell killing rate against human breast cancer cells (MCF-7). The diffusion mixing pattern gives rise to well-controlled synthesis of CaA microgels, serving as a continuous and controllable production process for advanced drug delivery systems. [ABSTRACT FROM AUTHOR]

Cell migration is an important process involved in wound healing, tissue development, and so on. Many studies have been conducted to explore how certain chemicals and electric fields induce cell movements in specific directions, which are phenomena termed chemotaxis and electrotaxis, respectively. However, phototaxis, the directional migration of cells or organisms toward or away from light, is rarely investigated due to the difficulty of generating a precise and controllable light gradient. In this study, we designed and fabricated a microfluidic chip for simultaneously culturing cells and generating a blue light gradient for guiding cell migration. A concentration gradient was first established inside this chip, and by illuminating it with a blue light-emitting diode (LED), a blue light gradient was generated underneath. Cell migration in response to this light stimulus was observed. It was found that lung cancer cells migrated to the dark side of the gradient, and the intracellular reactive oxygen species (ROS) was proportional to the intensity of the blue light. [ABSTRACT FROM AUTHOR]

Continuous and real‐time sensoring has received much attention in biomarker monitoring, toxicity assessment, and therapeutic agent tracking. However, its on‐site application is seriously limited by several stubborn defects including liability to fouling, signal drifting, short service life, poor repeatability, etc. Additionally, most current methods require extra sample pretreatment, delaying timely acquisition of testing results. To address these issues, MXene‐Ti3C2Tx based screen‐printed electrode incorporated with a dialysis microfluidic chip is constructed for a direct and continuous multicomponent analysis of whole blood. Dual‐function of MXene is developed and allows for simultaneous quantification of different target compounds through one device. Importantly, ratiometric sensing tactic is easily implemented in the system, which greatly alleviates signal drifting. As a proof of concept, this novel sensor is applied in hemodialysis, and continuous assay of urea, uric acid, and creatinine levels in human blood is realized. This work paves a new path for 2D MXene in biomedical and sensing applications. Introduction of MXene nanosheets into microfluidic chips contributes significantly to stable and sensitive signal outputs. Particularly, MXene acts as both a carrier and electrocatalyst, providing sufficient sites for locating other agents, and rendering occurrence of electrochemical reactions. Such dual‐function, together with the elaborately designed testing chip platform and strategies, realizes simultaneous quantification of different target compounds through one device. [ABSTRACT FROM AUTHOR]

A multifunctional microfluidic platform was demonstrated to monitor the interaction between tumor cells and endothelial cells by integrating a three-dimensional (3D) cell culture unit with a protein detection unit. In such a chip, breast cancer cells MCF7 were seeded into the collagen to form a 3D tumor environment while human umbilical vein endothelial cells (HUVECs) are seeded in the channel next to the collagen matrix. Thus, an in situ growth of angiogenic sprouting can be visualized through fluorescence in the 3D collagen matrix after a coculture of MCF7 and HUVEC after 4 days, which cannot be observed in the 2D culture environment. On the other hand, gold@silver core-shell nanorods were used as surface-enhanced Raman scattering (SERS) immunoprobes for the detection of the secretion of cytokine (vascular endothelial growth factor, VEGF). The limit of detection of the VEGF is 100 pg/mL. Further, as LiCl and bevacizumab can act as a promoter and an inhibitor of VEGF, the dynamic change of the concentration of VEGF under the stimulation of them was monitored by SERS signals. Thus, this integrated SERS microfluidic platform creates opportunity for the fundamental research of interaction between tumors and endothelial cells.

Circulating tumor cells (CTCs) represent invasive tumor cell populations and provide a noninvasive solution to the clinical management and research of tumors. Characterization of CTCs at single-cell resolution enables the comprehensive understanding of tumor heterogeneity and may benefit the diagnosis and treatment of cancer patients. However, most efforts have been made on enumeration and detection of CTCs, while little focus has been directed to single-cell study. Herein, an integrated microfluidic platform for single-cell isolation and analysis was established. After validating this platform on lung cancer cell lines, we detected and isolated single CTCs from the peripheral blood samples of 20 cancer patients before and after one treatment cycle. Furthermore, we performed single-cell whole-exome DNA sequencing on a single CTC from the peripheral blood sample of a representative early stage lung cancer patient. Among the blood samples of 20 patients, 15 of them were positive for CTC detection (75.0% detectable rate). Single-cell analysis revealed detailed genetic variations of the CTC, while six new gene mutations were detected in both single CTC and surgical specimen. This study provides a useful tool for the isolation and analysis of single CTCs from peripheral blood samples, which not only facilitates the early diagnosis of cancers but also helps to unravel the genetic information of tumor at a single-cell level. © 2019 International Society for Advancement of Cytometry., (© 2019 International Society for Advancement of Cytometry.)

Microfluidic chip electrophoresis has been widely employed for separation of various biochemical species owing to its advantages of low sample consumption, low cost, fast analysis, high throughput, and integration capability. In this article, we reviewed the development of four different modes of microfluidics-based electrophoresis technologies including capillary electrophoresis, gel electrophoresis, dielectrophoresis, and field (electric) flow fractionation. Coupling detection schemes on microfluidic electrophoresis platform were also reviewed such as optical, electrochemical, and mass spectrometry method. We further discussed the innovative applications of microfluidic electrophoresis for biomacromolecules (nucleic acids and proteins), biochemical small molecules (amino acids, metabolites, ions, etc.), and bioparticles (cells and pathogens) analysis. The future direction of microfluidic chip electrophoresis was predicted., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

The fate of stem cells at the single cell level with limited communication with other cells is still unknown due to the lack of an efficient tool for highly accurate molecular detection. Moreover, the conditional sensitivity of biological experiments requires a sufficient number of parallel experiments to support a conclusion. In this work, a microfluidic single cell chip is designed for use with a protein chip to investigate the effect of hydroxyapatite (HAp) on the osteogenic differentiation of human adipose-derived stem cells (hADSCs) in situ at the single cell level. By successfully detecting secretory proteins in situ, it is found that the HAp nanorods enhance osteogenic differentiation at the single cell level. In the chip, the single cell seeding approach confirms the osteogenic differentiation of the hADSCs, which endocytoses HAp, by reducing the influence of the factors secreted by neighboring differentiating cells. Most importantly, more than 7000 microchambers provide a sufficient number of parallel experiments for statistical analysis, which ensure a high level of repeatability of the HAp nanorod-induced osteogenic differentiation. The microfluidic chip comprising single cell culture microchambers with in situ detection capability is a promising tool for research on cell behavior or cell fate at the single cell level., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Monodisperse polyvinyl alcohol (PVA) microspheres have been widely used for targeted drug delivery, embolization, and templates. However, the fast and facile fabrication of PVA microspheres with uniform size and internal structure and good sphericity remains a challenge. In this study, the PVA microspheres with uniformity in the size, shape, and internal structure were rapidly fabricated, using single-emulsion droplet templates by an on-chip approach. First, we designed a polydimethylsiloxane (PDMS) microfluidic chip integrated with three functional units, used for the droplet generation, mixing of reagents, and pre-curing of PVA microspheres, respectively. Then, we precisely controlled the generation of PVA aqueous droplets, mixing of reagents, and the gelation rate for the production of high-quality microspheres by adjusting the pH value, flow rate, and the channel structure. The prepared PVA microspheres are characterized with good sphericity, uniform internal structure, and narrow size distribution. The microspheres have good adsorption capacity and recyclability for small-molecule drugs, as demonstrated by the adsorption and desorption of methylene blue (MB). The adsorption behavior is well described by the pseudo-second-order model, and intraparticle diffusion is as fast as the external film diffusion.

Microfluidic devices are used in a myriad of biomedical applications such as cancer screening, drug testing, and point-of-care diagnostics. Three-dimensional (3D) printing offers a low-cost, rapid prototyping, efficient fabrication method, as compared to the costly—in terms of time, labor, and resources—traditional fabrication method of soft lithography of poly(dimethylsiloxane) (PDMS). Various 3D printing methods are applicable, including fused deposition modeling, stereolithography, and photopolymer inkjet printing. Additionally, several materials are available that have low-viscosity in their raw form and, after printing and curing, exhibit high material strength, optical transparency, and biocompatibility. These features make 3D-printed microfluidic chips ideal for biomedical applications. However, for developing devices capable of long-term use, fouling—by nonspecific protein absorption and bacterial adhesion due to the intrinsic hydrophobicity of most 3D-printed materials—presents a barrier to reusability. For this reason, there is a growing interest in anti-fouling methods and materials. Traditional and emerging approaches to anti-fouling are presented in regard to their applicability to microfluidic chips, with a particular interest in approaches compatible with 3D-printed chips. [ABSTRACT FROM AUTHOR]

Hydrogel structures equipped with internal microchannels offer more in vivo-relevant models for construction of tissues and organs in vitro. However, currently used microfabrication methods of constructing microfluidic devices are not suitable for the handling of hydrogel. This study presents a novel method of fabricating hydrogel-based microfluidic chips by combining the casting and bonding processes. A twice cross-linking strategy is designed to obtain a bonding interface that has the same strength with the hydrogel bulk, which can be applied to arbitrary combinations of hydrogels. It is convenient to achieve the construction of hydrogel structures with channels in branched, spiral, serpentine, and multilayer forms. The experimental results show that the combination of gelatin and gelatin methacrylate (GelMA) owns the best biocompatibility and can promote cell functionalization. Based on these, a vessel-on-a-chip system with vascular function in both physiological and pathological situations is established, providing a promising model for further investigations such as vascularization, vascular inflammation, tissue engineering, and drug development. Taken together, a facile and cytocompatible approach is developed for engineering a user-defined hydrogel-based chip that can be potentially useful in developing vascularized tissue or organ models., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Microfluidic chips allow the rapid production of a library of nanoparticles (NPs) with distinct properties by changing the precursors and the flow rates, significantly decreasing the time for screening optimal formulation as carriers for drug delivery compared to conventional methods. The batch-to-batch reproducibility which is essential for clinical translation is achieved by precisely controlling the precursors and the flow rate, regardless of operators. Poly (lactic-co-glycolic acid) (PLGA) is the most widely used Food and Drug Administration (FDA)-approved biodegradable polymers. Researchers often combine PLGA with lipids or amphiphilic molecules to assemble into a core/shell structure to exploit the potential of PLGA-based NPs as powerful carriers for cancer-related drug delivery. In this review, we discuss the advantages associated with microfluidic chips for producing PLGA-based functional nanocomplexes for drug delivery. These laboratory-based methods can readily scale up to provide sufficient amount of PLGA-based NPs in microfluidic chips for clinical studies and industrial-scale production., (Copyright © 2017. Published by Elsevier B.V.)

Circulating tumor cells (CTCs), a type of cancer cells that spreads from primary or metastatic tumors into the bloodstream, can lead to a new fatal metastasis. As a new type of liquid biopsy, CTCs have become a hot pursuit and detection of CTCs offers the possibility for early diagnosis of cancers, earlier evaluation of chemotherapeutic efficacy and cancer recurrence, and choice of individual sensitive anti-cancer drugs. The fundamental challenges of capturing and characterizing CTCs are the extremely low number of CTCs in the blood and the intrinsic heterogeneity of CTCs. A series of microfluidic devices have been proposed for the analysis of CTCs with automation capability, precise flow behaviors, and significant advantages over the conventional larger scale systems. This review aims to provide in-depth insights into CTCs analysis, including various nanomaterial-based microfluidic chips for the capture and detection of CTCs based on the specific biochemical and physical properties of CTCs. The current developmental trends and promising research directions in the establishment of microfluidic chips for the capture and detection of CTCs are also discussed., Competing Interests: Competing Interests: The authors have declared that no competing interest exists.

Recently, a new observation method for monitoring of pH gradients in microfluidic free-flow electrophoresis has emerged. It is based on the use of chip-integrated fluorescent or luminescent micro sensor layers. These are able to monitor pH gradients in miniaturized separations in real time and spatially resolved; this is particularly useful in isoelectric focusing. Here these multifunctional microdevices that feature continuous separation, monitoring, and in some instances other functionalities, are reviewed. The employed microfabrication procedures to produce these devices are discussed and the different pH sensor matrices that were integrated and their applications in the separation of different types of biomolecules. The procedures for obtaining spatially resolved information about the separated molecules and the pH at the same time and different detection modalities to achieve this such as deep UV fluorescence as well as time-resolved referenced pH sensing and the integration of a precolumn labeling step into these platforms are also highlighted., (© 2017 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.)

The arterial microenvironment plays a vital role in the pathology of atherosclerosis (AS). However, the interplay between the arterial microenvironment and atherogenesis remains unclear, partially due to the gap between cell culture and animal experiments. Addressing this problem, the present study reports a microfluidic AS model reconstituting early-stage AS. Physiological or AS-prone hemodynamic conditions are recapitulated on the model. The on-chip model recaptures the atherogenic responses of endothelial cells (ECs) in ways that the Petri dish could not. Significant cytotoxicity of a clinical anti-atherosclerotic drug probucol is discovered on the model, which does not appear on Petri dish but is supported by previous clinical evidence. Moreover, the anti-AS efficiency of platinum-nanoparticles (Pt-NPs) on the model shows excellent consistency with animal experiments. The early-stage AS model shows an excellent connection between Petri dish and animal experiments and highlights its promising role in bridging fundamental AS research, drug screening, and clinical trials., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Background: In the present study, we aimed to investigate the influence of lactate shuttling on the functional polarization and spatial distribution of transitional cell carcinoma of the bladder (TCCB) cells and macrophages., Methods: We designed a microfluidic coculture chip for real-time integrative assays. The effect of lactate shuttling on the re-education of macrophages by TCCB cells was explored by measuring the levels of NO using a total NO assay kit and by evaluating the protein expression of iNOS, p-NFkB-p65, Arg-1 and HIF-1α via cell immunofluorescence and western blotting. Additionally, we examined TCCB cell viability using acridine orange/ethidium bromide (AO/EB) and MitoTracker staining. Moreover, the concentration distributions of lactate and large signaling proteins in the culture chambers were measured using 4',6-diamidino-2-phenylindole (DAPI) and fluorescein isothiocyanate-dextran (FITC-dextran). Furthermore, the recruitment of macrophages and the influence of macrophages on BC metastasis were observed via light microscopy., Results: We confirmed that TCCB cells reprogrammed macrophages into an M2 phenotype. Moreover, lactate inhibited M1 polarization and induced M2 polarization of macrophages, but blockade of cancer cell-macrophage lactate flux significantly inhibited the re-education of macrophages by TCCB cells. In addition, lactate diffused faster and deeper than large signaling proteins in the microfluidic tumor microenvironment. Furthermore, lactate alone induced the migration of macrophages, and M1, but not M2, macrophages reduced the motility of TCCB cells., Conclusions: TCCB cells reprogrammed macrophages into an M2 phenotype in a manner that depended on cancer cell-TAM lactate flux. Furthermore, the lactate shuttle may be a determinant of the density of TAMs in tumor tissue.

A surface enhanced Raman scattering (SERS)-assisted 3D barcode chip has been developed for high-throughput biosensing. The 3D barcode is realized through joint 2D spatial encoding with the Raman spectroscopic encoding, which stores the SERS fingerprint information in the format of a 2D array. Here, the concept of SERS-assisted 3D barcode is demonstrated through multiplex immunoassay, where simultaneous detection of multiple targets in different samples has been achieved using a microfluidic platform. First, multiple proteins in different samples are spatially separated using a microfluidic patterned antibody barcode substrate, forming a 2D hybridization array. Then the SERS probes are used to identify and quantify the proteins. As different SERS probes are labeled with different Raman reporters, they could be employed as "SERS tags" to incorporate spectroscopic information into the 3D barcode. In this 3D barcode, the 2D spatial information helps to differentiate the samples and targets while the SERS information allows quantitative multiplex detection. It is found that the SERS-assisted 3D barcode chip can not only accomplish one-step multiplex detection within 30 min but also achieve an ultrasensitivity down to 10 fg mL(-1) (≈70 aM), which is expected to provide a promising tool for high-throughput biomedical applications., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Active recombinant proteins are used for studying the biological functions of genes and for the development of therapeutic drugs. Overexpression of recombinant proteins in bacteria often results in the formation of inclusion bodies, which are protein aggregates with non-native conformations. Protein refolding is an important process for obtaining active recombinant proteins from inclusion bodies. However, the conventional refolding method of dialysis or dilution is time-consuming and recovered active protein yields are often low, and a cumbersome trial-and-error process is required to achieve success. To circumvent these difficulties, we used controllable diffusion through laminar flow in microchannels to regulate the denaturant concentration. This method largely aims at reducing protein aggregation during the refolding procedure. This Commentary introduces the principles of the protein refolding method using microfluidic chips and the advantage of our results as a tool for rapid and efficient recovery of active recombinant proteins from inclusion bodies.