Optimizing Microfluidic Impedance Cytometry by Bypass Electrode Layout Design.

Microfluidic impedance cytometry (MIC) has emerged as a popular technique for single-cell analysis. Traditional MIC electrode designs consist of a pair of (or three) working electrodes, and their detection performance needs further improvements for microorganisms. In this study, we designed an 8-electrode MIC device in which the center pair was defined as the working electrode, and the connection status of bypass electrodes could be changed. This allowed us to compare the performance of layouts with no bypasses and those with floating or grounding electrodes by simulation and experiment. The results of detecting Φ 5 µm beads revealed that both the grounding and the floating electrode outperformed the no bypass electrode, and the grounding electrode demonstrated the best signal-to-noise ratio (SNR), coefficient of variation (CV), and detection sensitivity. Furthermore, the effects of different bypass grounding areas (numbers of grounding electrodes) were investigated. Finally, particles passing at high horizontal positions can be detected, and Φ 1 µm beads can be measured in a wide channel (150 µm) using a fully grounding electrode, with the sensitivity of bead volume detection reaching 0.00097%. This provides a general MIC electrode optimization technology for detecting smaller particles, even macromolecular proteins, viruses, and exosomes in the future.

A disposable paper-based electrochemical biosensor decorated by electrospun cellulose acetate nanofibers for highly sensitive bio-detection.

Paper-based electrochemical sensors have the characteristics of flexibility, biocompatibility, environmental protection, low cost, wide availability, and hydropathy, which make them very suitable for the development and application of biological detection. This work proposes electrospun cellulose acetate nanofiber (CA NF)-decorated paper-based screen-printed (PBSP) electrode electrochemical sensors. The CA NFs were directly collected on the PBSP electrode through an electrospinning technique at an optimized voltage of 16 kV for 10 min. The sensor was functionalized with different bio-sensitive materials for detecting different targets, and its sensing capability was evaluated by CV, DPV, and chronoamperometry methods. The test results demonstrated that the CA NFs enhanced the detection sensitivity of the PBSP electrode, and the sensor showed good stability, repeatability, and specificity (p < 0.01, N = 3). The electrochemical sensing of the CA NF-decorated PBSP electrode exhibited a short detection duration of ∼5-7 min and detection ranges of 1 nmol mL-1-100 µmol mL-1, 100 fg mL-1-10 µg mL-1, and 1.5 × 102-106 CFU mL-1 and limits of detection of 0.71 nmol mL-1, 89.1 fg mL-1, and 30 CFU mL-1 for glucose, Ag85B protein, and E. coli O157:H7, respectively. These CA NF-decorated PBSP sensors can be used as a general electrochemical tool to detect, for example, organic substances, proteins, and bacteria, which are expected to achieve point-of-care testing of pathogenic microorganisms and have wide application prospects in biomedicine, clinical diagnosis, environmental monitoring, and food safety.

Direct, ultrafast, and sensitive detection of environmental pathogenic microorganisms based on a graphene biosensor.

Pathogenic microorganisms in the environment pose a serious threat to global human health. This study developed a reduced graphene oxide (rGO)-field effect transistor (FET) biosensor to realize the rapid and sensitive detection of pathogenic microorganisms. The rGO-FET sensors were prepared by in-situ thermal reduction method, and biorecognition elements were immobilized using a crosslinking agent to realize the surface functionalization of rGO. The rGO-FET biosensors can detect Escherichia coli O157:H7 as low as 1.4 CFU mL-1 within 46 s. The normalized current response was linearly correlated with E. coli concentration in the range of 1.4-1.4 × 107 CFU mL-1. The normalized current response of E. coli O157:H7 was about an order of magnitude higher than those of other microorganisms, indicating that the biosensor has good specificity. The current loss rates of the unmodified rGO-FET sensors and the biosensors modified with anti-E. coli O157:H7 after 30 days of storage at 4 °C were approximately 8% and 15%, respectively. Most importantly, the rGO-FET biosensors can directly detect real samples without pretreatment. Compared with other technologies, the rGO-FET biosensors can detect pathogenic microorganisms with a wider linear range in a shorter time, which is of great importance for the rapid warning and control of pathogenic microorganisms in the environment.

A self-contained and integrated microfluidic nano-detection system for the biosensing and analysis of molecular interactions.

Traditional detection methods have shortcomings such as time-consumption and requirement of large instruments, which cannot meet the demands for on-site detection or analysis. Silicon nanowire-field-effect transistor (SiNW-FET) biosensors have the advantages of high speed, high sensitivity, strong specificity, and ease of integration. However, SiNW-FET biosensors also have some demerits: they are too sensitive, environmental factors such as light, temperature, and pH easily cause interference, and their performance uniformity needs to be calibrated in advance. In this work, we constructed a self-contained and integrated microfluidic nano-detection system containing a SiNW-FET biosensor for bio-detection and analysis. All analysis processes including liquid sample delivery, optical modulation, constant temperature control, signal amplification and data acquisition, and result display were automatically performed. In series tests including light-guided ones by analyzing various types of samples with an automatic sample injection mode, the system shows good stability and robustness. Its signal accuracy was verified using a commercial high-precision ammeter (R2 = 0.9988), too. The feasibility of the system for bio-detection was verified using simulant samples of the typical microorganism Mycobacterium tuberculosis with a limit of detection of 1.0 fg mL-1. Furthermore, the process of the binding-dissociation of antibody-protein pairs was analyzed using the system, demonstrating the potential for molecular interaction analysis. This system is highly integrated, small in size, and easy to carry, which will be developed into a portable device for on-site bio-detection and analysis of molecular interactions to enable environmental testing, medical research, food and agricultural safety, military medicine, etc.

Rapid detection of airborne protein from Mycobacterium tuberculosis using a biosensor detection system.

Tuberculosis (TB), caused by infection with airborne Mycobacterium tuberculosis (MTB), seriously threatens human health and has become a public health problem of worldwide concern. To achieve effective control of TB, rapid and sensitive detection of MTB is particularly important. At present, the common detection methods for MTB cannot meet the requirements of speed, flexibility and portability simultaneously. In this work, a multichannel microfluidic chip was developed and packaged with an ultra-sensitive silicon nanowire field-effect-transistor biosensor. The fluid system was tested and optimized through simulation, and the best conditions were determined: the flow rate was 0.3 mL min-1 and the flow direction was perpendicular to a silicon nanowire. A one-way valve, a switching valve and a peristaltic pump were combined to establish a biosensor detection system to realize the automatic detection of TB samples. Then we systematically explained the factors affecting simulated exhaled breath condensate (SEBC) collection, and established and optimized the method for collection of SEBC from the perspective of collection volume and biological activity. The best collection conditions were determined for a 5 mm pipe diameter at 0 °C, and a sufficient sample volume was obtained in only 2 minutes for microfluidic detection. Then, the actual application value of the established collection method was further evaluated. Volunteers were recruited and this method was used to collect their exhaled breath condensate to analyze the collection effect. The system detected MTB in SEBC with good sensitivity (∼4 × 104 particles per mL). It is expected to be further integrated and miniaturized in the future to realize point-of-care testing.

Portable immunosensor directly and rapidly detects Mycobacterium tuberculosis in sputum.

Tuberculosis (TB) remains a public health problem that cannot be ignored. The portable and efficient detection of Mycobacterium tuberculosis (MTB) is important for the effective control of this disease. However, current detection techniques do not meet the requirements for MTB detection in the actual environment and often require cumbersome detection steps that are time consuming and inflexible. In this study, a portable immunosensor to detect MTB in sputum was prepared and then subjected to interface characterizations, such as scanning electron microscopy, hydrophilic angle test, and fluorescence characterization. The source and gate voltage of the device were optimized and tested using a non-contact photoresponse. The results showed that the sensitivity of the sensor to luminance increases with the decrease in source voltage. The gate voltage can substantially improve the response of the immunosensor to the normalized current of protein and amplify the signal at least 1.6 times. The optimal voltage detection conditions of source voltage (0.3 V) and gate voltage (0.1 V) were also determined. Several common proteins present in simulated saliva were used for anti-interference tests, and the sensor exhibited good specificity. Finally, the dilution gradient of an actual TB sputum sample was optimized. In the absence of preconditioning, a double-blind experiment was used to distinguish between the sputum from patients with TB and healthy individuals to shorten the TB detection time to a few minutes. Compared with the hospital's conventional detection method using cultures, the proposed method can complete the detection in a shorter time. This study provides a new strategy for the portable diagnosis of TB.

Rapid and Sensitive Detection of Mycobacterium tuberculosis by an Enhanced Nanobiosensor.

Tuberculosis (TB) mostly spreads from person to person through Mycobacterium tuberculosis (MTB). However, the majority of conventional detection methods for MTB cannot satisfy the requirements for actual TB detection. As one of the most promising powerful platforms, a silicon nanowire field-effect transistor (SiNW-FET) biosensor shows good prospect in TB detection. In this study, an enhanced SiNW-FET biosensor was developed for the rapid and sensitive detection of MTB. The surface functional parameters of the biosensor were explored and optimized. The SiNW-FET biosensor has good sensitivity with a detection limit of 0.01 fg/mL toward protein. The current change value shows a linear upward trend with the increase in protein concentration in the range of 1 fg/mL to 100 µg/mL. One whole test cycle can be accomplished within only 30 s. More importantly, a good distinction was realized in the sputum without pretreatment between normal people and TB patients, which greatly shortened the TB detection time (only 2-5 min, considering the dilution of sputum). Compared with other methods, the SiNW-FET biosensor can detect MTB with a remarkably broad dynamic linear range in a shorter time.

Advances in airborne microorganisms detection using biosensors: A critical review.

Humanity has been facing the threat of a variety of infectious diseases. Airborne microorganisms can cause airborne infectious diseases, which spread rapidly and extensively, causing huge losses to human society on a global scale. In recent years, the detection technology for airborne microorganisms has developed rapidly; it can be roughly divided into biochemical, immune, and molecular technologies. However, these technologies still have some shortcomings; they are time-consuming and have low sensitivity and poor stability. Most of them need to be used in the ideal environment of a laboratory, which limits their applications. A biosensor is a device that converts biological signals into detectable signals. As an interdisciplinary field, biosensors have successfully introduced a variety of technologies for bio-detection. Given their fast analysis speed, high sensitivity, good portability, strong specificity, and low cost, biosensors have been widely used in environmental monitoring, medical research, food and agricultural safety, military medicine and other fields. In recent years, the performance of biosensors has greatly improved, becoming a promising technology for airborne microorganism detection. This review introduces the detection principle of biosensors from the three aspects of component identification, energy conversion principle, and signal amplification. It also summarizes its research and application in airborne microorganism detection. The new progress and future development trend of the biosensor detection of airborne microorganisms are analyzed.

Particle Self-Aligning, Focusing, and Electric Impedance Microcytometer Device for Label-Free Single Cell Morphology Discrimination and Yeast Budding Analysis.

Microfluidic electric impedance flow cytometry (IFC) devices have been applied in single cell analysis, such as cell counting, volume discrimination, cell viability, etc. A cell's shape provides specific information about cellular physiological and pathological conditions, especially in microorganisms such as yeast. In this study, the particle orientation focusing was theoretically analyzed and realized by hydrodynamics. The pulse width (passing time for the particles) of the conductance signal was used to discriminate particle shapes. Spherical and rod-shaped particles with similar volumes/lengths were differentiated by the IFC device, using the impedance pulse parameters of the events. Then, typical late-budding, early budding, and unbudded yeast cells were distinguished by the width, amplitude, and ratio of width to amplitude (R) of the impedance pulse. The pulse amplitude and the R combination gate for identifying the late-budding yeast was estimated through the statistic results. Using the gate, the late-budding rates under different conditions were calculated. Late-budding rates obtained using our method showed a high correlation (R2 = 0.83) with the manual cell counting result and represented the budding status of yeast cells under different conditions proficiently. Thus, the late-budding rate calculated using the above method can be used as a qualitative parameter to assess the reproductive performance of yeast and whether a yeast culturing environment is optimal. This IFC device and cell shape discrimination method is very simple and could be applied in the fermentation industry and other microorganisms' discrimination as a rapid analysis technique in the future.

Dynamic sessile micro-droplet evaporation monitored by electric impedance sensing.

Studies of liquid evaporation on a solid surface are useful for wettability phenomena-related research, and can be applied in a series of scientific and industrial areas. However, traditional methods are not easy to be intergrated into small size to monitor evaporation process of a micro-droplet. In this paper, a micro-electrode array was used to measure the impedance of an electrolyte droplet, indicating the dynamic process of evaporation. This method uses the relationship between concentration and conductivity of the water solution to dynamically monitor the evaporation process. The dynamic impedance results were compared to weight and imaging data of droplet evaporation and demonstrate high correlation coefficient of the earlier 90% part of the sodium chloride droplet evaporation process (R 2 = 0.99). Our study proved that the height of the droplet will affect the impedance sensing result, and the solution used for droplet evaporation can be expanded to mixture of strong electrolyte solution such as phosphate buffered solution. Then the "impedance imaging" of the array monitored the evaporating speed differences of different sites of a sessile droplet. As the electrode array can be integrated into small size, this method is compatible for many other experimental systems and can be further used for evaporation studies and corresponding application areas.

A review of impedance measurements of whole cells.

Impedance measurement of live biological cells is widely accepted as a label free, non-invasive and quantitative analytical method to assess cell status. This method is easy-to-use and flexible for device design and fabrication. In this review, three typical techniques for impedance measurement, i.e., electric cell-substrate impedance sensing, Impedance flow cytometry and electric impedance spectroscopy, are reviewed from the aspects of theory, to electrode design and fabrication, and applications. Benefiting from the integration of microelectronic and microfluidic techniques, impedance sensing methods have expanded their applications to nearly all aspects of biology, including living cell counting and analysis, cell biology research, cancer research, drug screening, and food and environmental safety monitoring. The integration with other techniques, the fabrication of devices for certain biological assays, and the development of point-of-need diagnosis devices is predicted to be future trend for impedance sensing techniques.

[Advances in researches on thorax mechanical modeling and simulation during cardiopulmonary resuscitation].

Cardiopulmonary resuscitation (CPR) is vital for cardio arrest victims; in this field, researches have been aiming at its mechanism, operation guidelines, new CPR machines and so on. This paper summarized the studies on the mechanical characteristics of the thorax under CPR, and on the simulation work of sternal-displacement relationship in CPR manikins. The data from modeling work showed that the thorax's sternal-displacement relationship could be graphically represented by a hysteresis curve. As the actual CPR manikins' mechanical structures of the thorax are too simplified, it is necessary to do the work of improvement.