Filtration Of Microparticles Research Articles

Dispersion-free inertial focusing (DIF) for high-yield polydisperse micro-particle filtration and analysis.

Inertial focusing excels at the precise spatial ordering and separation of microparticles by size within fluid flows. However, this advantage, resulting from its inherent size-dependent dispersion, could turn into a drawback that challenges applications requiring consistent and uniform positioning of polydisperse particles, such as microfiltration and flow cytometry. To overcome this fundamental challenge, we introduce Dispersion-Free Inertial Focusing (DIF). This new method minimizes particle size-dependent dispersion while maintaining the high throughput and precision of standard inertial focusing, even in a highly polydisperse scenario. We demonstrate a rule-of-thumb principle to reinvent an inertial focusing system and achieve an efficient focusing of particles ranging from 6 to 30 μm in diameter onto a single plane with less than 3 μm variance and over 95% focusing efficiency at highly scalable throughput (2.4-30 mL h-1) - a stark contrast to existing technologies that struggle with polydispersity. We demonstrated that DIF could be applied in a broad range of applications, particularly enabling high-yield continuous microparticle filtration and large-scale high-resolution single-cell morphological analysis of heterogeneous cell populations. This new technique is also readily compatible with the existing inertial microfluidic design and thus could unleash more diverse systems and applications.

From the teapot effect to tap-triggered self-wetting: a 3D self-driving sieve for whole blood filtration

Achieving passive microparticle filtration with micropore membranes is challenging due to the capillary pinning effect of the membranes. Inspired by the teapot effect that occurs when liquid (tea) is poured from a teapot spout, we proposed a tap-triggered self-wetting strategy and utilized the method with a 3D sieve to filter rare cells. First, a 3D-printed polymer tap-trigger microstructure was implemented. As a result, the 3 µm micropore membrane gating threshold (the pressure needed to open the micropores) was lowered from above 3000 to 80 Pa by the tap-trigger microstructure that facilated the liquid leakage and spreading to self-wet more membrane area in a positive feedback loop. Then, we implemented a 3D cone-shaped cell sieve with tap-trigger microstructures. Driven by gravity, the sieve performed at a high throughput above 20 mL/min (DPBS), while the micropore size and porosity were 3 µm and 14.1%, respectively. We further filtered leukocytes from whole blood samples with the proposed new 3D sieve, and the method was compared with the traditional method of leukocyte isolation by chemically removing red blood cells. The device exhibited comparable leukocyte purity but a higher platelet removal rate and lower leukocyte simulation level, facilitating downstream single-cell analysis. The key results indicated that the tap-triggered self-wetting strategy could significantly improve the performance of passive microparticle filtration.

Radially actuating conducting polymer microactuators as gates for dynamic microparticle sieve based on printed microfluidics

A new radially expanding conducting polymer microactuator is presented. The radially expanding microactuators are used as electroactive gates in an electrically controlled microparticle sieve. A novel configuration to dynamically filter particles of different sizes in a microfluidic chip is conceptualized. Micropillars of SU-8 combined with conducting polymers to provide the radial actuation are positioned in a microfluidic chip with a specifically designed 3D printed housing to allow for selective filtration of microparticles with varied sizes. These pillar-shaped microactuators of polypyrrole actuate radially to function as dynamic gates for the fluidic channel, controlling the porosity of the filter allowing for the filtration of specific size of microparticles. This sieve design provides user defined channel width modulation with external stimuli. Photolithography and electrochemical polymerizations are combined with additive manufacturing to fabricate the individual functional parts of the microfluidic filter. To demonstrate the new conceptual filter design, we have shown filtration of microparticles of the sizes 60, 80, 90 and 100 µm by electrically actuating micropillars of the dynamic gate. The flow and aggregation of the microparticles were analysed at the dynamic gates to characterize the performance of the filter.

Inertial focusing and filtration of microparticles with expansion–contraction structures in microchannel

Inertial focusing and filtration of microparticles with expansion–contraction structures in microchannel

Synthesis of Mesoporous Carbon from PVDF and PTFE via Defluorination of 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU)

Porous carbon has found commercial applications as a filter material based on the sorption ability of its pores. The pore size and surface properties of the carbon can be varied depending on the type of particles to be filtered. Here, mesoporous carbon was induced through the pyrolysis of polyvinylidene fluoride (PVDF) to fabricate a porous material for microparticle filtration. Because removal of the constituent fluorine at elevated temperature leaves small-sized micropores, the PVDF precursor mainly generates micropores during pyrolysis. To suppress the micropore evolution mechanism, the PVDF precursor was defluorinated before the heat treatment using 1,8-Diazabicyclo[5.4.0]undec-7-ene(DBU) and then pyrolyzed. The suppressed evolution of the micropores during carbon synthesis leads to a lower specific surface area, suggesting low adsorption capacity. The polytetrafluoroethylene (PTFE) was mixed with the PVDF precursor to induce mesoporosity. The PVDF precursor mixed with the PTFE enhanced the surface area since the PTFE could be removed, leaving mesopores after pyrolysis. The effect of the defluorination process on the porosity was investigated by varying the ratio of DBU to vinylidene fluoride unit (1, 5, 10, 20) in the precursor solution. With higher DBU content in the precursor, the micropore evolution was reduced with a lower specific surface area. The porous carbons synthesized from the precursor with a high DBU amount (DBU/vinylidene fluoride unit = 5, 10, 20) were almost entirely composed of mesopores. In addition, the higher DBU content reduced the hydrophilicity of the synthesized carbon. In summary, to separate and absorb relatively large impurities, the mesoporous carbon should be synthesized using a mixture of PVDF and PTFE precursor with an appropriate amount of DBU for a higher specific surface area.

A bioinspired, passive microfluidic lobe filtration system.

Size-based microfluidic filtration systems can be affected by clogging, which prevents their use in high-throughput and continuous applications. To address these concerns, we have developed two microfluidic lobe filters bioinspired by the filtration mechanism of two species of manta ray. These chips enable filtration of particles around 10-30 μm with precise control and high throughput by using two arrays of equally spaced filter lobes. For each filter design, we investigated multiple inlet flow rates and particle sizes to identify successful operational parameters. Filtration efficiency increases with fluid flow rate, suggesting that particle inertial effects play a key role in lobe filter separation. Microparticle filtration efficiencies up to 99% were obtainable with inlet flow rates of 20 mL min-1. Each filter design successfully increased microparticle concentrations by a factor of two or greater at different inlet flow rates ranging from 6-16 mL min-1. At higher inlet flow rates, ANSYS Fluent simulations of each device revealed a complex velocity profile that contains three local maxima and two inflection points. Ultimately, we show that distances from the lobe array to the closest local maxima and inflection point of the velocity profile can be used to successfully estimate lobe filtration efficiency at each operational flow rate.

Microparticle filtration ability of pervious concrete mixed with recycled synthetic fibers

In this study, the properties of fiber reinforced pervious concrete (FRPC) was investigated, with the focus on the ability to filter microparticles. The fibers were obtained from recycle plastic bags and mixed with pervious concrete at the rate of 0 to 0.3% by volume fractions. In addition, the effect of silica fume and aggregate size was also investigated. The experimental part consisted of tests on mechanical properties (i.e., compressive and flexural strength) and physical properties (i.e., void ratio test, permeability, and microparticle filtration). Two types of microparticles were used: microplastics (MPs) and microrubbers (MRs). Results indicated that the addition of fibers led to the decrease in both compressive and flexural strengths. In the case of physical properties, fibers provided direct effect to the void system of concrete, which led to the reduction of permeability and void ratio as well as the improvement in microparticle filtration ability. The addition of silica fume and the use of smaller size aggregates also gave positive effects on the microparticle filtration ability of pervious concrete.

Water tracking in surface water, groundwater and soils using free and alginate-chitosan encapsulated synthetic DNA tracers

Investigating contamination pathways and hydraulic connections in complex hydrological systems will benefit greatly from multi-tracer approaches. The use of non-toxic synthetic DNA tracers is promising, because unlimited numbers of tracers, each with a unique DNA identifier, could be used concurrently and detected at extremely low concentrations. This study aimed to develop multiple synthetic DNA tracers as free molecules and encapsulated within microparticles of biocompatible and biodegradable alginate and chitosan, and to validate their field utility in different systems. Experiments encompassing a wide range of conditions and flow rates (19cm/day-39km/day) were conducted in a stream, an alluvial gravel aquifer, a fine coastal sand aquifer, and in lysimeters containing undisturbed silt loam over gravels. The DNA tracers were identifiable in all field conditions investigated, and they were directly detectable in the stream at a distance of at least 1km. The DNA tracers showed promise at tracking fast-flowing water in the stream, gravel aquifer and permeable soils, but were unsatisfactory at tracking slow-moving groundwater in the fine sand aquifer. In the surface water experiments, the microencapsulated DNA tracers' concentrations and mass recoveries were 1-3 orders of magnitude greater than those of the free DNA tracers, because encapsulation protected them from environmental stressors and they were more negatively charged. The opposite was observed in the gravel aquifer, probably due to microparticle filtration by the aquifer media. Although these new DNA tracers showed promise in proof-of-concept field validations, further work is needed before they can be used for large-scale investigations.

Self-detached membranes with well-defined pore size, shape and distribution fabricated by underexposure photolithography

An underexposure photolithography method was developed to fabricate self-detached polymer micropore membranes with uniform pore size, shape, and arrangement. The key to this technique is to control and adjust the gradient of exposure dose projected into the film of photoresist. This new approach abandons sacrificial layers used in previous techniques. Negative SU-8 was chosen as an example photoresist to demonstrate its feasibility. Membranes with specially tailored sizes and shapes of micropores could be produced on diverse substrates. The coefficient of variation of pore size was only 1%, much lower than that for conventional microporous membranes. Moreover, due to self-detaching without a sacrificial layer, the membranes were flat and free of residual stress and deformation. This novel photolithographic approach opened a new avenue to manufacture high-quality membranes that could broaden the applications of microparticle filtration, separation, and sorting.

MicroCT analysis of connectivity in porous structures: optimizing data acquisition and analytical methods in the context of tissue engineering.

Micro-computed X-ray tomography (MicroCT) is one of the most powerful techniques available for the three-dimensional characterization of complex multi-phase or porous microarchitectures. The imaging and analysis of porous networks are of particular interest in tissue engineering due to the ability to predict various large-scale cellular phenomena through the micro-scale characterization of the structure. However, optimizing the parameters for MicroCT data capture and analyses requires a careful balance of feature resolution and computational constraints while ensuring that a structurally representative section is imaged and analysed. In this work, artificial datasets were used to evaluate the validity of current analytical methods by considering the effect of noise and pixel size arising from the data capture, and intrinsic structural anisotropy and heterogeneity. A novel ‘segmented percolation method’ was developed to exclude the effect of anomalous, non-representative features within the datasets, allowing for scale-invariant structural parameters to be obtained consistently and without manual intervention for the first time. Finally, an in-depth assessment of the imaging and analytical procedures are presented by considering percolation events such as micro-particle filtration and cell sieving within the context of tissue engineering. Along with the novel guidelines established for general pixel size selection for MicroCT, we also report our determination of 3 μm as the definitive pixel size for use in analysing connectivity for tissue engineering applications.

Structurally graduated collagen scaffolds applied to the ex vivo generation of platelets from human pluripotent stem cell-derived megakaryocytes: Enhancing production and purity

Platelet transfusions are a key treatment option for a range of life threatening conditions including cancer, chemotherapy and surgery. Efficient ex vivo systems to generate donor independent platelets in clinically relevant numbers could provide a useful substitute. Large quantities of megakaryocytes (MKs) can be produced from human pluripotent stem cells, but in 2D culture the ratio of platelets harvested from MK cells has been limited and restricts production rate. The development of biomaterial cell supports that replicate vital hematopoietic micro-environment cues are one strategy that may increase in vitro platelet production rates from iPS derived Megakaryocyte cells.In this paper, we present the results obtained generating, simulating and using a novel structurally-graded collagen scaffold within a flow bioreactor system seeded with programmed stem cells. Theoretical analysis of porosity using micro-computed tomography analysis and synthetic micro-particle filtration provided a predictive tool to tailor cell distribution throughout the material. When used with MK programmed stem cells the graded scaffolds influenced cell location while maintaining the ability to continuously release metabolically active CD41 + CD42 + functional platelets. This scaffold design and novel fabrication technique offers a significant advance in understanding the influence of scaffold architectures on cell seeding, retention and platelet production.

Viscosity and surface tension in the biological microparticle filtration process

The paper uses a simple model to present the effects of viscosity and surface tension on the basic parameters of the biological microparticle filtration process. A single filtration membrane with a uniform transmission structure in 2D space was used for the purposes of this analysis. The filtration system relations are described for the threshold filtration force, maximum and minimum height of the filtered liquid to guarantee stable filtration in a gravity field, and the duration of the process and forces affecting the microparticles during filtration. Based on these relations, the summary shows sample calculations for the base model of negative filtration of biological particles. Keywords: microparticle filtration, filtration screens, biological particle filtration, filtration membranes, sedimentation, kinematic viscosity, dynamic viscosity, liquid surface tension

Filtration of micro-particles within multi-fiber arrays by adhesive DEM-CFD simulation

A 3D multi-time scale discrete element method-computational fluid dynamic (DEM-CFD) coupling approach was applied to investigate the filtration of micron-sized particles by different types of fiber arrays. Both the pressure drop and the filtration efficiency were examined to indicate the filtration performance of the fiber arrays. Fibers that were uniformly arrayed in a parallel or staggered manner were compared. Results showed that the staggered array showed a better performance than the parallel array in terms of both pressure drop and filtration efficiency. Further, we compared the performance of different staggered arrays, i.e. a regular case, one densified in the front layers and another densified in the back layers. The front densified array was found to enter the clogging and cake filtration stage in the shortest time, leading to the highest filtration efficiency, but the highest pressure drop. The back densified array still achieved a much higher filtration efficiency, despite a much lower pressure drop comparable to that of the regular array. The results suggest that the two kinds of densified arrays may be suited for different purposes, e.g. baghouse filters or breathing masks.

Filtration of micro particles by laser-induced thermophoresis

Filtration of micro particles by laser-induced thermophoresis

Three Months of Human Haptoglobin Treatment Decreases Iron Deposition in the Kidneys of Townes Sickle Mice

Introduction: Haptoglobin (Hp), the scavenger for hemoglobin, and hemopexin (Hx), the scavenger for heme, are depleted in most patients with sickle cell disease due to chronic hemolysis. There is mounting evidence of the crucial role of free hemoglobin and/or free heme in mediating inflammatory and oxidative damage in sickle cell disease, including vaso-occlusion and acute chest syndrome. Purified Hp has been used in Japan for a variety of hemolytic conditions and has been proposed as a potential treatment for sickle cell disease. Although infusions of Hp or Hx have been shown to ameliorate vaso-occlusion, acute lung injury, and heme toxicity in sickle cell mouse models, no prior studies have examined the utility of chronic Hp treatment for amelioration of organ damage. We therefore studied the effect of 3 months of chronic Hp treatment in the Townes sickle mouse model.Methods: Male and female Townes mice (Stock number 013071, The Jackson Laboratory) were used for all experiments, starting at 1 or 3 months of age. SS genotype was confirmed by PCR and HPLC. Organ damage in the spleen, liver, and kidneys as previously described was confirmed. Human Hp solution was a kind gift from Bio Product Laboratory (BPL, Hertfordshire, UK). Hp or equivalent volume PBS control was administered intraperitoneally (IP) in the first cohort of 5 mice and then subcutaneously (SC) in the next two cohorts of 7 and 12 mice on a 48-72 hr dosing schedule of Monday, Wednesday, Friday for a period of 3 months. At the end of 3 months treatment, mice were evaluated by the following studies (with concurrent blinding to treatment group for most studies): plasma Hp (ELISA), plasma heme (QuantiChrom heme assay), urine osmolality (osmometer), urine albumin (ELISA), CBC (Advia 120), WBC differential (Advia 120 and manual count), red blood cell ektacyometry (ektacyometer), organ mass (percent of body weight), and organ histology.Results: Mouse Hp levels in SS Townes mice were confirmed to be markedly low compared to Townes AA mice (mean ± SD: SS 2 ± 1 versus AA 39 ± 4 ug/mL). Dose-finding experiments determined that a dose of 200-400 mg/kg IP or SC in SS mice resulted in a 24 hr peak concentration that was 5-14X supraphysiologic, variably physiologic at 48 hr, and absent or almost absent at 72 hr. Chronic dosing at the 400 mg/kg IP in SA mice showed no CBC or organ toxicity. Three successive cohorts of SS mice were treated with Hp (or equivalent volume of PBS): 200 mg/kg IP in 3-month old mice, 400 mg/kg SC in 3-month old mice, and 400 mg/kg SC in 1-month old mice. At the 400 mg/kg dosing levels, there was a significant decrease in iron deposition in the kidneys of both 4-month and 6-month old mice (treatment started at 1-month and 3 months, respectively) (Table 1). There was also a trend towards decreased liver infarction in 6-month old mice (Table 2).Discussion: Functional binding of the administered human Hp to the human Hb of the Townes mice likely occurred, as evidenced by the decrease in iron deposition in the kidneys, suggesting that formation of the complex prevents filtration of Hb into the kidneys. Surprisingly, kidney function as measured by urine osmolality or albumin excretion was not improved, which may be explained by continued heme-laden red cell microparticle filtration (Camus SM, Blood 2015). Encouragingly, however, a trend towards decreased liver infarction in older mice was observed. The less-than-expected effect of Hp on mouse disease severity may also be explained by: 1) continuous physiologic Hp concentrations not being maintained with the dosing frequency while continued hemolysis releases Hb every minute of the day, and 2) CD163-mediated uptake in mice seems to only account for a part of the Hb clearance as opposed to in humans (Etzerodt A, Antioxid Redox Signal 2013). Despite the limitations of the SCD mouse model, the current study suggests haptogobin infusions could be beneficial in SCD patients.Acknowledgment: The authors are grateful to Sandra Suzuka for performing the HPLC.Table 1Table 1. Kidney iron deposition (scale 1-10)Treatment group6-month old SS4-month old SS400 mg/kg Hp4.0 ± 1.41 ± 1.1PBS9.3 ± 0.65 ± 2.9p-value0.0020.02Table 2Liver infarction (scale 1-10)Treatment group6-month old SS4-month old SS400 mg/kg Hp2.6 ± 2.03.7 ± 2.8PBS6.3 ± 2.43.8 ± 2.3p-value0.070.91 DisclosuresBelcher:Biogen Idec: Research Funding; Seattle Genetics: Research Funding; CSL Behring: Research Funding. Vercellotti:CSL Behring: Research Funding; Seattle Genetics: Research Funding; Cydan: Research Funding; Biogen Idec: Research Funding.

Modulation of rotation-induced lift force for cell filtration in a low aspect ratio microchannel.

Cell filtration is a critical step in sample preparation in many bioapplications. Herein, we report on a simple, filter-free, microfluidic platform based on hydrodynamic inertial migration. Our approach builds on the concept of two-stage inertial migration which permits precise prediction of microparticle position within the microchannel. Our design manipulates equilibrium positions of larger microparticles by modulating rotation-induced lift force in a low aspect ratio microchannel. Here, we demonstrate filtration of microparticles with extreme efficiency (>99%). Using multiple prostate cell lines (LNCaP and human prostate epithelial tumor cells), we show filtration from spiked blood, with 3-fold concentration and >83% viability. Results of a proliferation assay show normal cell division and suggest no negative effects on intrinsic properties. Considering the planar low-aspect-ratio structure and predictable focusing, we envision promising applications and easy integration with existing lab-on-a-chip systems.

Ultrasonic flow-through filtration of microparticles in a microfluidic channel using frequency sweep technique

Many studies have been conducted on the filtration of microparticles using the acoustic radiation force of ultrasonic standing wave. The present work concerns a flow-through particle filtration method by utilizing frequency varying ultrasound. The periodical frequency sweep of the ultrasonic standing wave translocates particles across a microchannel, where particles in fluid flow are filtrated without barriers. The present filtration technique in a microfluidic channel was proposed conceptually in the 1990s. However, its experimental realization on actual particles in a microfluidic channel has not been carried out in a notable way. Several sizes of polystyrene microspheres (10 μm to 90 μm) and silicon carbide (SiC) particles (37 μm) suspended in water were applied as a test sample. For filtration of those particles, a Y-branched microfluidic channel with one inlet and two outlets was made out of steel and acrylic as a form of modulized device. Ultrasound of a few MHz in band frequency (1.75 MHz to 3.05 MHz) was transmitted into one side of the channel wall to generate a standing wave field in fluid flow. The periodical frequency sweep operation showed successful filtration performance, whereby particles in water flowed into one outlet and purified water flowed into the other outlet of the Y branch of the channel.

PWE-294 Microparticle dependent procoagulant activity and thrombin generation is increased in patients with cirrhosis induced coagulopathy

IntroductionRecent data suggests stable cirrhotics may have a hypercoagulable phenotype. Microparticles (MPs) are submicron plasma particles formed by the exocytic budding of cell membranes and play an important role in...

Sa1034 Microparticle Dependent Procoagulant Activity and Thrombin Generation is Increased in Patients With Cirrhosis Induced Coagulopathy

Introduction Recent data suggests stable cirrhotics may have a hypercoagulable phenotype. Microparticles (MPs) are submicron plasma particles formed by the exocytic budding of cell membranes and play an important role in haemostasis due to phosphatidylserine (PS) surface expression which provides a phospholipid surface for assembly of coagulation enzymes and/or the expression of tissue factor (TF), the primary initiator of coagulation. To determine whether MPs may contribute to this hypercoagulable phenotype, we assessed microparticle associated functional procoagulant and phenotypic characteristics in cirrhotics. Methods 72 consecutive cirrhotics and 30 healthy volunteers were recruited. Platelet free plasma (PFP) was prepared by two centrifugations and MP-free plasma (MP-FP) by filtration of PFP through a 200 nm microparticle filtration unit. Microparticle (MP) associated procoagulant activity (PCA) was measured using the STA Procoag PPL (phospholipid) assay (Stago Diagnostics) and MP associated thrombin generation (TG) measured using the calibrated automated thrombogram (CAT). For the CAT assay TG was initiated by adding CaCl2 and 1 pM tissue factor, but no phospholipid (PRP reagent), therefore TG was dependent upon phospholipid present in the sample. Flow cytometry (LSRII) was used to determine MP size, number and cellular origin using marker specific antibodies. Results PFP from cirrhotics generated significantly more thrombin than healthy volunteers reflected in the ETP (1374.3 vs 1142.6 nM/min, p=0.04), the peak (101.0 vs 66.5 nM, p=0.001) and a shorter time to peak (13.0 vs 14.2 min, p=0.03). Similarly, MP associated PCA was significantly increased in cirrhotics (65.9±13.2 s), compared to healthy volunteers (74.6±13.9 s, p=0.005). Following filtration of MPs >200 nM in size, there was a large reduction in ETP and peak in both cirrhotics and healthy volunteers, with prolongation of both the time to peak and PPL time. There was significant inverse correlation between the PPL assay and parameters of the TG test [ETP (r=−0.57, p<0.001), Peak (r=−0.43, p<0.001)]. Cirrhotic patients had high levels of Annexin V binding PS positive MPs compared to controls (1412 vs 279 per u/l, p<0.05). Conclusion Microparticle dependent procoagulant activity and thrombin generating capacity is increased in plasma from cirrhotics. High levels of annexin-V positive procoagulant MPs are a likely key and previously undescribed mechanism contributing to the hypercoaguable phenotype observed in cirrhotics. Competing interests None declared.

Size-Dependent Filtration and Trapping of Microparticles in a Microfluidic Chip Using Graduated Gaps and Centrifugal Force

We proposed size-dependent microparticle filtration and trapping using graduated microchannel gaps and centrifugal force using a three-dimensional magnetically driven microtool (3D-MMT) in a microfluidic chip made of polydimethylsiloxane (PDMS). Our paper contributes the following to the field: (1) Particle filtration is robust against pressure fluctuation due to tube vibration between the chip and pump. (2) Clogging by microparticles is avoided by rotating the 3D-MMT in a microchamber. (3) Size-classified microparticles are trapped by flow control along microchannel gaps. Different-sized microparticles flow in spiral microchannels and are filtered based on size between gaps and the substrate by centrifugal force. Microparticles larger than gaps remain in the inner microchannel. Rotating the 3D-MMT using an external magnetic circuit generates swirling flow in the microchamber. Size-classified microparticles are trapped in microchannels by closing the drain port for the targeted particle. Trapped particles are measured by direct observation and treated by reagent. After experiments, trapped particles are extracted by opening drain ports. We demonstrated microparticle filtration and microparticle trapping in the microfluidic chip.