Onset of Nonlinear Electroosmotic Flow under an AC Electric Field.

Nonlinearity of electroosmotic flows (EOFs) is ubiquitous and plays a crucial role in ion transport, specimen mixing, electrochemistry reaction, and electric energy storage and utilization. When and how the transition from a linear regime to a nonlinear one occurs is essential for understanding, prohibiting, or utilizing nonlinear EOF. However, due to the lack of reliable experimental instruments with high spatial and temporal resolutions, the investigation of the onset of nonlinear EOF still remains in theory. Herein, we experimentally studied the velocity fluctuations of EOFs driven by an alternating current (AC) electric field via ultrasensitive fluorescent blinking tricks. The linear and nonlinear AC EOFs are successfully identified from both the time trace and energy spectra of velocity fluctuations. The transitional electric field (EA,C) is determined by both the convection velocity (U) and AC frequency (ff) as EA,C ∼ ff0.48-0.027U. We hope the current investigation could be essential in the development of both theory and applications of nonlinear EOFs.

A Stable NanoPAA-ZnO/ZnCl2 Composite with Variable 3D Structured Morphology and Sustained Superhydrophilicity.

A ZnO/ZnCl2 composite with stable 3D structural morphologies and long lasting superhydrophilicity was synthesized on the top surface of a nano porous anodic alumina (nanoPAA) substrate. The wettability of a nanoPAA-ZnO/ZnCl2 was systematically characterized and the experimental data indicated that the water contact angle (WCA) of 0° could be achieved as well as maintained over 7 days and still remained at 4.36° after 50 days, and its 3D structural morphology had no clearly observable change during this period. The mechanism for the superhydrophilicity of the composites was interpreted in terms of the inherent hydrophilicity of ZnO/ZnCl2 nanofilm, the three-dimensional structures of wrinkled nanoflakes, the nanogaps between neighbor nanoflakes, the difference of structual morphologies (i.e., size, shape, and upright posture of nanoflakes), and the measured True Volume of voids in the nanocomposite. The structural morphologies were mainly determined by the parameters such as the original concentration of precursor ZnCl2 and the pore diameter of nanoPAA substrate. The study proposes a promising superhydrophilic nanomaterial and a cost-effective synthesis method, which will play a practical role in the fields of biomedical molecular sensors and micro/nanofluidic chips.

Artificial Cellulosome Complex from the Self-Assembly of Ni-NTA-Functionalized Polymeric Micelles and Cellulases.

Polymer-protein core-shell nanoparticles have been explored for enzyme immobilization. This work reports on the development of functional polymeric micelles for immobilizing His6 -tagged cellulases with controlled spatial orientation of enzymes, resulting in "artificial cellulosomes" for effective cellulose hydrolysis. Poly(styrene)-b-poly(styrene-alt-maleic anhydride) was prepared through one-pot reversible addition-fragmentation chain-transfer polymerization and modified with nitrilotriacetic acid (NTA) to afford an amphiphilic block copolymer. The self-assembled polymer was mixed with a solution of NiSO4 to form Ni-NTA-functionalized micelles, which could successfully capture His6 -tagged cellulases and form hierarchically structured core-shell nanoparticles with cellulases as the corona. Because the anchored enzymes are site-specifically oriented and in close proximity, synergistic catalysis that results in over twofold activity enhancement has been achieved.

Drug screening assay based on the interaction of intact Keap1 and Nrf2 proteins in cancer cells.

BACKGROUND: The Nrf2-Keap1 interaction is the major regulatory pathway for cytoprotective responses against oxidative and electrophilic stresses. Keap1, a substrate protein of a Cul3-dependent E3 ubiquitin ligase complex, is a negative regulator of Nrf2. The use of chemicals to regulate the interaction between Keap1 and Nrf2 has been proposed as a strategy for the chemoprevention of degenerative diseases and cancers. RESULTS: The interactions between Keap1 and Nrf2 in vitro and in vivo were investigated using fluorescence resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC) strategies in our study. Nrf2 with its N-terminal fused to eGFP and Keap1 with its C-terminal fused to mCherry were expressed and purified in vitro. When purified eGFP-Nrf2 and Keap1-mChrry proteins were mixed together, a strong FRET signal could be detected, indicating an efficient energy transfer from eGFP to mCherry. Moreover, the FRET was detected in vivo using confocal microscopy in colon cancer HCT-116 cells that were co-transfected with eGFP-Nrf2 and Keap1-mCherry. Finally, using an eGFP BiFC approach, the Keap1-Nrf2 interaction was also detected in MCF7 cells by transfecting eGFP N-terminal fused to Nrf2 (eN158-Nrf2) and eGFP C-terminal fused to Keap1 (eC159-Keap1). Using the BiFC and FRET systems, we demonstrated that the prototypical Nrf2-activiting compound tBHQ and the antitumor drug F-dUrd might interfere with the intracellular interaction between Keap1 and Nrf2 whereas the 5-Fu have little role in activating the protective response of Nrf2 pathway in cancer cells. CONCLUSIONS: By analyzing the perturbation of the energy transfer between the donor and acceptor fluorophores and the bimolecular fluorescence complementation of eGFP, we can screen potential inhibitors for the interaction between Keap1 and Nrf2.

Study of Oscillating Electroosmotic Flows with High Temporal and Spatial Resolution.

Near-wall velocity of oscillating electroosmotic flow (OEOF) driven by an AC electric field has been investigated using a laser-induced fluorescence photobleaching anemometer (LIFPA). For the first time, an up to 3 kHz velocity response of OEOF has been successfully measured experimentally, even though the oscillating velocity is as low as 600 nm/s. It is found that the oscillating velocity decays with the forcing frequency ff as ff-0.66. In the investigated range of electric field intensity (EA), below 1 kHz, the linear relation between oscillating velocity and EA is also observed. Because the oscillating velocity at high frequency is very small, the contribution of noise to velocity measurement is significant, and it is discussed in this manuscript. The investigation reveals the instantaneous response of OEOF to the temporal change of electric fields, which exists in almost all AC electrokinetic flows. Furthermore, the experimental observations are important for designing OEOF-based micro/nanofluidics systems.

Studying compaction-decompaction of DNA molecules induced by surfactants.

The mechanism and detailed processes of DNA compaction and decompaction are essential for the life activities, as well as for the researches in the molecular biology, genetics and biomedicine. The compaction of two kinds of DNA molecules caused by Cetyltrimethyl Ammonium Bromide (CTAB) and their decompaction induced with sodium dodecyl sulfate (SDS) or excessive amount of CTAB have been investigated with multiple perspectives such as the UV-VIS spectrophotometry, dynamic light scattering, and zeta potential. The compaction phenomenon of DNA can easily be observed when the CTAB combines with the DNA, not just when the molar ratio QCTAB/QDNA is approximately equal to 1 as the conventional recognition, but also when QCTAB/QDNA <1,DNA can be compacted; Molecular state of DNA is only changed in the conformational structure, but not in the chemical structure. Finally, a model is suggested to help catch on the biophysical mechanism of DNA chain conformational change.

Abnormal Rheological Phenomena in Newtonian Fluids in Electroosmotic Flows in a Nanocapillary.

Abnormal rheological phenomena arising in Tris-borate-ethylenediaminetetraacetic acid solutions (believed to be Newtonian fluids) were observed in direct current electroosmotic flows within a nanocapillary with a diameter of 200 nm under a low electric field of tens of volts per meter. In solutions with different concentrations and pH values, the flow behavior indices of the power-law fluids were calculated on the basis of current-voltage relations. When the electric field intensity was below a critical value of 6.7 V/m, the fluids exhibited dilatant (shear thickening) effects. Fluid viscosity changed with electric field intensity because the near-wall shear rate of an electroosmotic flow changes with electric field intensity via a power-law relation. When the electric field intensity surpasses the critical electric field, the fluid again becomes Newtonian and has constant viscosity. The investigation shows that in nanocapillaries, fluids commonly believed to be Newtonian can become non-Newtonian near walls as a result of strong nanoscale interfacial effects. The results can also improve our understanding of electroosmosis-related transport phenomena in nanofluidics and biomedical science.

Experimental Investigations on Fluorescence Excitation and Depletion of Carbon Dots.

Carbon dots (CDs) can be readily synthesized and utilized as attractive fluorescent probes for a variety of applications. In this study, we have synthesized CDs using a previously published method and characterized their photo-physical properties. The resultant CDs possess prominent photo-stability and short emission wavelength in the violet region. Our study reveals that CDs, with weak photo-bleaching, enable them to be employed to achieve high spatial resolution in stimulated emission depletion (STED) microscopy. The depletion efficiency can reach 60%. More importantly, the shorter excitation wavelength of CDs contributes to further improvement of resolution for STED microscopy. An excellent candidate for fluorophores, these CDs have potential to be used in super-resolution imaging for STED microscopy.

Dynamic behavior of DNA molecules in microchannels: exploring deflective, elliptical, and spin motions induced by Saffman and Magnus forces.

Precise manipulation of individual DNA molecules entering and leaving the channel ports, as well as their smooth passage across the channel, is essential for the detection and screening of DNA molecules using nano-/micro-fluidic technologies. In this paper, by combining single-molecule fluorescence imaging and numerical simulations, the motion states of DNA molecules translocating through a microfluidic channel under the action of the applied electric field are monitored and analyzed in detail. It is found that, under certain conditions of the applied electric field DNA molecules exhibit various motion states, including translation crossing, deflection outflow, reverse outflow, reciprocal movement, and elliptical movement. Simulations indicate that, under the action of Saffman force, DNA molecules can only undergo deflective motion when they experience a velocity gradient in the microchannel flow field; and they can only undergo elliptical motion when their deflective motion is accompanied by a spin motion. In this case, the Magnus force also plays an important role. The detailed study and elucidation of the movement states, dynamic characteristics and mechanisms of DNA molecules such as the deflective and elliptical motions under the actions of Saffman and Magnus forces have helpful implications for the development of related DNA/gene nano-/microfluidic chips, and for the separation, screening and detection of DNA molecules.

An acid catalyzed reversible ring-opening/ring-closure reaction involving a cyano-rhodamine spirolactam.

Cyanamide was introduced into the rhodamine spirolactam framework to produce a colorless and non-fluorescent compound RBCN. It shows a reversible ring-opening/ring-closure process in response to the solution pH, which exhibits an "ON/OFF" switching in its fluorescence. Different from other rhodamine-type dyes, the ring-open form of RBCN is stable in protic solvents under neutral, near neutral and basic conditions, showing a pink color and very strong fluorescence. We also demonstrated the potential of RBCN in live cell imaging.

Cascade and staggered dielectrophoretic cell sorters.

We have developed two new microfluidic cell sorters based on conventional negative dielectrophoresis (DEP) for continuous flow operations. The first is a cascade configuration sorter designed to increase purity of isolated target cell. The second has two staggered side channels in opposite side walls to increase sample throughput without compromising enrichment factor. Particles (carboxylate microspheres) of different sizes were first used to demonstrate the feasibility of the present DEP sorters for cell isolation. Then biological cells, i.e. human prostate cancer cell line LNCaP and human colorectal cancer cell line HCT116 were used to test the performance of the DEP sorters. In the present work, applied voltage was in the range of 0-20 V(p-p) , and frequency was from 0 to 10 MHz. Comparing to a single side channel DEP cell sorter, the isolation purity was improved from 80 to 96% by a single cascade sorter and the sample throughput was increased from 0.2 to 0.65 µL/min by a single staggered side channel sorter. In this article, we report the cell sorter designs, cell separation and enrichment factors.

Rapid AC Electrokinetic Micromixer with Electrically Conductive Sidewalls.

We report a quasi T-channel electrokinetics-based micromixer with electrically conductive sidewalls, where the electric field is in the transverse direction of the flow and parallel to the conductivity gradient at the interface between two fluids to be mixed. Mixing results are first compared with another widely studied micromixer configuration, where electrodes are located at the inlet and outlet of the channel with electric field parallel to bulk flow direction but orthogonal to the conductivity gradient at the interface between the two fluids to be mixed. Faster mixing is achieved in the micromixer with conductive sidewalls. Effects of Re numbers, applied AC voltage and frequency, and conductivity ratio of the two fluids to be mixed on mixing results were investigated. The results reveal that the mixing length becomes shorter with low Re number and mixing with increased voltage and decreased frequency. Higher conductivity ratio leads to stronger mixing result. It was also found that, under low conductivity ratio, compared with the case where electrodes are located at the end of the channel, the conductive sidewalls can generate fast mixing at much lower voltage, higher frequency, and lower conductivity ratio. The study of this micromixer could broaden our understanding of electrokinetic phenomena and provide new tools for sample preparation in applications such as organ-on-a-chip where fast mixing is required.

A novel far-field nanoscopic velocimetry for nanofluidics.

For the first time we have been able to measure the flow velocity profile for nanofluidics with a spatial resolution better than 70 nm. Due to the diffraction resolution barrier, traditional optical methods have so far failed in measuring the velocity profile in a nanocapillary or a closed nanochannel without an opened sidewall. A novel optical point measurement method is presented which applies stimulated emission depletion (STED) microscopy to laser induced fluorescence photobleaching anemometer (LIFPA) techniques to measure flow velocity. Herein we demonstrate this far-field nanoscopic velocimetry method by measuring the velocity profile in a nanocapillary with an inner diameter of 360 nm. The closest measuring point to the wall is about 35 nm. This method opens up a new class of functional measuring techniques for nanofluidics and for nanoscale flows from the wall.

Large-Scale Flow in Micro Electrokinetic Turbulent Mixer.

In the present work, we studied the three-dimensional (3D) mean flow field in a micro electrokinetic (µEK) turbulence based micromixer by micro particle imaging velocimetry (µPIV) with stereoscopic method. A large-scale solenoid-type 3D mean flow field has been observed. The extraordinarily fast mixing process of the µEK turbulent mixer can be primarily attributed to two steps. First, under the strong velocity fluctuations generated by µEK mechanism, the two fluids with different conductivity are highly mixed near the entrance, primarily at the low electric conductivity sides and bias to the bottom wall. Then, the well-mixed fluid in the local region convects to the rest regions of the micromixer by the large-scale solenoid-type 3D mean flow. The mechanism of the large-scale 3D mean flow could be attributed to the unbalanced electroosmotic flows (EOFs) due to the high and low electric conductivity on both the bottom and top surface.

Microfluidic-assisted polymer-protein assembly to fabricate homogeneous functionalnanoparticles.

Functional polymer-protein nanoparticles (NPs) have broad applications in biotechnology and nanotechnology. In principle, controllable and vigorous mixing is required to fabricate homogeneous NPs, which remains a challenge via conventional bulk synthetic methods. In this study, an electrokinetics (EK) based microfluidic reactor with fast mixing is explored to assemble functional proteins with polymers in an ethanol/water co-solvent system. The resultant NPs show significantly improved size distribution by comparison with the ones prepared using conventional bulk method, while the NPs size can be tuned by adjusting the mass ratio of polymer to protein. The functionalities of the assembled proteins are sustained upon the EK based microfluidic mixing, indicating the application potential of our method in the controlled assembly of different functional proteins.

Fluorescent Nanoparticles for Stimulated Emission Depletion Microscopy.

While most of the fluorescent nanoparticles used in stimulated emission depletion (STED) microscopy have a long excitation wavelength, many applications need shorter wavelength fluorophores, which are yet to be developed for STED microscopy applications. Here, three kinds of fluorescent nanoparticles, namely silica nanoparticles (NFv465), fluoro-max blue aqueous fluorescent nanoparticles (FBs) and light yellow nanoparticles (LYs) with short excitation wavelength in violet band have been studied to assess whether they are applicable in STED microscopy. The experimental configuration utilizes a 405 nm continuous wave (CW) laser as excitation beam and a 532 nm CW laser as depletion beam. We compare the photostability, photobleaching and depletion efficiency of three kinds of fluorescent nanoparticles in a series of experiments. Light yellow nanoparticles are proved to be a good candidate as fluorophore in STED microscopy.

Study of the rise time in electroosmotic flow within a microcapillary.

For the first time with a temporal resolution higher than 100 micros, we have successfully measured the risetime of electroosmotic flow (EOF) in a microcapillary using recently developed laser induced fluorescence photobleaching anemometer (LIFPA). Although there are some theoretical estimations in literature about the risetime in microcapillaries, to the best of our knowledge, this has never been experimentally validated in a microcapillary with inner diameter less than 100 microm. The LIFPA with high temporal and spatial resolution described in this paper is capable of measuring risetime distribution radially in a cylindrical microcapillary tube. The experimental results show that the risetime under a pulsed electric field decreases with the reduction of the inner diameter of a microcapillary, and the risetime also increases from the wall to the axis for a given microcapillary. These are qualitatively similar to the theoretical prediction. In addition, the initial pressure driven flow does not appear to affect risetime.

Publisher Correction: A special three-layer step-index fiber for building compact STED systems.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

A special three-layer step-index fiber for building compact STED systems.

Up to now, most of stimulated-emission-depletion (STED) systems were lens-based bulky systems. Exchanging some spatial light paths with optical fiber components will make the systems more flexible and will benefit various fields. A big problem to achieve this goal is that the STED beam generated by the traditional method of bulky systems cannot be maintained in an optical fiber due to its birefringence. In this article, we will introduce a type of special optical fiber. With the special fiber, a dark hollow beam with doughnut-shaped focal spot and a concentric beam with Gaussian-shaped focal spot can be generated at the same time. Parameters of a sample and a compact STED system based on it are demonstrated.

Scaling of velocity and scalar structure functions in ac electrokinetic turbulence.

Electrokinetic (EK) turbulence or electrohydrodynamic (EHD) turbulence has been recently achieved in different fluids under both ac [G. Wang et al., Lab Chip 14, 1452 (2014)10.1039/C3LC51403J; Phys. Rev. E 93, 013106 (2016)10.1103/PhysRevE.93.013106] and dc electric fields [A. Varshney et al., Soft Matter 12, 1759 (2016)10.1039/C5SM02316E]. Here, through dimensional analysis, scaling laws of both velocity and electric conductivity structure functions in the forced cascade region of ac EK turbulence can be predicated (similar to Bolgiano-Obukhov scaling law in turbulent Rayleigh-Bénard convection), in either macroscale or microscale flows. In the forced cascade region, EK force, which relies on the direct cascade of conductivity structures, injects energy directly into a wide spectral region to sustain the flow disturbance. The scaling exponents of the second-order velocity and conductivity structures are 2/5 and 4/5, respectively. In addition to the scaling regions, two characteristic small length scales are derived for both weak and strong electric body forces, respectively. This theoretical investigation can significantly enhance our understanding of EK or EHD turbulence while forced by an ac electric field. It can further broaden our understanding of the forced cascade region of forced turbulence and make the manipulation of the turbulent cascade process more flexible and controllable.