An effective splitting-and-recombination micromixer with self-rotated contact surface for wide Reynolds number range applications

It is difficult to mix two liquids on a microfluidic chip because the small dimensions and velocities effectively prevent the turbulence. This paper describes two 2-layer PDMS passive micromixers based on the concept of splitting and recombining the flow that exploits a self-rotated contact surface to increase the concentration gradients to obtain fast and efficient mixing. The designed micromixers were simulated and the mixing performance was assessed. The mixers have shown excellent mixing efficiency over a wide range of Reynolds number. The mixers were reasonably fabricated by multilayer soft lithography, and the experimental measurements were performed to qualify the mixing performance of the realized mixer. The results show that the mixing efficiency for one realized mixer is from 91.8% to 87.7% when the Reynolds number increases from 0.3 to 60, while the corresponding value for another mixer is from 89.4% to 72.9%. It is rather interesting that the main mechanism for the rapid mixing is from diffusion to chaotic advection when the flow rate increases, but the mixing efficiency has not obvious decline. The smart geometry of the mixers with total length of 10.25 mm makes it possible to be integrated with many microfluidic devices for various applications in μ-TAS and Lab-on-a-chip systems.     

A “twisted” microfluidic mixer suitable for a wide range of flow rate applications

This paper proposes a new “twisted” 3D microfluidic mixer fabricated by a laser writing/microfabrication technique. Effective and efficient mixing using the twisted micromixers can be obtained by combining two general chaotic mixing mechanisms: splitting/recombining and chaotic advection. The lamination of mixer units provides the splitting and recombination mechanism when the quadrant of circles is arranged in a two-layered serial arrangement of mixing units. The overall 3D path of the microchannel introduces the advection. An experimental investigation using chemical solutions revealed that these novel 3D passive microfluidic mixers were stable and could be operated at a wide range of flow rates. This micromixer finds application in the manipulation of tiny volumes of liquids that are crucial in diagnostics. The mixing performance was evaluated by dye visualization, and using a pH test that determined the chemical reaction of the solutions. A comparison of the tornado-mixer with this twisted micromixer was made to evaluate the efficiency of mixing. The efficiency of mixing was calculated within the channel by acquiring intensities using ImageJ software. Results suggested that efficient mixing can be obtained when more than 3 units were consecutively placed. The geometry of the device, which has a length of 30 mm, enables the device to be integrated with micro total analysis systems and other lab-on-chip devices.     

Mixing enhancement in microfluidic channel with a constriction under periodic electro-osmotic flow

We present a new approach to enhance mixing in T-type micromixers by introducing a constriction in the microchannel under periodic electro-osmotic flow. Two sinusoidal ac electric fields with 180° phase difference and similar dc bias are applied at the two inlets. The out of phase ac electric field induces oscillation of fluid interface at the junction of the two inlet channels and the constriction. Due to the constriction introduced at the junction, fluids from these two inlets form alternative plugs at the constricted channel. These plugs of fluids radiate downstream from the constriction into the large channel and form alternate thin crescent-shaped layers of fluids. These crescent-shaped layers of fluids increase tremendously the contact surface area between the two streams of fluid and thus enhance significantly the mixing efficiency. Experimental results and mixing mechanism analysis show that amplitude and frequency of the ac electric field and the length of the constriction govern the mixing efficiency.     

Taxonomy of quantities

The adjectives “generic”, “subgeneric”, “specific”, and “individual” are proposed to be added to the term “quantity”, depending on the case, in order to diminish the ambiguity of this term and to classify the different types of quantities.     

Numerical modeling of DNA-chip hybridization with chaotic advection

We present numerical simulations of DNA-chip hybridization, both in the “static” and “dynamical” cases. In the static case, transport of free targets is limited by molecular diffusion; in the dynamical case, an efficient mixing is achieved by chaotic advection, with a periodic protocol using pumps in a rectangular chamber. This protocol has been shown to achieve rapid and homogeneous mixing. We suppose in our model that all free targets are identical; the chip has different spots on which the probes are fixed, also all identical, and complementary to the targets. The reaction model is an infinite sink potential of width d_h, i.e., a target is captured as soon as it comes close enough to a probe, at a distance lower than d_h. Our results prove that mixing with chaotic advection enables much more rapid hybridization than the static case. We show and explain why the potential width d_h does not play an important role in the final results, and we discuss the role of molecular diffusion. We also recover realistic reaction rates in the static case.     

Photoresponsive microvalve for remote actuation and flow control in microfluidic devices

Microvalves with different actuation methods offer great integrability and flexibility in operation of lab-on-chip devices. In this work, we demonstrate a hydrogel-based and optically controlled modular microvalve that can be easily integrated within a microfluidic device and actuated by an off-chip laser source. The microvalve is based on in-channel trapping of microgel particles, which are composed of poly(N-isopropylacrylamide) and polypyrrole nanoparticles. Upon irradiation by a near-infrared (NIR) laser, the microgel undergoes volumetric change and enables precisely localized fluid on/off switching. The response rate and the “open” duration of the microvalve can be simply controlled by adjusting the laser power and exposure time. We showed that the trapped microgel can be triggered to shrink sufficiently to open a channel within as low as ∼1–2 s; while the microgel swells to re-seal the channel within ∼6–8 s. This is so far one of the fastest optically controlled and hydrogel-based microvalves, thus permitting speedy fluidic switching applications. In this study, we successfully employed this technique to control fluidic interface between laminar flow streams within a Y-junction device. The optically triggered microvalve permits flexible and remote fluidic handling, and enables pulsatile in situ chemical treatment to cell culture in an automatic and programmed manner, which is exemplified by studies of chemotherapeutic drug induced cell apoptosis under different drug treatment strategies. We find that cisplatin induced apoptosis is significantly higher in cancer cells treated with a pulsed dose, as compared to continuous flow with a sustained dose. It is expected that our NIR-controlled valving strategy will provide a simple, versatile, and powerful alternative for liquid handling in microfluidic devices.     

History of the preanalytical phase: a personal view

In the 70ies of the last century, ther term “preanalytical phase” was introduced in the literature. This term describes all actions and aspects of the “brain to brain circle” of the medical laboratory diagnostic procedure happening before the analytical phase. The author describes his personal experiences in the early seventies and the following history of increasing awareness of this phase as the main cause of “laboratory errors”. This includes the definitions of influence and interference factors as well as the first publications in book, internet, CD-Rom and recent App form over the past 40 years. In addition, a short summary of previous developments as prerequesits of laboratory diagnostic actions is described from the middle age matula for urine collection to the blood collection tubes, anticoagulants and centrifuges. The short review gives a personal view on the possible causes of missing awareness of preanalytical causes of error and future aspects of new techniques in regulation of requests to introduction of quality assurance programs for preanalytical factors.     

Novel index for micromixing characterization and comparative analysis

The most basic micromixer is a T- or Y-mixer, where two confluent streams mix due to transverse diffusion. To enhance micromixing, various modifications of T-mixers are reported such as heterogeneously charged walls, grooves on the channel base, geometric variations by introducing physical constrictions, etc. The performance of these reported designs is evaluated against the T-mixer in terms of the deviation from perfectly mixed state and mixing length (device length required to achieve perfect mixing). Although many studies have noticed the reduced flow rates for improved mixer designs, the residence time is not taken into consideration for micromixing performance evaluation. In this work, we propose a novel index, based on residence time, for micromixing characterization and comparative analysis. For any given mixer, the proposed index identifies the nondiffusive mixing enhancement with respect to the T-mixer. Various micromixers are evaluated using the proposed index to demonstrate the usefulness of the index. It is also shown that physical constriction mixer types are equivalent to T-mixers. The proposed index is found to be insightful and could be used as a benchmark for comparing different mixing strategies.     

Survey on reporting of epithelial cells in urine sediment as part of external quality assessment programs in Brazilian laboratories

IntroductionEpithelial cells (ECs) are structures regularly observed during urine microscopy analysis. The correct identification of EC subtypes can be useful since renal tubular epithelial cells (RTECs) are clinically relevant. We investigate the urinary ECs report and the judgement of its clinical importance by Brazilian laboratories.Materials and methodsA survey with four questions was made available to participants of the Urinalysis External Quality Assessment Program (EQAP) from Controllab. Laboratories composed 3 groups: (1) differentiating ECs subtypes: “squamous”, “transitional” and “RTECs”; (2) differentiating ECs subtypes: “squamous” or “non-squamous” cells; (3) without ECs subtype identification. Participants did not necessarily answer to all questions and the answers were evaluated both within the same laboratory’s category and within different categories of laboratories.ResultsA total of 1336 (94%) laboratories answered the survey; Group 1, 119/140 (85%) reported that ECs differentiation is important to the physician and 62% want to be evaluated by EQAP, while in Group 3, 455/1110 (41%) reported it is useful to them, however only 25% want be evaluated by EQAP. Group 2 laboratories 37/51 (73%) reported that the information is important, but only 13/52 (25%) are interested in an EQAP with differentiation of the 3 ECs subtypes.ConclusionMost of the laboratories do not differentiate ECs in the three subtypes, despite the clinical importance of RTECs. Education of laboratory staff about the clinical significance of urinary particles should be considered a key priority.     

Mixing enhancement in T-junction microchannel with acoustic streaming induced by triangular structure

The T-shaped microchannel system is used to mix similar or different fluids, and the laminar flow nature makes the mixing at the entrance junction region a challenging task. Acoustic streaming is a steady vortical flow phenomenon that can be produced in the microchannel by oscillating acoustic transducer around the sharp edge tip structure. In this study, the acoustic streaming is produced using a triangular structure with tip angles of 22.62°, 33.4°, and 61.91°, which is placed at the entrance junction region and mixes the inlets flow from two directions. The acoustic streaming flow patterns were investigated using micro-particle image velocimetry (μPIV) in various tip edge angles, flow rate, oscillation frequency, and amplitude. The velocity and vorticity profiles show that a pair of counter-rotating streaming vortices were created around the sharp triangle structure and raised the Z vorticity up to 10 times more than the case without acoustic streaming. The mixing experiments were performed by using fluorescent green dye solution and de-ionized water and evaluated its performance with the degree of mixing (M) at different amplitudes, flow rates, frequencies, and tip edge angles using the grayscale value of pixel intensity. The degree of mixing characterized was found significantly improved to 0.769 with acoustic streaming from 0.4017 without acoustic streaming, in the case of 0.008 μl/min flow rate and 38 V oscillation amplitude at y = 2.15 mm. The results suggested that the creation of acoustic streaming around the entrance junction region promotes the mixing of two fluids inside the microchannel, which is restricted by the laminar flow conditions.     

Failure to review STAT clinical laboratory requests and its economical impact

Background

Failure to follow-up laboratory test results has been described as one of the major processes contributing to unsafe patient care. Currently, most of the laboratories do not know with certainty not only their rate of missed (or unreviewed) requests but the economical cost and impact that this issue implies. The aim of our study was to measure that rate and calculate the resulting costs.

Material and methods

In January 2015, we checked in our Laboratory Information Management System (LIMS) for every emergency request from 1^st July 2011 to 30^th June 2014, if they had been reviewed by any allowed user or not. 319,064 requests were ordered during that period of time. Results were expressed as “ordered requests”, “missed requests” and its percentage. Additionally, total cost of missed requests was calculated in euros (€). “Non-productive days” were theorised (as the days producing requests that were not reviewed) based on these results.

Results

7924 requests (2.5%) were never reviewed by clinicians. This represented a total cost of 203,039 € and 27 “non-productive” days in three years. Significant differences between inpatients, outpatients and emergency department as well as different emergencies units were found after application of statistical analysis.

Conclusions

In terms of resources, never reviewed or missed requests appear to be a not negligible problem for the clinical laboratory management. Electronic result delivery, with electronic endorsement to indicate follow-up of requests along with better systems of electronic requesting should be investigated as a way of improving patient outcomes and save unnecessary expenses.Key words: quality indicators, health care, extra-analytical phase, total quality management, clinical laboratory information systems     

A simplified design of the staggered herringbone micromixer for practical applications

We demonstrated a simple method for the device design of a staggered herringbone micromixer (SHM) using numerical simulation. By correlating the simulated concentrations with channel length, we obtained a series of concentration versus channel length profiles, and used mixing completion length L_m as the only parameter to evaluate the performance of device structure on mixing. Fluorescence quenching experiments were subsequently conducted to verify the optimized SHM structure for a specific application. Good agreement was found between the optimization and the experimental data. Since L_m is straightforward, easily defined and calculated parameter for characterization of mixing performance, this method for designing micromixers is simple and effective for practical applications.     

基于结构方程的居民生活垃圾分类行为研究

居民是城市生活垃圾分类回收系统中的关键主体之一,其分类回收参与率的高低直接关系到整个城市垃圾分类工作的成败。为分析居民垃圾分类行为决策机制,本文以苏州市为案例,对其中心城区居民开展实地问卷调查,并运用结构方程统计分析方法,探讨影响居民生活垃圾分类行为的主要因素和作用路径。研究结果表明：居民生活垃圾分类行为主要由4个主观因素和7个外部情境因素共同作用的结果,且外部条件因素的综合影响是个体主观因素的近2倍,其中,“环境设施和服务”潜变量相关因素对居民行为的综合影响最大,公共宣传教育、回收设施便利性、分类设施便利性、参与意愿和环保意识为对居民垃圾生活处理行为影响最显著的前5个因素。为进一步加快推进中国城市生活垃圾分类工作,建议加强城市生活垃圾分类回收配套设施规划建设,广泛开展垃圾分类回收宣传教育,完善相关法律法规并建立有效的激励约束机制。     

Influence of buoyancy-driven flow on mass transfer in a two-stream microfluidic channel: Introduction of cryoprotective agents into cell suspensions

A variety of methods have been used to introduce chemicals into a stream or to mix two or more streams of different compositions using microfluidic devices. In the following paper, the introduction of cryoprotective agents (CPAs) used during cryopreservation of cells in order to protect them from freezing injuries and increase viability post thaw is described. Dimethylsulphoxide (DMSO) is the most commonly used CPA. We aim to optimize the operating conditions of a two-stream microfluidic device to introduce a 10% vol/vol solution of DMSO into a cell suspension. Transport behavior of DMSO between two streams in the device has been experimentally characterized for a spectrum of flow conditions (0.7 < Re < 10), varying initial donor stream concentrations, (1% vol/vol < C_o < 15% vol/vol) and different flow rate fractions (0.23 < f_q < 0.77). The outlet cell stream concentration is analyzed for two different flow configurations: one with the cell stream flowing on top of the DMSO-rich donor stream, and the other with the cell stream flowing beneath the heavy DMSO-laden stream. We establish a transition from a diffusive mode of mass transfer to gravity-influenced convective currents for Atwood numbers (A_t) in the range of (1.7 × 10⁻³ < A_t < 3.1 × 10⁻³) for the latter configuration. Flow visualization with cells further our understanding of the effect of A_t on the nature of mass transport. Cell motion studies performed with Jurkat cells confirm a high cell recovery from the device while underscoring the need to collect both the streams at the outlet of the device and suggesting flow conditions that will help us achieve the target DMSO outlet concentration for clinical scale flow rates of the cell suspension.     

Evaluation of preanalytical and postanalytical phases in clinical biochemistry laboratory according to IFCC laboratory errors and patient safety specifications

IntroductionThe aim of the study was to determine the current state of laboratory’s extra-analytical phase performance by calculating preanalytical and postanalytical phase quality indicators (QIs) and sigma values and to compare obtained data according to desired quality specifications and sigma values reported by The International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) Working Group – Laboratory errors and Patient Safety.Materials and methodsPreanalytical and postanalytical phase data were obtained through laboratory information system. Rejected samples in preanalytical phase were grouped according to reasons for rejection and frequencies were calculated both monthly and for 2019. Sigma values were calculated according to “short term sigma” table.ResultsThe number of rejected samples in laboratory was 643 out of 191,831 in 2019. Total preanalytical phase rejection frequency was 0.22%. According to the reasons for rejection, QIs and sigma values were: “Samples with excessive transportation time”: 0.0036 and 5.47; “Samples collected in wrong container” 0.02 and 5.11. In December, QIs and sigma values were: “Samples with excessive transportation time”: 0.01 and 5.34; “Samples collected in wrong container”: 0.03 and 4.98. The postanalytical QIs and sigma values were: “Reports delivered outside the specified time”: 0.34 and 4.21; “Turn around time of potassium”: 56 minute and 3.84, respectively. There were no errors in “Critical values of inpatients and outpatients notified after a consensually agreed time”.ConclusionsExtra-analytical phase was evaluated by comparing it with the latest quality specifications and sigma values which will contribute to improving the quality of laboratory medicine.     

Decision support system through automatic algorithms and electronic request in diagnosis of anaemia for primary care patients

IntroductionAn appropriate management of anaemia laboratory tests is crucial for a correct diagnosis and treatment. A non-sequential “shotgun” approach (where every anaemia related test is ordered) causes workload and cost increases and could be potentially harmful. We have implemented a Decision Support System through our laboratory information system (LIMS) based on reflexive algorithms and automatic generation of interpretative reports specifically in diagnosis of anaemia for primary care patients.Materials and methodsWhen a request contained an “Anaemia Suspicion Study” profile, more than twenty automatic reflexive rules were activated in our LIMS based upon laboratory results. These rules normally involved the addition of reflexive tests. A final report was automatically generated for each interpretation which was always reviewed for their validity by two staff pathologists. We measured the impact of this system in the ordering of most common anaemia related tests and if a proper treatment was established based on the interpretive report.ResultsFrom all the studies performed, only 12% were positive being “iron deficiency” and “anaemia of chronic disease” the most frequent causes, 62% and 17%, respectively. Proper treatment was established in 88% of these anaemic patients. Total iron, transferrin, ferritin, folate and vitamin B12 demand decreased substantially after implementation representing a cost reduction of 40% only for these five tests.ConclusionsOur system has easily improved patient outcomes, advising on individual clinical cases. We have also noticeably reduced the number of over-requested tests and laboratory costs.     

An efficient micromixer based on multidirectional vortices due to baffles and channel curvature

An efficient planar micromixer based on multidirectional vortices in a curved channel with radial baffles is proposed and examined in this work. The curvature of the microchannel and the radial baffles induce vortices in different directions. The multidirectional vortices and the converging-diverging flow caused by the baffles contribute together to the enhancement of mixing. The micromixer is fabricated with polydimethylsiloxane by a single planar microlithography process and the mixing behaviors are observed by a confocal spectral microscope imaging system to validate the simulation obtained by a commercial code. The simulation and experimental results are in reasonable agreement. The concentration distributions and flow patterns obtained reveal the following trends. (i) The mixing efficiency of the basic C-shaped micromixer with the first baffle attached to the internal cylinder and the second attached to the external cylinder is better than that of the C-shaped micromixer with inverted arrangement of baffles. (ii) When the radius of the curved channel and the width of the passage between the baffle and the cylindrical wall are small enough and the Reynolds number (Re) is large enough, an extra separation vortex develops in the downstream of the second baffle. This phenomenon is one of the reasons of trend (i). (iii) A micromixer consisting of a few basic C-shaped micromixers connected by straight channels may generate a high degree of mixing for the case with a large Re.     

An integrated microfluidic platform to fabricate single-micrometer asymmetric giant unilamellar vesicles (GUVs) using dielectrophoretic separation of microemulsions

We present a microfluidic technique that generates asymmetric giant unilamellar vesicles (GUVs) in the size range of 2–14 μm. In our method, we (i) create water-in-oil emulsions as the precursors to build synthetic vesicles, (ii) deflect the emulsions across two oil streams containing different phospholipids at high throughput to establish an asymmetric architecture in the lipid bilayer membranes, and (iii) direct the water-in-oil emulsions across the oil–water interface of an oscillating oil jet in a co-flowing confined geometry to encapsulate the inner aqueous phase inside a lipid bilayer and complete the fabrication of GUVs. In the first step, we utilize a flow-focusing geometry with precisely controlled pneumatic pressures to form monodisperse water-in-oil emulsions. We observed different regimes in forming water-in-oil multiphase flows by changing the applied pressures and discovered a hysteretic behavior in jet breakup and droplet generation. In the second step of GUV fabrication, an oil stream containing phospholipids carries the emulsions into a separation region where we steer the emulsions across two parallel oil streams using active dielectrophoretic and pinched-flow fractionation separations. We explore the effect of applied DC voltage magnitude and carrier oil stream flow rate on the separation efficiency. We develop an image processing code that measures the degree of mixing between the two oil streams as the water-in-oil emulsions travel across them under dielectrophoretic steering to find the ideal operational conditions. Finally, we utilize an oscillating co-flowing jet to complete the formation of asymmetric giant unilamellar vesicles and transfer them to an aqueous phase. We investigate the effect of flow rates on properties of the co-flowing jet oscillating in the whipping mode (i.e., wavelength and amplitude) and define the phase diagram for the oil-in-water jet. Assays used to probe the lipid bilayer membrane of fabricated GUVs showed that membranes were unilamellar, minimal residual oil remained trapped between the two lipid leaflets, and 83% asymmetry was achieved across the lipid bilayers of GUVs.     

The pits and falls of graphical presentation

Graphics are powerful tools to communicate research results and to gain information from data. However, researchers should be careful when deciding which data to plot and the type of graphic to use, as well as other details. The consequence of bad decisions in these features varies from making research results unclear to distortions of these results, through the creation of “chartjunk” with useless information. This paper is not another tutorial about “good graphics” and “bad graphics”. Instead, it presents guidelines for graphic presentation of research results and some uncommon, but useful examples to communicate basic and complex data types, especially multivariate model results, which are commonly presented only by tables. By the end, there are no answers here, just ideas meant to inspire others on how to create their own graphics.     

Surface tension effects on submerged electrosprays

Electrosprays are a powerful technique to generate charged micro/nanodroplets. In the last century, the technique has been extensively studied, developed, and recognized with a shared Nobel price in Chemistry in 2002 for its wide spread application in mass spectrometry. However, nowadays techniques based on microfluidic devices are competing to be the next generation in atomization techniques. Therefore, an interesting development would be to integrate the electrospray technique into a microfluidic liquid-liquid device. Several works in the literature have attempted to build a microfluidic electrospray with disputable results. The main problem for its integration is the lack of knowledge of the working parameters of the liquid-liquid electrospray. The “submerged electrosprays” share similar properties as their counterparts in air. However, in the microfluidic generation of micro/nanodroplets, the liquid-liquid interfaces are normally stabilized with surface active agents, which might have critical effects on the electrospray behavior. In this work, we review the main properties of the submerged electrosprays in liquid baths with no surfactant, and we methodically study the behavior of the system for increasing surfactant concentrations. The different regimes found are then analyzed and compared with both classical and more recent experimental, theoretical and numerical studies. A very rich phenomenology is found when the surface tension is allowed to vary in the system. More concretely, the lower states of electrification achieved with the reduced surface tension regimes might be of interest in biological or biomedical applications in which excessive electrification can be hazardous for the encapsulated entities.