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1.
Article in English | MEDLINE | ID: mdl-38905518

ABSTRACT

Herein we report the assessment of the effects of shockwave (SW) impacts on adult rat hippocampal progenitor cell (AHPC) neurospheres (NSs), which are used as in vitro brain models, for enhancing our understanding of the mechanisms of traumatic brain injury (TBI). The assessment has been achieved by using culture dishes and a new microchip. The microchip allows the chemicals released from the brain models cultured inside the cell culture chamber under SW impacts to diffuse to the nanosensors in adjacent sensor chambers through built-in diffusion barriers, which are used to prevent the cells from entering the sensor chambers, thereby mitigating the biofouling issues of the sensor surface. Experiments showed the negative impact of the SW on the viability, proliferation, and differentiation of the cells within the NSs. A qPCR gene expression analysis was performed and appeared to confirm some of the immunocytochemistry (ICC) results. Finally, we demonstrated that the microchip can be used to monitor lactate dehydrogenase (LDH) released from the AHPC-NSs subjected to SW impacts. As expected, LDH levels changed when AHPC-NSs were injured by SW impacts, verifying this chip can be used for assessing the degrees of injuries to AHPC-NSs by monitoring LDH levels. Taken together, these results suggest the feasibility of using the chip to better understand the interactions between SW impacts and in vitro brain models, paving the way for potentially establishing in vitro TBI models on a chip.

2.
ACS Sens ; 8(10): 3701-3712, 2023 10 27.
Article in English | MEDLINE | ID: mdl-37738233

ABSTRACT

Integrin tensions are critical for cell mechanotransduction. By converting force to fluorescence, molecular tension sensors image integrin tensions in live cells with a high resolution. However, the fluorescence signal intensity results collectively from integrin tension magnitude, tension dwell time, integrin density, sensor accessibility, and so forth, making it highly challenging to specifically monitor the molecular force level of integrin tensions. Here, a ratiometric tension sensor (RTS) was developed to exclusively monitor the integrin tension magnitude. The RTS consists of two tension-sensing units that are coupled in series and always subject to the same integrin tension. These two units are activated by tension to fluoresce in separate spectra and with different activation rates. The ratio of their activation probabilities, reported by fluorescence ratiometric measurement, is solely determined by the local integrin tension magnitude. RTS responded sensitively to the variation of integrin tension magnitude in platelets and focal adhesions due to different cell plating times, actomyosin inhibition, or vinculin knockout. At last, RTS confirmed that integrin tension magnitude in platelets and focal adhesions decreases monotonically with the substrate rigidity, verifying the rigidity dependence of integrin tensions in live cells and suggesting that integrin tension magnitude could be a key biomechanical factor in cell rigidity sensing.


Subject(s)
Integrins , Mechanotransduction, Cellular , Integrins/analysis , Integrins/metabolism , Focal Adhesions/metabolism , Mechanical Phenomena , Actin Cytoskeleton/metabolism
3.
ACS Sens ; 8(6): 2255-2262, 2023 06 23.
Article in English | MEDLINE | ID: mdl-37276452

ABSTRACT

Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification technique that has been widely used for the detection of pathogens in many organisms. Current LAMP-based sensors usually require the LAMP products to be labeled in order for them to be detected. Here, we present a novel label-free LAMP chip, which consists of a nanopore thin-film sensor embedded inside a LAMP reaction chamber. A fraction of LAMP primers is immobilized on the sensor surface, allowing the LAMP products to be synthesized and bound to the sensor surface via immobilized primers. After the LAMP reaction components are removed from the reaction chamber, the amplified LAMP products bound to the sensor surface give rise to significantly increased transducing signals, which can be measured by a portable optical spectrometer through an optical fiber probe. As a demonstration, we used the LAMP chip to detect the causal agent of late blight, Phytophthora infestans, which is one of the most devastating plant pathogens and poses a major threat to sustainable crop production worldwide. We show that this chip can detect as low as 1 fg/µL of P. infestans DNA in 30 min, which corresponds to an attomolar level of 1.6 × 10-6 attomole/µL and is at least 10 times more sensitive than the currently available methods. This label-free sensing technology holds great promise to open up a new avenue for ultrasensitive, highly specific, rapid, and cost-effective point-of-care diagnostics of plant, animal, human, and foodborne pathogens.


Subject(s)
Nucleic Acid Amplification Techniques , Animals , Humans , Nucleic Acid Amplification Techniques/methods , DNA Primers/genetics
4.
ACS Appl Mater Interfaces ; 15(9): 12202-12208, 2023 Mar 08.
Article in English | MEDLINE | ID: mdl-36808523

ABSTRACT

We report a method to fabricate silicon micro-nanostructures of different shapes by tuning the number of layers and the sizes of self-assembled polystyrene beads, which serve as the mask, and by tuning the reactive ion etching (RIE) time. This process is simple, scalable, and inexpensive without using any sophisticated nanomanufacturing equipment. Specifically, in this work, we demonstrate the proposed process by fabricating silicon micro- or nanoflowers, micro- or nanobells, nanopyramids, and nanotriangles using a self-assembled monolayer or bilayer of polystyrene beads as the mask. We also fabricate flexible micro-nanostructures by using silicon molds with micro-nanostructures. Finally, we demonstrate the fabrication of bandage-type electrochemical sensors with micro-nanostructured working electrodes for detecting dopamine, a neurotransmitter related to stress and neurodegenerative diseases in artificial sweat. All these demonstrations indicate that the proposed process provides a low-cost, easy-to-use approach for fabricating silicon micro-nanostructures and flexible micro-nanostructures, thus paving a way for developing wearable micro-nanostructures enabled sensors for a variety of applications in an efficient manner.

5.
Lab Chip ; 23(6): 1649-1663, 2023 03 14.
Article in English | MEDLINE | ID: mdl-36751868

ABSTRACT

To improve our understanding of how the central nervous system functions in health and disease, we report the development of an integrated chip for studying the effects of the neurotransmitters dopamine and serotonin on adult rat hippocampal progenitor cell (AHPC) neurospheroids. This chip allows dopamine or serotonin located in one chamber to diffuse to AHPC neurospheroids cultured in an adjacent chamber through a built-in diffusion barrier created by an array of intentionally misaligned micropillars. The gaps among the micropillars are filled with porous poly(ethylene glycol) (PEG) gel to tune the permeability of the diffusion barrier. An electrochemical sensor is also integrated within the chamber where the neurospheroids can be cultured, thereby allowing monitoring of the concentrations of dopamine or serotonin. Experiments show that concentrations of the neurotransmitters inside the neurospheroid chamber can be increased over a period of several hours to over 10 days by controlling the compositions of the PEG gel inside the diffusion barrier. The AHPC neurospheroids cultured in the chip remain highly viable following dopamine or serotonin treatment. Cell proliferation and neuronal differentiation have also been observed following treatment, revealing that the AHPC neurospheroids are a valuable in vitro brain model for neurogenesis research. Finally, we show that by tuning the permeability of diffusion barrier, we can block transfer of Escherichia coli cells across the diffusion barrier, while allowing dopamine or serotonin to pass through. These results suggest the feasibility of using the chip to better understand the interactions between microbiota and brain via the gut-brain axis.


Subject(s)
Dopamine , Microfluidics , Rats , Animals , Serotonin , Brain , Neurotransmitter Agents
6.
Lab Chip ; 21(16): 3128-3136, 2021 08 21.
Article in English | MEDLINE | ID: mdl-34180491

ABSTRACT

Integrins are key players in platelet adhesion and aggregation. Integrin molecular tensions, the forces transmitted by integrin molecules, are regulated by both mechanical and biochemical cues, and the outside-in and inside-out signaling has been extensively studied. While the mechanical properties of platelets at static status have been studied by atomic force microscopy, traction force microscopy and tension sensors, the biomechanical properties of flowing platelets remain elusive. Herein, we report microfluidic chips grafted with integrin tension sensors for microfluidic-force mapping in platelets. Specifically, the process of integrin αIIbß3 mediating tension transmission and platelet adhesion under low flow rates has been obtained, and the process of platelet clustering at post-stenotic regions has been demonstrated. We found that flowing shear force can postpone the integrin-mediated tension transmission and platelet adhesion. We further evaluated the effect of Y-27632, a ROCK inhibitor that has been proven to reduce integrin-mediated platelet adhesion, at a series of concentrations and demonstrated that microfluidic chips with integrin tension sensors are sensitive to the concentration-dependent effects of Y-27632. Given their low cost and scalable throughput, these chips are ideal technical platforms for biological studies of platelets at flowing status and for platelet inhibitor or potential antiplatelet drug screening.


Subject(s)
Blood Platelets , Integrins , Microfluidics , rho-Associated Kinases/antagonists & inhibitors , Animals , Dogs , Platelet Adhesiveness , Stress, Mechanical
7.
Biosens Bioelectron ; 182: 113179, 2021 Jun 15.
Article in English | MEDLINE | ID: mdl-33774433

ABSTRACT

Glial cell-derived neurotrophic factor (GDNF) is a small protein potently promoting the survival of dopaminergic and motor neurons. GDNF can be secreted from different types of cells including the dopaminergic neural cell line, N27. N27 cells, a rat dopaminergic neural cell line, is regarded as a suitable in vitro model for Parkinson's disease (PD) research. For PD treatment, transcranial magnetic stimulation (TMS), a noninvasive therapeutic method, showed beneficial clinical effects, but the mechanism for its benefit is not understood. Because GDNF is a potent neurotrophic factor, it is of great value to evaluate if GDNF secretion from N27 cells can be affected by magnetic stimulation (MS). However, the current methods for detecting GDNF are time-consuming and expensive. In this paper we outline the detection of GDNF secretion from N27 cells by ultrasensitive nanopore thin film sensors (nanosensor) for the first time. As low as 2 pg/mL GDNF can be readily detected by the nanosensor. Furthermore, we show that MS can promote GDNF secretion from N27 cells. Specifically, the GDNF concentration in N27 cell-conditioned media under MS treatment shows statistically significant increase up to 2-fold after 5 days in vitro in comparison with the control. This nanosensor along with the in vitro PD model N27 cells provides a low-cost, easy-to-use, sensitive approach for studying potential cell biological mechanisms of the clinical benefits of MS on PD.


Subject(s)
Biosensing Techniques , Parkinson Disease , Animals , Dopamine , Glial Cell Line-Derived Neurotrophic Factor , Magnetic Phenomena , Parkinson Disease/therapy , Rats
8.
ACS Appl Mater Interfaces ; 12(41): 45789-45795, 2020 Oct 14.
Article in English | MEDLINE | ID: mdl-32960561

ABSTRACT

Because of the physiological and anatomical constraints of the eye, ophthalmic drug delivery is challenging. When applied topically, less than 1% of administered ophthalmic drugs reach the aqueous humor. The delivery of a drug within an efficient therapeutic concentration, to the required site of action, for an extended period of time, is complicated. Herein, a novel type of contact lens device, with embedded microtubes as drug containers, is reported. This device can provide a simple, noninvasive, extended drug release up to 45 days with higher bioavailability and lower risk for adverse effects. Another unique feature of the device is the release of drug triggered by stretching of the contact lens, indicating the possibility for achieving a self-adaptive drug release device for treating glaucoma patients.


Subject(s)
Contact Lenses, Hydrophilic , Drug Delivery Systems , Fluoresceins/therapeutic use , Glaucoma/drug therapy , Ophthalmic Solutions/therapeutic use , Timolol/therapeutic use , Diffusion , Drug Liberation , Fluoresceins/administration & dosage , Humans , Intracranial Hypertension/drug therapy , Microtubules/chemistry , Ophthalmic Solutions/administration & dosage , Optical Imaging , Particle Size , Surface Properties , Timolol/administration & dosage
9.
Micromachines (Basel) ; 11(7)2020 Jun 28.
Article in English | MEDLINE | ID: mdl-32605280

ABSTRACT

Alzheimer's disease (AD), Parkinson's disease (PD) and glaucoma are all regarded as neurodegenerative diseases (neuro-DDs) because these diseases are highly related to the degeneration loss of functions and death of neurons with aging. The conventional diagnostic methods such as neuroimaging for these diseases are not only expensive but also time-consuming, resulting in significant financial burdens for patients and public health challenge for nations around the world. Hence early detection of neuro-DDs in a cost-effective and rapid manner is critically needed. For the past decades, some chip-based detection technologies have been developed to address this challenge, showing great potential in achieving point-of-care (POC) diagnostics of neuro-DDs. In this review, chip-based detection of neuro-DDs' biomarkers enabled by different transducing mechanisms is evaluated.

10.
Biosens Bioelectron ; 140: 111342, 2019 Sep 01.
Article in English | MEDLINE | ID: mdl-31153018

ABSTRACT

Salicylic acid (SA) is a phytohormone regulating immune responses against pathogens. SA and its derivatives can be found in diverse food products, medicines, cosmetics and preservatives. While salicylates have potential disease-preventative activity, they can also cause health problems to people who are hypersensitive. The current SA detection methods are costly, labor-intensive and require bulky instruments. In this study, a structure-switching aptamer-based nanopore thin film sensor was developed for cost-effective, rapid, sensitive and simple detection of SA in both buffer and plant extracts. SA is a challenging target for aptamer selection using conventional systemic evolution of ligands by exponential enrichment (SELEX) due to its small size and scarcity of reactive groups for immobilization. By immobilizing the SELEX library instead of SA and screening the library using a structure-switching SELEX approach, a high affinity SA aptamer was identified. The nanopore thin film sensor platform can detect as low as 0.1 µM SA. This is much better than the sensitivity of antibody-based detection method. This nanosensor also exhibited good selectivity among SA and its common metabolites and can detect SA in Arabidopsis and rice using only about 1 µl plant extracts within less than 30 min. The integration of SA aptamer and nanopore thin film sensor provides a promising solution for low-cost, rapid, sensitive on-site detection of SA.


Subject(s)
Aptamers, Nucleotide/chemistry , Biosensing Techniques/methods , Salicylic Acid/analysis , Arabidopsis/chemistry , Biosensing Techniques/economics , Nanopores/ultrastructure , Oryza/chemistry , Plant Extracts/chemistry , SELEX Aptamer Technique , Time Factors
11.
J Nanosci Nanotechnol ; 19(12): 7591-7595, 2019 12 01.
Article in English | MEDLINE | ID: mdl-31196265

ABSTRACT

Cost-effective single-walled carbon nanotubes (SWCNTs) and poly(3,4-ethylenedioxythiophene) nanocomposite-microwire (N-MWs) electronic biosensor was fabricated by microlithography and layer-by-layer nanoassembly. Using N-MWs, we not only can achieve accurate large-scale registration and integration of nanoscale materials into a functional device, but also can enhance the sensing surface area significantly, which can open great opportunities for the cost-effectively fabrication of ultrasensitive biosensors. Using the binding assay between Protein A and Rabbit Immunoglobulin G (IgG) as a model, measurements find that the N-MWs electronic biosensor, containing carbon nanotubes (CNTs) and conductive polymers, exhibits field effect and the resistance changes upon its binding to biomolecules. The effect of the dimensions of the N-MWs on the sensitivity of the sensor has also been evaluated.


Subject(s)
Biosensing Techniques , Nanocomposites , Nanotubes, Carbon , Animals , Bridged Bicyclo Compounds, Heterocyclic , Electronics , Polymers , Rabbits
12.
Biomed Microdevices ; 20(4): 84, 2018 09 18.
Article in English | MEDLINE | ID: mdl-30229312

ABSTRACT

The original version of this article unfortunately contained a mistake. One line indicating statistical significance was improperly placed in Fig. 5.

13.
ACS Appl Mater Interfaces ; 10(34): 28269-28278, 2018 Aug 29.
Article in English | MEDLINE | ID: mdl-30080968

ABSTRACT

Transcranial magnetic stimulation (TMS) is a noninvasive neuromodulation technique, an FDA-approved treatment method for various neurological disorders such as depressive disorder, Parkinson's disease, post-traumatic stress disorder, and migraine. However, information concerning the molecular/cellular-level mechanisms of neurons under magnetic simulation (MS), particularly at the single neural cell level, is still lacking, resulting in very little knowledge of the effects of MS on neural cells. In this paper, the effects of MS on the behaviors of neural cell N27 at the single-cell level on coverslip glass substrate and anodic aluminum oxide (AAO) nanoporous substrate are reported for the first time. First, it has been found that the MS has a negligible cytotoxic effect on N27 cells. Second, MS decreases nuclear localization of paxillin, a focal adhesion protein that is known to enter the nucleus and modulate transcription. Third, the effect of MS on N27 cells can be clearly observed over 24 h, the duration of one cell cycle, after MS is applied to the cells. The size of cells under MS was found to be statistically smaller than that of cells without MS after one cell cycle. Furthermore, directly monitoring cell division process in the microholders on a chip revealed that the cells under MS generated statistically more daughter cells in one average cell cycle time than those without MS. All these results indicate that MS can affect the behavior of N27 cells, promoting their proliferation and regeneration.


Subject(s)
Neurons , Cell Proliferation , Cell Survival , Nanopores , Transcranial Magnetic Stimulation
14.
Biomed Microdevices ; 20(3): 62, 2018 07 30.
Article in English | MEDLINE | ID: mdl-30062494

ABSTRACT

Cells communicate through the extracellular matrix (ECM) in many physiological and pathological processes. This is particularly important during cell migration, where cell communication can alter both the speed and the direction of migration. However, most cell culture systems operate with large volumes relative to cell numbers, creating low cell densities and diluting factors that mediate cell communication. Furthermore, they lack the ability to isolate single cells or small groups of cells. Droplet forming devices allow for an ability to embed single or small groups of cells into small volume segregated 3D environments, increasing the cell density to physiological levels. In this paper we show a microfluidic droplet device for fabricating 3D collagen-based microtissues to study breast cancer cell motility. MDA-MB-231 cells fail to spread and divide in small, thin chambers. Cell migration is also stunted as compared to thick 3D gels. However, larger chambers formed by a thicker devices promote cell spreading, cell division and faster migration. In the large devices, both cell-ECM and cell-cell interactions affect cell motility. Increasing collagen density decreases cell migration and increasing the number of cells per chamber increases cell migration speed. Furthermore, cells appear to sense both the ECM-chamber wall interface as well as other cells. Cells migrate towards the ECM-chamber interface if within roughly 150 µm, whereas cells further than 150 µm tend to move towards the center of the chamber. Finally, while cells do not show enhanced movement towards the center of mass of a cell cluster, their migration speed is more variable when further away from the cell cluster center of mass. These results show that microfluidic droplet devices can array 3D collagen gels and promote cell spreading, division and migration similar to what is seen in thick 3D collagen gels. Furthermore, they can provide a new avenue to study cell migration and cell-cell communication at physiologically relevant cell densities.


Subject(s)
Cell Communication , Cell Movement , Collagen/chemistry , Extracellular Matrix/chemistry , Gels/chemistry , Cell Line, Tumor , Humans , Lab-On-A-Chip Devices
15.
Opt Lett ; 43(13): 3057-3060, 2018 Jul 01.
Article in English | MEDLINE | ID: mdl-29957780

ABSTRACT

This Letter reports a method to significantly improve the optical resolution of the anodic aluminum oxide (AAO) nanopore thin film sensor based on multi-cavity Fabry-Perot interference. The newly designed sensor is fabricated by bonding a layer of transparent polymer thin film (pTF), which is polydimethylsiloxane (PDMS), to a transparent AAO thin film to form a flexible pTF-nanopore sensor. In comparison with the AAO nanopore thin film sensor, the pTF-nanopore sensor shows a much-improved quality (Q) factor and optical resolution. Typical thicknesses of a PDMS layer and an AAO layer of the pTF-nanopore sensor are 80 µm and 2 µm, respectively. The pTF-nanopore sensor used for angle detection shows a sensitivity of 0.4 nm/deg with a resolution of 0.2 deg. The pTF-nanopore sensor can also be used for temperature monitoring with a sensitivity of 0.2 nm/°C and a resolution of 1°C.

16.
Nanomedicine ; 14(6): 1845-1852, 2018 08.
Article in English | MEDLINE | ID: mdl-29857195

ABSTRACT

This paper reports the multiplexed monitoring of two promising biomarkers, beta-amyloid (Aß42) and total tau (T-tau), in both buffer and cerebrospinal fluid (CSF) for Alzheimer's disease (AD) using label-free optical nanosensors. It has been found that 7.8 pg/ml of Aß42 in buffer and 15.6 pg/ml of T-au in buffer can be readily detected with very good specificity. Based on our measurements, the purchased CSF itself contains Aß42, whose concentration is estimated to be about 400 pg/ml. Aß42 and T-tau in the mixtures of Aß42 and T-tau spiked in CSF have been detected successfully, indicating the feasibility of the optical nanosensors to detect these biomarkers in clinical samples. For the measurements, only a small amount (~1 µl) of the samples is required. This type of sensor is suitable for point-of-care application to diagnose the AD due to its low cost and ease-of-operation.


Subject(s)
Aluminum Oxide/chemistry , Alzheimer Disease/cerebrospinal fluid , Alzheimer Disease/diagnosis , Amyloid beta-Peptides/cerebrospinal fluid , Biomarkers/cerebrospinal fluid , tau Proteins/cerebrospinal fluid , Adult , Aged , Aged, 80 and over , Biosensing Techniques , Electrodes , Female , Humans , Male , Middle Aged , Young Adult
17.
Biosens Bioelectron ; 105: 36-41, 2018 May 15.
Article in English | MEDLINE | ID: mdl-29351868

ABSTRACT

This paper reports an aptamer-based nanopore thin film sensor for detecting theophylline in the buffer solution and complex fluids including plant extracts and serum samples. Compared to antibody-based detection, aptamer-based detection offers many advantages such as low cost and high stability at elevated temperatures. Experiments found that this type of sensor can readily detect theophylline at a concentration as low as 0.05µM, which is much lower than the detection limit of current lab-based equipment such as liquid chromatography (LC). Experiments also found that the aptamer-based sensor has good specificity, selectivity, and reasonable reusability with a significantly improved dynamic detection range. By using the same nanopore thin film sensors as the reference sensors to further mitigate the non-specific binding effect, the theophylline in plant extracts and serum has been detected. Only a small amount (~1µL) of plant extracts or serum samples is required to measure theophylline. Its low cost and ease-of-operation make this type of sensor suitable for point-of-care application to monitor the theophylline level of patients in real time.


Subject(s)
Aptamers, Nucleotide/chemistry , Biosensing Techniques/instrumentation , Bronchodilator Agents/analysis , Nanopores/ultrastructure , Theophylline/analysis , Vasodilator Agents/analysis , Animals , Biosensing Techniques/methods , Bronchodilator Agents/blood , Caffeine/chemistry , Cattle , Equipment Design , Limit of Detection , Plant Extracts/chemistry , Theobromine/chemistry , Theophylline/blood , Vasodilator Agents/blood
18.
Opt Express ; 25(16): 19391-19397, 2017 Aug 07.
Article in English | MEDLINE | ID: mdl-29041133

ABSTRACT

In this paper, a drug loading and release device fabricated using nanopore thin film and layer-by-layer (LbL) nanoassembly is reported. The nanopore thin film is a layer of anodic aluminum oxide (AAO), consisting of honeycomb-shape nanopores. Using the LbL nanoassembly process, the drug, using gentamicin sulfate (GS) as the model, can be loaded into the nanopores and the stacked layers on the nanopore thin film surface. The drug release from the device is achieved by immersing it into flowing DI water. Both the loading and release processes can be monitored optically. The effect of the nanopore size/volume on drug loading and release has also been evaluated. Further, the neuron cells have been cultured and can grow normally on the nanopore thin film, verifying its bio-compatibility. The successful fabrication of nanopore thin film device on silicon membrane render it as a potential implantable controlled drug release device.

19.
Biosens Bioelectron ; 77: 491-8, 2016 Mar 15.
Article in English | MEDLINE | ID: mdl-26457734

ABSTRACT

Currently used cancer marker for prostate adenocarcinoma (PC), serum prostate-specific antigen (PSA), greatly overestimates PC population. Patients with high PSA levels have to undergo unnecessary but physically painful and expensive procedure such as prostate biopsies repeatedly. The reliability of PC test can be greatly increased by finding a protein that is secreted selectively by malignant, but not normal, prostate cells. A recently discovered novel protein, referred as neuroendocrine marker (NEM), is secreted only by malignant prostate cells and released in blood circulation. Although NEM seems to be significantly more reliable based on the data obtained from a limited cohort, currently available NEM ELISA is not suitable for undertaking a large study. Therefore, the goal of the present study was to develop an alternative, label-free assay system that can reliably measure NEM and PSA in patient samples. Herein an optofluidic chip that can reliably detect PSA as well as NEM in patient samples has been developed. The optofluidic chip, which consists of arrayed nanopore-based sensors fabricated from anodic aluminum oxide (AAO) thin film, offers improved sensitivity upon the optimization of the concentration of the detector antibodies immobilized on the sensor surface. The results demonstrate that the chip is reliable, extremely sensitive and requires just 1 µl of patient serum (or even less) to measure PSA and NEM even in a non-cancer individual. Compared with the traditional ELISA for PSA, the nanopore-based sensor assay is 50-100 fold more sensitive, and offers many advantages such as elimination of labeled antigen, need for sophisticated equipment and highly trained individuals. These advantages, along with the low cost, should make the technology suitable for point-of-care application to screen elderly male populations for PC and to monitor the progress of patients undergoing PC treatment.


Subject(s)
Biomarkers, Tumor/blood , Immunoassay/instrumentation , Lab-On-A-Chip Devices , Prostate-Specific Antigen/blood , Prostatic Neoplasms/blood , Surface Plasmon Resonance/instrumentation , Equipment Design , Equipment Failure Analysis , Humans , Male , Membranes, Artificial , Nanopores/ultrastructure , Optical Devices , Prostatic Neoplasms/diagnosis , Reproducibility of Results , Sensitivity and Specificity
20.
Micromachines (Basel) ; 7(5)2016 May 05.
Article in English | MEDLINE | ID: mdl-30404258

ABSTRACT

Arrayed three-dimensional (3D) micro-sized tissues with encapsulated cells (microtissues) have been fabricated by a droplet microfluidic chip. The extracellular matrix (ECM) is a polymerized collagen network. One or multiple breast cancer cells were embedded within the microtissues, which were stored in arrayed microchambers on the same chip without ECM droplet shrinkage over 48 h. The migration trajectory of the cells was recorded by optical microscopy. The migration speed was calculated in the range of 3⁻6 µm/h. Interestingly, cells in devices filled with a continuous collagen network migrated faster than those where only droplets were arrayed in the chambers. This is likely due to differences in the length scales of the ECM network, as cells embedded in thin collagen slabs also migrate slower than those in thick collagen slabs. In addition to migration, this technical platform can be potentially used to study cancer cell-stromal cell interactions and ECM remodeling in 3D tumor-mimicking environments.

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