Characterizing Theta-Emitter Generation for Use in Microdroplet Reactions.

Theta emitters are useful for generating microdroplets for rapid-mixing reactions. Theta emitters are glass tips containing an internal septum that separates two channels. When used for mixing, the solutions from each channel are sprayed with mixing occurring during electrospray ionization (ESI) with reaction times on the order of microseconds to milliseconds. Theta emitters of increasing size cause the formation of ESI droplets of increasing size, which require longer times for desolvation and increase droplet lifetimes. Droplets with longer lifetimes provide more time for mixing and allow for increased reaction times prior to desolvation. Because theta emitters are typically produced in-house, there is a need to consistently pull tips with a variety of sizes. Herein, we characterize the effect of pull parameters on the generation of distinct-sized theta emitters using a P-1000 tip puller. Of the examined parameters, the velocity value had the largest impact on the channel diameter. This work also compares the effect of pulling parameters between single-channel and theta capillaries to examine how the internal septum in theta capillaries affects tip pulling. We demonstrate the utility of using theta emitters with different sizes for establishing distinct reaction times. Finally, we offer suggestions on producing theta emitters of various sizes while maintaining high repeatability. Through this work, we provide resources to establish a versatile and inexpensive rapid-mixing system for probing biologically relevant systems and performing rapid derivatizations.

Adjustment of optical absorption in phosphorene through electron-phonon coupling and an electric field.

This study investigates the optical absorption of monolayer phosphorene, focusing on its response to the electron-phonon coupling (EPC) and an electric field. Using a tight-binding Hamiltonian model based on the Barisic-Labbe-Friedel-Su-Schrieffer-Heeger model and the Kubo formula, we calculate the electronic band structure and optical absorption characteristics. The anisotropic dispersion of carriers along armchair and zigzag directions leads to distinct optical responses. Positive and negative EPC effects increase and decrease hopping parameters, respectively, enlarging and reducing/closing the band gap. Moreover, both EPCs cause an admixture of blue and red shift spectrum along the armchair direction, while a red (blue) shift spectrum is observed for positive (negative) EPC along the zigzag direction. Incorporating electric field effects in the EPC increases band gaps for both positive and negative EPC activities, resulting in shifted optical peaks along both directions.

Bacteriophage PVN06 protected catfish Pangasianodon hypophthalmus from Edwardsiella ictaluri infection.

Bacillary necrosis of pangasius (BNP) is a disease caused by Edwardsiella ictaluri bacteria in striped catfish Pangasianodon hypophthalmus that results in high mortality rates. To control this disease, bacteriophages have been considered as alternatives to antibiotics. In this study, we applied the lytic bacteriophage PVN06 in striped catfish fingerlings to prevent E. ictaluri infection. In an experimental trial, the phage was administered to fish by feeding phage-coated feed with doses of 7.17±0.09, 8.17±0.09 and 9.17±0.09 log PFU/g feed per day before bacterial infection. Fish were infected by bacteria once with concentrations ranging from 3.01 to 7.01 log CFU/ml tank water. A day after infection, phage treatment resumed at a rate of once per day until the end of the trial. The results of the trial show that bacterial infection caused typical symptoms of BNP in fish with the cumulative fish death rate of 36.7±2.9 to 75.0±5.0%, depending on the bacterial concentration used for infection. Phage treatment with 9.17±0.09 log PFU/g significantly reduced the mortality rate, while treatments with 8.17±0.09 and 7.17±0.09 log PFU/g did not. This phage dose resulted in a 61.7-fold reduction in the toxicity of the bacterial pathogen and the survival rate of 15-23.3% in fish. Our study has demonstrated that the bacteriophage PVN06 protected striped catfish from BNP.

Complete genome sequence of a novel lytic phage of Xanthomonas oryzae pv. oryzae, the bacterial leaf blight pathogen in rice.

Bacteriophage L522, which infects Xanthomonas oryzae pv. oryzae, was isolated from a paddy leaf sample collected in Long An province, Vietnam. The phage shows myovirus morphology based on transmission electron microscopy. It displays a latent period and burst size of approximately 3 h and 63 new virions per infected cell (PFU/infected cell), respectively. The genome of L522 is 44,497 bp in length, with 52% GC content. Of the 63 genes identified, functions were predicted for 26. No virulence or antibiotic-resistance genes were detected. The results of a BLASTn search showed similarity to a previously reported Xanthomonas phage, with 85% average nucleotide sequence identity and 87.15% query coverage. Thus, this L522 is a representative of a new species in the genus Xipdecavirus.

Bacterial chemotaxis in static gradients quantified in a biopolymer membrane-integrated microfluidic platform.

Chemotaxis is a fundamental bacterial response mechanism to changes in chemical gradients of specific molecules known as chemoattractant or chemorepellent. The advancement of biological platforms for bacterial chemotaxis research is of significant interest for a wide range of biological and environmental studies. Many microfluidic devices have been developed for its study, but challenges still remain that can obscure analysis. For example, cell migration can be compromised by flow-induced shear stress, and bacterial motility can be impaired by nonspecific cell adhesion to microchannels. Also, devices can be complicated, expensive, and hard to assemble. We address these issues with a three-channel microfluidic platform integrated with natural biopolymer membranes that are assembled in situ. This provides several unique attributes. First, a static, steady and robust chemoattractant gradient was generated and maintained. Second, because the assembly incorporates assembly pillars, the assembled membrane arrays connecting nearby pillars can be created longer than the viewing window, enabling a wide 2D area for study. Third, the in situ assembled biopolymer membranes minimize pressure and/or chemiosmotic gradients that could induce flow and obscure chemotaxis study. Finally, nonspecific cell adhesion is avoided by priming the polydimethylsiloxane (PDMS) microchannel surfaces with Pluronic F-127. We demonstrated chemotactic migration of Escherichia coli as well as Pseudomonas aeruginosa under well-controlled easy-to-assemble glucose gradients. We characterized motility using the chemotaxis partition coefficient (CPC) and chemotaxis migration coefficient (CMC) and found our results consistent with other reports. Further, random walk trajectories of individual cells in simple bright field images were conveniently tracked and presented in rose plots. Velocities were calculated, again in agreement with previous literature. We believe the biopolymer membrane-integrated platform represents a facile and convenient system for robust quantitative assessment of cellular motility in response to various chemical cues.

Ferritin Conjugates With Multiple Clickable Amino Acids Encoded by C-Terminal Engineered Pyrrolysyl-tRNA Synthetase.

This study reports the application of expanding genetic codes in developing protein cage-based delivery systems. The evolved Methanosarcina mazei pyrrolysyl-tRNA synthetase (PylRS)•tRNAPyl pairs derived from directed evolution are examined to probe their recognition for para-substituted phenylalanine analogs. The evolved MmPylRS, AzFRS, harboring a wide range of substrates, is further engineered at the C-terminal region into another variant, AzFRS-MS. AzFRS-MS shows suppression of the elevated sfGFP protein amount up to 10 TAG stop codons when charging p-azido-l-phenylalanine (AzF, 4), which allows the occurrence of click chemistry. Since protein nanocages used as drug delivery systems that encompass multiple drugs through a site-specific loading approach remain largely unexplored, as a proof of concept, the application of AzFRS-MS for the site-specific incorporation of AzF on human heavy chain ferritin (Ftn) is developed. The Ftn-4 conjugate is shown to be able to load multiple fluorescence dyes or a therapeutic agent, doxorubicin (Dox), through the strain-promoted azide-alkyne cycloaddition (SPAAC) click reaction. Aiming to selectively target Her2+ breast cancer cells, Ftn-4-DOX conjugates fused with a HER2 receptor recognition peptide, anti-Her2/neu peptide (AHNP), is developed and demonstrated to be able to deliver Dox into the cell and to prolong the drug release. This work presents another application of evolved MmPylRS systems, whose potential in developing a variety of protein conjugates is noteworthy.

Diverse Bacteriophages Infecting the Bacterial Striped Catfish Pathogen Edwardsiella ictaluri.

Bacteriophages infecting Edwardsiella ictaluri have been less investigated, although the host bacterium is one of the most important fish pathogens causing enteric septicemia of catfish (ESC). We present here two distinctly novel bacteriophages vB_EiM_PVN06 and vB_EiA_PVN09 infecting Edwardsiella ictaluri E1, with their geographical origins from the Mekong Delta, Vietnam. Bacteriophage vB_EiM_PVN06 native to a mud sample reveals complete differences of biological properties with the phage vB_EiA_PVN09 originated from a viscus of a healthy catfish (Pangasianodon hypophthalmus) cultured in the same area. Morphological analyses combined with genomic data indicate that phage vB_EiM_PVN06 is classified to Myoviridae family and shares high similarity with E. ictaluri phage PEi21 genome, while vB_EiA_PVN09 is a member of Teseptimavirus genus, Autographiviridae family, and mostly closes to phage vB_EcoP_IME390. The vB_EiA_PVN09 is a T7-like bacteriophage, which has been firstly found infecting to E. ictaluri, and host range analysis also evidences for the cross-infection of this phage to Escherichia coli K12 and Escherichia coli DH5α. Together, our research highlights the diversity of bacteriophages infecting the pathogen E. ictaluri and suggests further explorations of lytic phages in environmental niches, to be exploited in feasible strategies of phage therapy in ESC disease control.

CometChip enables parallel analysis of multiple DNA repair activities.

DNA damage can be cytotoxic and mutagenic, and it is directly linked to aging, cancer, and other diseases. To counteract the deleterious effects of DNA damage, cells have evolved highly conserved DNA repair pathways. Many commonly used DNA repair assays are relatively low throughput and are limited to analysis of one protein or one pathway. Here, we have explored the capacity of the CometChip platform for parallel analysis of multiple DNA repair activities. Taking advantage of the versatility of the traditional comet assay and leveraging micropatterning techniques, the CometChip platform offers increased throughput and sensitivity compared to the traditional comet assay. By exposing cells to DNA damaging agents that create substrates of Base Excision Repair, Nucleotide Excision Repair, and Non-Homologous End Joining, we show that the CometChip is an effective method for assessing repair deficiencies in all three pathways. With these applications of the CometChip platform, we expand the utility of the comet assay for precise, high-throughput, parallel analysis of multiple DNA repair activities.

CometChip analysis of human primary lymphocytes enables quantification of inter-individual differences in the kinetics of repair of oxidative DNA damage.

Although DNA repair is known to impact susceptibility to cancer and other diseases, relatively few population studies have been performed to evaluate DNA repair kinetics in people due to the difficulty of assessing DNA repair in a high-throughput manner. Here we use the CometChip, a high-throughput comet assay, to explore inter-individual variation in repair of oxidative damage to DNA, a known risk factor for aging, cancer and other diseases. DNA repair capacity after H2O2-induced DNA oxidation damage was quantified in peripheral blood mononuclear cells (PBMCs). For 10 individuals, blood was drawn at several times over the course of 4-6 weeks. In addition, blood was drawn once from each of 56 individuals. DNA damage levels were quantified prior to exposure to H2O2 and at 0, 15, 30, 60, and 120-min post exposure. We found that there is significant variability in DNA repair efficiency among individuals. When subdivided into quartiles by DNA repair efficiency, we found that the average t1/2 is 81 min for the slowest group and 24 min for the fastest group. This work shows that the CometChip can be used to uncover significant differences in repair kinetics among people, pointing to its utility in future epidemiological and clinical studies.

Three-dimensional microscale hanging drop arrays with geometric control for drug screening and live tissue imaging.

Existing three-dimensional (3D) culture techniques are limited by trade-offs between throughput, capacity for high-resolution imaging in living state, and geometric control. Here, we introduce a modular microscale hanging drop culture where simple design elements allow high replicates for drug screening, direct on-chip real-time or high-resolution confocal microscopy, and geometric control in 3D. Thousands of spheroids can be formed on our microchip in a single step and without any selective pressure from specific matrices. Microchip cultures from human LN229 glioblastoma and patient-derived mouse xenograft cells retained genomic alterations of originating tumors based on mate pair sequencing. We measured response to drugs over time with real-time microscopy on-chip. Last, by engineering droplets to form predetermined geometric shapes, we were able to manipulate the geometry of cultured cell masses. These outcomes can enable broad applications in advancing personalized medicine for cancer and drug discovery, tissue engineering, and stem cell research.

SpheroidChip: Patterned Agarose Microwell Compartments Harboring HepG2 Spheroids are Compatible with Genotoxicity Testing.

Three-dimensional tissue culture models are emerging as effective alternatives to animal testing. They are especially beneficial for liver toxicity studies, enabling hepatocytes to display improved levels of liver-specific functions. One common model is hepatocyte spheroids, which are spontaneously formed cell aggregates. Techniques for spheroid formation include the use of ultralow attachment plates and the hanging drop method, both of which are technically challenging and relatively low throughput. Here, we describe a simple-to-use platform that improves spheroid production and is compatible with genotoxicity testing by the comet assay. To achieve this, we created a chip containing a microwell array where dozens of spheroids are contained within a single well of a 96-well plate. The microwells are made from agarose, a nontoxic material suitable for cell growth and spheroid formation. HepG2 cells loaded into customizable microwells formed spheroids through agarose-assisted aggregation within one to two days. In addition, the agarose matrix allows the same platform to be used in DNA damage analysis. Specifically, the comet assay enables quantification of DNA breaks based on the increased migration of damaged DNA through agarose during electrophoresis. Here, we developed a modified comet assay and show that intact HepG2 spheroids cultured in microwells can be electrophoresed to reveal the extent of DNA damage following exposure to inflammatory chemicals (H2O2 and SIN-1). With this SpheroidChip analysis method, we detected a dose-dependent increase in DNA damage and observed rapid repair of H2O2-induced DNA damage. In summary, we utilized an agarose microarray to condense what had required an entire 96-well plate into a single well, enabling analysis techniques that were cumbersome or impossible under conditions of a single spheroid per well of a 96-well plate.

Effect of increasing inclusion rates of tofu by-product in diets of growing pigs on nitrogen balance and ammonia emission from manure.

To reduce competition with human-edible feed resources, it is of interest to incorporate by-products from the food industry in animal feeds. The current research investigated the effect of including increasing amounts of tofu by-product (TF) in practical pig diets on animal performance, nitrogen balance and ammonia emissions from manure. Two experiments were conducted including a control diet without TF, containing 160 g/kg dietary non-starch polysaccharides (NSPs) and three diets including 122, 246 and 360 g TF/kg DM (TF122, TF246 and TF360, respectively) to reach 220, 280 and 360 g/kg NSP. All diets had the same level of CP and protein digestible in the small intestine which particularly was realized by replacing rice bran with TF. Animal performance was assessed in a first experiment with 40 growing barrows with initial BW of 26.6 ± 1.80 kg (M ± SD) being allocated to the 4 treatments, during 2 growth phases (i.e. until 50 kg BW and from 50 to 80 kg BW). In the growth phase until 50 kg, feed intake and average daily gain (ADG) were linearly reduced by dietary TF inclusion, while this negative impact disappeared during the second growth phase (50 to 80 kg BW). Tofu by-product inclusion even positively affected the feed conversion ratio during this second growth phase (3.4 to 2.7 kg feed/kg ADG for 0 to 360 g/kg dietary TF). Over the entire growth period, performance and feed intake were negatively affected at the highest dietary TF level. Experiment 2 was conducted to assess digestibility, nitrogen balance and ammonia emission from manure. For this purpose, 16 pigs with BW of 62.8 ± 3.6 kg (M ± SD) were assigned to either 1 of the 4 treatments. There was no difference in total tract apparent digestibility of dietary organic matter or CP, while NDF digestibility increased with increasing TF level, suggesting increasing importance of the hindgut fermentation when digesting diets with increasing TF levels. Nevertheless, this was not reflected in increasing levels of faecal volatile fatty acids or purines, nor in reduced manure pH. As a result, ammonia emission from slurry was not reduced through dietary TF inclusion, despite the linear decrease in urinary nitrogen. In conclusion, TF can be included in pigs' diets up to an inclusion rate of 25% without risk of impaired animal performance; however, this dietary strategy fails to mitigate ammonia emission from slurry.

Sensitive CometChip assay for screening potentially carcinogenic DNA adducts by trapping DNA repair intermediates.

Genotoxicity testing is critical for predicting adverse effects of pharmaceutical, industrial, and environmental chemicals. The alkaline comet assay is an established method for detecting DNA strand breaks, however, the assay does not detect potentially carcinogenic bulky adducts that can arise when metabolic enzymes convert pro-carcinogens into a highly DNA reactive products. To overcome this, we use DNA synthesis inhibitors (hydroxyurea and 1-ß-d-arabinofuranosyl cytosine) to trap single strand breaks that are formed during nucleotide excision repair, which primarily removes bulky lesions. In this way, comet-undetectable bulky lesions are converted into comet-detectable single strand breaks. Moreover, we use HepaRG™ cells to recapitulate in vivo metabolic capacity, and leverage the CometChip platform (a higher throughput more sensitive comet assay) to create the 'HepaCometChip', enabling the detection of bulky genotoxic lesions that are missed by current genotoxicity screens. The HepaCometChip thus provides a broadly effective approach for detection of bulky DNA adducts.

ß12-Borophene becomes a semiconductor and semimetal via a perpendicular electric field and dilute charged impurity.

In this paper, the possible electronic phase transitions of ß12-borophene crystal are examined using a five-band tight-binding calculation. For different tight-binding models, the Green's function technique is employed for the electronic density of states (DOS). We focus on the modulation of the DOS around the Fermi level with a perpendicular electric field and the dilute charged impurity. The steps to incorporate the effects of external electric field and charged impurity are also detailed with the local Hamiltonian model and the Born approximation, respectively. Our calculations show that the inversion symmetric model is the proper model to discuss the metallic phase of the system, entailing different results compared to the homogeneous model. We find that the electric field opens a tunable band gap and a metal-to-p-doped semiconductor phase transition emerges at the strong perpendicular electric field. The influence of impurity scattering potential on the electronic phase of ß12-borophene is much larger than the impurity concentration, in which a metal-to-n-doped semiconductor (metal-to-semimetal) transition takes place at high scattering potentials for the homogeneous (inversion symmetric) model, whereas there is no transition when the impurity concentration is changed. Thereby, producing semimetallic/semiconducting properties by applying an appropriate external electric field and dilute charged impurities paves the way for the realization of ß12-borophene-based nano-optoelectronic devices.

Does software optimization influence the radiologists' perception in low dose paediatric pelvic examinations?

PURPOSE: To investigate whether software optimisation can improve an observers' perception of image quality in low dose paediatric pelvic examinations. METHODS: Twenty-five consecutive patients (3-7 years old) were referred for a pelvic digital radiography (DR) examination. They were prospectively enrolled in the study over a 3-month period. Images were taken at 80 kV and 2-4 mAs depending on pelvic thickness (9-15 cm). A small focal spot, 130 cm SID: 10 cm air gap and 1 mm Al and 0.2 mm Cu additional filtration were also utilised. Images were acquired on a Canon DR detector and optimised using five different combinations of the multi-frequency processing software (Canon DR system version NE, Version 7.1 with SPECTRA) to comply with the ALARA principle. Image quality was blindly evaluated using the subjective Visual Grading Analysis (VGA) by five experienced musculoskeletal radiologists, including the evaluation of six anatomical image quality criteria (scored from 1 to 5). RESULTS: Consistently, the VGA results indicated that by using software optimised parameters, image quality was suitable for diagnosis in 48-71% of all images. Based on a VGC analysis all software optimised images did have significant better image quality then the one with just the clinical settings. Noise reduction was the software setting which influenced the image quality the most, area under the curve (AUC) of 0.8172 95%CI 0.7953-0.8375. CONCLUSION: Software optimisation improve the radiologists' perception of image quality and should thus be thoroughly considered within clinical practise. Noise reduction is the software parameter which has the greatest influence.

Selection of Phages to Control Aeromonas hydrophila - An Infectious Agent in Striped Catfish.

Striped catfish (Pangasianodon hypophthalmus) farming in the Mekong Delta Vietnam (MKDVN) importantly contributes to national aquaculture export. Currently, however, diseases occur more frequently across the entire MKDVN region. One of the most common types is hemorrhagic septicemia caused by Aeromonas hydrophila. In this study, isolation and selection of the phages for control in vitro Aeromonas hydrophila were conducted. 24 phages were isolated from 100 striped catfish pond water samples. Next, lytic activity of these phages was clarified. Four phages with short latent period (about 25 to 40 min) and/or high burst size (about 67 to 94 PFU/ cell) were selected to evaluate their infection activity to different phage-resistant A. hydrophila strains. Two phages termed as TG25P and CT45P were subjected to the phage cocktail to inactivate A. hydrophila. Re-growth of the host bacteria appeared about eight hours after treatment. Usage of the phage cocktail that attach different host bacterial receptors is not always much effective than usage of single phage. This is the first report about phage therapy to control A. hydrophila isolated from striped catfish. Some challenges in the phage cocktail were shown to achieve strategies in prospective studies in the context of high antibiotic resistance of A. hydrophila.

Microcolony Size Distribution Assay Enables High-Throughput Cell Survival Quantitation.

Cell survival is a critical and ubiquitous endpoint in biology. The broadly accepted colony formation assay (CFA) directly measures a cell's ability to divide; however, it takes weeks to perform and is incompatible with high-throughput screening (HTS) technologies. Here, we describe the MicroColonyChip, which exploits microwell array technology to create an array of colonies. Unlike the CFA, where visible colonies are counted by eye, using fluorescence microscopy, microcolonies can be analyzed in days rather than weeks. Using automated analysis of microcolony size distributions, the MicroColonyChip achieves comparable sensitivity to the CFA (and greater sensitivity than the 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide [XTT] assay). Compared to CellTiter-Glo, the MicroColonyChip is as sensitive and also robust to artifacts caused by differences in initial cell seeding density. We demonstrate efficacy via studies of radiosensitivity and chemosensitivity and show that the approach is amenable to multiplexing. We conclude that the MicroColonyChip is a rapid and automated alternative for cell survival quantitation.

Magneto-EELS of armchair boronitrene nanoribbons.

The evolution of the electron energy loss spectrum (EELS) of ultranarrow armchair boron nitride nanoribbons (aBNNRs) during low and high photon energy transfers has been studied theoretically when a magnetic field and temperature gradient are applied. In order to achieve this goal, the widely used linear response theory within the Green's function theory was employed. Here, using the EELS we show that σ ↦ σ* or π ↦ π* and σ ↦ π* or π ↦ σ* excitations corresponding to the intraband and interband transitions, respectively, can be tuned by ribbon width, magnetic field, wave vector transfer, and temperature. A comparison with experimental studies reveals that for realistic ribbon widths, i.e. 10-100 nm, both excitations are weak. However, we observe that only transitions between the same states, i.e. σ ↦ σ* or π ↦ π* can be controlled with a magnetic field due to the localized highest occupied and lowest unoccupied states at low-energy regions and different states are not influenced when the magnetic field is applied. Interestingly, the detailed shape of the magneto-EELS of the 7-aBNNR indicates a direct-to-indirect band gap transition when the wave vector transfer is perpendicular to the 7-aBNNR plane. Finally, we discover that there is an anomalous behavior for the temperature dependence of the magneto-EELS in general. The present work brings forward the understanding of the magneto-EELS of ultranarrow aBNNRs under different environmental conditions for logic applications in nanoplasmonics.

Interplay of orbital hopping and perpendicular magnetic field in anisotropic phase transitions for Bernal bilayer graphene and hexagonal boron-nitride.

We theoretically address the perpendicular magnetic field effects on the electronic phase of Bernal bilayer graphene and hexagonal boron-nitride (h-BN) taking into account the total and orbital-projected electronic bands using the tight-binding parameters in the Harrison model, followed by the Green's function method. First, we confirm that our model is computationally efficient and accurate for calculating the magneto-orbital electronic phase transition by reproducing the semimetallic and insulating treatments of pristine Bernal bilayer graphene and h-BN, respectively. In our model, the magnetic field couples only to the electron spin degrees of freedom (with the same contributions for spin-up and spin-down) due to the low dimension of the systems. Here, the main features of the phase transitions are characterized by the electronic density of states (DOS). We found that sp2-hybridization is destroyed when the systems are immersed in the magnetic field, leading to a phase transition to metal for both systems at strong magnetic fields. While there is no phase transition for bilayer graphene at weak magnetic fields, for the case of bilayer h-BN, an insulator to semiconductor phase transition can be viewed, making h-BN more applicable in industry. In bilayer graphene, the anisotropic phase transition appears as insulator-semiconductor, insulator-metal, and semimetal-metal for s-, {px + py}-, and pz-orbitals, respectively, whereas in the case of bilayer h-BN, one observes the same transitions for {s,pz}-orbitals but insulator-semiconductor for {px + py} orbitals. Generically, our findings highlight that the applied magnetic field manipulates the band structure of bilayer graphene and h-BN, and gives ideas to experimentalists for tuning the electro-optical properties of these materials.