Endocytosis of a single mesoporous silica nanoparticle into a human lung cancer cell observed by differential interference contrast microscopy

The unique structural features of mesoporous silica nanoparticles (MSN) have made them very useful in biological applications, such as gene therapy and drug delivery. Flow cytometry, confocal microscopy, and electron microscopy have been used for observing the endocytosis of MSN. However, flow cytometry cannot directly observe the process of endocytosis. Confocal microscopy requires fluorescence labeling of the cells. Electron microscopy can only utilize fixed cells. In the present work, we demonstrate for the first time that differential interference contrast (DIC) microscopy can be used to observe the entire endocytosis process of MSN into living human lung cancer cells (A549) without fluorescence staining. There are three physical observables that characterize the locations of MSN and the stages of the endocytosis process: motion, shape, and vertical position. When it was outside the cell, the MSN underwent significant Brownian motion in the cell growth medium. When it was trapped on the cell membrane, the motion of the MSN was greatly limited. After the MSN had entered the cell, it resumed motion at a much slower speed because the cytoplasm is more viscous than the cell growth medium and the cellular cytoskeleton networks act as obstacles. Moreover, there were shape changes around the MSN due to the formation of a vesicle after the MSN had been trapped on the cell membrane and prior to entry into the cell. Finally, by coupling a motorized vertical stage to the DIC microscope, we recorded the location of the MSN in three dimensions. Such accurate 3D particle tracking ability in living cells is essential for studies of selectively targeted drug delivery based on endocytosis.     

Differential interference contrast microscopy for real-time dynamics and manipulation of single cells in microchannels

Nomarski differential interference contrast (DIC) microscopy was used for real-time dynamics of intact single cells in various microchannels for adaptation to microfluidic chip application. The cheek cell was chosen as a model, single cell and the dynamics was measured at the microchannels. The image resolution of single cell was shaper and more distinct in DIC than in conventional microscopy. The individual single living cells were also manipulated by both hydrodynamic and electrokinetic flow-driving forces at the microchannels. The DIC contrast was enhanced according to the order of round-, square-, and rectangle-type microchannels. The velocity of the single living cell was consistently increased with increasing electric field strength and pH. However, the velocity of cell was decreased with increasing run buffer concentration. The driving direction of the individual single cell was simply controlled by changing the polarity of the applied voltage and the electric field strength. The cells were consistently manipulated in the microchannel under the co-application of the low electric field of 2.44 V/cm, instead of the solo application of the hydrodynamic force.     

Effect of instant controlled pressure drop treatments on the oligosaccharides extractability and microstructure of Tephrosia purpurea seeds

The study of the oligosaccharides extracted from Tephrosia purpurea seeds was undertaken using the instant controlled pressure drop (DIC) as a pre-treatment prior to conventional solvent extraction. This DIC procedure provided structural modification in terms of expansion, higher porosity and improvement of specific surface area; diffusion of solvent inside such seeds and availability of oligosaccharides increase notably. In this paper, we investigated and quantified the impact of the different DIC operative parameters on the yields of ciceritol and stachyose extracted from T. purpurea seeds. The treatment could be optimized with a steam pressure (P) (P=0.2 MPa), initial water content (W) (W=30% dry basis (DB)) and thermal treatment time (t) (t=30s). By applying DIC treatment in these conditions, the classic process of extraction was intensified in both aspects of yields (145% of ciceritol and 185% of stachyose), and kinetics (1h of extraction time instead of 4h for conventional process). The scanning electron microscopy micrographs provided evident modifications of structure of seeds due to the DIC treatment.     

Resolving rotational motions of nano-objects in engineered environments and live cells with gold nanorods and differential interference contrast microscopy

Gold nanorods are excellent orientation probes due to their anisotropic optical properties. Their dynamic rotational motion in the 3D space can be disclosed with Nomarski-type differential interference contrast (DIC) microscopy. We demonstrate that by using the combination of gold nanorod probes and DIC microscopy, we are able to resolve rotational motions of nano-cargos transported by motor proteins at video rate not only on engineered surfaces but also on cytoskeleton tracks in live cells.     

Study of frictional properties of a phospholipid bilayer in a liquid environment with lateral force microscopy as a function of NaCl concentration

Friction properties of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC)-supported planar bilayers deposited on mica were tested in a liquid environment by lateral force microscopy. The presence of these bilayers was detected by imaging and force measurements with atomic force microscopy. To test how the presence of NaCl affects the frictional properties of the phospholipid bilayers, four DMPC bilayers were prepared on mica in saline media ranging from 0 to 0.1 M NaCl. Changes in the lateral vs vertical force curves were recorded as a function of NaCl concentration and related to structural changes induced in the DMPC bilayer by electrolyte ions. Three friction regimes were observed as the vertical force exerted by the tip on the bilayer increased. To relate the friction response to the structure of the DMPC bilayer, topographic images were recorded at the same time as friction data. Ions in solution screened charges present in DMPC polar heads, leading to more compact bilayers. As a consequence, the vertical force at which the bilayer broke during friction experiments increased with NaCl concentration. In addition, the topographic images showed that low-NaCl-concentration bilayers recover more easily due to the low cohesion between phospholipid molecules.     

Electrosynthesis and analytical characterization of PMMA coatings on titanium substrates as barriers against ion release

The performance of polyacrylic coatings as barrier films against corrosion of titanium-based orthopaedic implants was investigated. In particular, poly(methyl methacrylate) (PMMA) was electrosynthesized on titanium substrates by electro-reductive processes from aqueous monomer solutions. The obtained PMMA coatings were characterized by Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). The effect of an annealing treatment on the morphology of coatings with respect to uniformity and porosity of films was assessed by scanning electron microscopy (SEM). An inductively coupled plasma-mass spectrometry (ICP-MS) technique was used for ion concentration measurements in ion release tests performed on TiAlV sheets modified with PMMA coatings (annealed and unannealed). Results indicated that the annealing process produces coatings with considerable anticorrosion performances.Electronic Supplementary Material Supplementary material is available for this article at     

碱金属团簇碰撞动力学研究

With in the framework of distance dependent tight-binding molecular dynamics (DDTBMD), the collision dynamics of sodium cluster Na n has been studied systematically. Some phenomena have been observed at different impact parameters b , such as fusion reaction, deep inelastic collision (DIC) and quasi-elasticcollision, which are similar to the nuclear heavy-ion collisions (HIC). For the system of Na6 (3D) + Na8, the reaction mechanism at b=9a0 is DIC, but when b is equal to 13a0, it corresponds to quasi-elastic collision. Further more the rotation processes during the collisions, are related to the collision energy and parameter. The larger collision energy is, the earlier relative rotation will occur, and the relaxation time becomes shorter and the relative rotation energy is much smaller. There exists maximal relative rotation energy which corresponding to b = 7a0. When b is smaller than 7a0, the rotation energy increases with b increasing, otherwise the energyd ecreases. And the maximal relative rotation energy is corresponding to DIC process. The maximal rotation energy can reach on etenth of total energy, which is much less than that in HIC.     

Suwannee河腐植酸光化学生成溶解无机碳的研究

溶解有机质矿化为溶解无机碳是全球碳循环中的一个重要过程.利用模拟太阳光对Suwannee河腐植酸的光氧化过程模拟,研究了溶解氧浓度、模拟太阳光波长范围和铁浓度对溶解无机碳产量的影响.研究表明,基于前24 h空气饱和的无机碳矿化速率4.40μmol/(L.h),氧气饱和条件下照射增加1.56倍,氮气饱和条件下照射下降36%.UV-B,UV-A和可见光部分分别占溶解无机碳产量的31.8%、32.6%和25.6%.当加入20μmol/L铁络合剂DFOM时(初始样品Suwannee河腐植酸铁含量为5.46μmol/L),溶解无机碳产量下降55.6%.     

Three-dimensional imaging of sulfides in silicate rocks at submicron resolution with multiphoton microscopy

We report the first application of multiphoton microscopy (MPM) to generate three-dimensional (3D) images of natural minerals (micron-sized sulfides) in thick (～120 μm) rock sections. First, reflection mode (RM) using confocal laser scanning microscopy (CLSM), combined with differential interference contrast (DIC), was tested on polished sections. Second, two-photon fluorescence (TPF) and second harmonic signal (SHG) images were generated using a femtosecond-laser on the same rock section without impregnation by a fluorescent dye. CSLM results show that the silicate matrix is revealed with DIC and RM, while sulfides can be imaged in 3D at low resolution by RM. Sulfides yield strong autofluorescence from 392 to 715 nm with TPF, while SHG is only produced by the embedding medium. Simultaneous recording of TPF and SHG images enables efficient discrimination between different components of silicate rocks. Image stacks obtained with MPM enable complete reconstruction of the 3D structure of a rock slice and of sulfide morphology at submicron resolution, which has not been previously reported for 3D imaging of minerals. Our work suggests that MPM is a highly efficient tool for 3D studies of microstructures and morphologies of minerals in silicate rocks, which may find other applications in geosciences.     

Determination of the number of cells in preimplantation embryos by using noninvasive optical quadrature microscopy in conjunction with differential interference contrast microscopy.

The number of cells in a preimplantation embryo is directly correlated to the health and viability of the embryo. There are currently no methods to count the number of cells in late-stage preimplantation embryos noninvasively. We assessed the ability of optical quadrature microscopy (OQM) to count the number of cells in mouse preimplantation embryos noninvasively. First, to test for possible light toxicity, we exposed two-cell mouse embryos to OQM and differential interference contrast (DIC) microscopy and assessed their ability to develop to the blastocyst stage. We found no inhibition of development from either mode of microscopy for up to 2 h of light exposure. We also imaged eight-cell to morula stage mouse preimplantation embryos by OQM nd developed two methods for counting the number of cells. The contour signature method (CSM) used OQM images alone and the phase subtraction method (PSM) used both OQM and DIC images. We compared both methods to standard cell counting techniques and found that the PSM was superior to all other noninvasive cell counting methods. Our work on mouse embryos should be applicable to human embryos. The ability to correctly count the number of cells in human preimplantation embryos could lead to the transfer of fewer embryos in in vitro fertilization (IVF) clinics and consequently a lower rate of high-risk multiple-infant births.     

Extraction of essential oils from Algerian myrtle leaves using instant controlled pressure drop technology

In the present work, the new extraction process of Détente Instantanée Contrôlée DIC (French, for instant controlled pressure drop) was studied, developed, quantitatively and qualitatively compared to the conventional hydrodistillation method for the extraction of essential oils from Algerian myrtle leaves. DIC was used as a thermomechanical treatment, DIC subjecting the product to a high-pressure saturated steam. The DIC cycle ends with an abrupt pressure drop towards vacuum, and this instantly leads to an autovaporization of myrtle volatile compounds. An immediate condensation in the vacuum tank produced a micro-emulsion of water and essential oils. Thus, an ultra-rapid cooling of residual leaves occurred, precluding any thermal degradation. An experimental protocol was designed with 3 independent variables: saturated steam pressure between 0.1 and 0.6 MPa, resulting in a temperature between 100 and 160 °C, a total thermal processing time between 19 and 221 s, and between 2 and 6 DIC cycles. The essential oils yield was defined as the main dependent variable. This direct extraction gave high yields and high quality essential oil, as revealed by composition and antioxidant activity (results not shown). After this treatment, the myrtle leaves were recovered and hydrodistilled in order to quantify the essential oil content in residual DIC-treated samples. Scanning electron microscope (SEM) showed some modification of the structure with a slight destruction of cell walls after DIC treatment.     

Stable Diindeno‐Fused Corannulene Regioisomers with Open‐Shell Singlet Ground States and Large Diradical Characters

The synthesis of open‐shell polycyclic hydrocarbons with large diradical characters is challenging because of their high reactivities. Herein, two diindeno‐fused corannulene regioisomers DIC‐1 and DIC‐2 , curved fragments of fullerene C₁₀₄, were synthesized that exhibit open‐shell singlet ground states. The incorporation of the curved and non‐alternant corannulene moiety within diradical systems leads to significant diradical characters as high as 0.98 for DIC‐1 and 0.89 for DIC‐2 . Such high diradical characters can presumably be ascribed to the re‐aromatization of the corannulene π system. Although the DIC compounds have large diradical characters, they display excellent stability under ambient conditions. The half‐lives are 37 days for DIC‐1 and 6.6 days for DIC‐2 in solution. This work offers a new design strategy towards diradicaloids with large diradical characters yet maintain high stability.     

The thermochemical behaviour of zeolite inclusion complexes

The thermochemical behaviour of nitrate inclusion complexes of zeolites of type A was investigated and DTA curves recorded. An attempt was made to identify thermal effects. Supplementary information was obtained from the IR spectra of the solid phase before and after heating as well as from the process of denitration of inclusion complexes.     

气相色谱-稳定同位素质谱法测定溶解无机碳碳同位素

选用NaHCO3配制了浓度分别为0.24、1.19、2.38和4.76 mmol/L的溶解无机碳(D IC)溶液,经过1h、4h、8h和24h不同平衡时间,建立了一种分析D IC碳同位素的方法。不同浓度的D IC样品与其母质NaHCO3的1δ3C值之间的差值仅为(0.2~0.5)‰。通过对照组的实验结果和对空气CO2的碳同位素测试与研究,证明此方法可有效避免实验过程中大气等物质对样品的污染,确保实验结果的准确性。     

Fracture mechanics of laser sintered cracked polyamide for a new method to induce cracks by additive manufacturing

This paper presents an experimental investigation on specimens manufactured by Selective Laser Sintering (SLS), with the purposes of giving designers advice when designing 3D printed parts, and laying the basis for a step forward in the field of fracture mechanics of 3D complex parts.The aim is to investigate the effect of building direction in Polyamide (PA) 3D printed samples and to assess whether a crack can be initiated directly from the sintering process for fracture mechanics study purposes.Six different configurations of Mode I Compact Tension (CT) specimens were manufactured and tested; the experiments were monitored by Digital Image Correlation (DIC) and fractured surfaces were analyzed using microscopy.Results showed that samples with better mechanical performance are those in which all the layers contain a portion of the crack. On the other hand, those with layers parallel to the crack plan offer a preferential pathway for the crack to propagate. DIC and fractography investigations showed that, under certain conditions, small-radius geometries, or too-close surfaces may bond together depending on printer resolution. Experiments also showed that SLS is capable of printing specimens with internal cracks that can be used to study fracture mechanics of complex parts or parts with internal cracks.     

A PDMS-based biochip with integrated sub-micrometre position control for TIRF microscopy of the apical cell membrane

A poly(dimethylsiloxane) (PDMS)-based biochip with an integrated pressure controlled positioning system with sub-micrometre precision was realized. The biochip was easy and cheap to manufacture and enabled positioning in a wet environment. It allowed the application of total internal reflection fluorescence (TIRF) microscopy at the dorsal cell membrane, which is not adhering to a support. Specifically, the chip enabled TIRF microscopy at the apical membrane of polarized epithelial cells. Thereby, the device allowed us for the first time to monitor individual fusion events of GPI-GFP bearing vesicles at the apical membrane in live Madin-Darby canine kidney II (MDCK II) cells. Moreover, a mapping of fusion sites became feasible and revealed that the whole apical membrane is fusion competent. In total, the biochip offers an all-in-one solution for apical TIRF microscopy and contributes a novel tool to study trafficking processes close to the apical plasma membrane in polarized epithelial cells.     

A theoretical study of radical-only and combined radical/carbocationic mechanisms of arachidonic acid cyclooxygenation by prostaglandin H synthase

Prostaglandin H synthase catalyzes the oxygenation of arachidonic acid into the cyclic endoperoxide, prostaglandin G₂ (PGG₂), and the subsequent reduction of PGG₂ to the corresponding alcohol, prostaglandin H₂ (PGH₂), the precursor of all prostaglandins and thromboxanes. Both radical abstraction by a neighboring tyrosyl radical and combined radical/carbocationic models have been proposed to explain the cyclooxygenase part of this reaction. We have used density functional theory calculations to study the mechanism of the formation of the cyclooxygenated product PGG₂. We found an activation free energy for the initial hydrogen abstraction by the tyrosine radical of 15.6 kcal/mol, and of 14.5 kcal/mol for peroxo bridge formation, in remarkable agreement with the experimental value of 15.0 kcal/mol. Subsequent steps of the radical-based mechanism were found to happen with smaller barriers. A combined radical/carbocation mechanism proceeding through a sigmatropic hydrogen shift was ruled out, owing to its much larger activation free energy of 36.5 kcal/mol. Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00214-003-0476-9. Electronic Supplementary MaterialSupplementary material is available in the online version of this article at Electronic Supplementary Material: Supplementary material is available in the online version of this article at     

Video enhanced differential interference contrast microscopy: Characterizing colloidal materials

Video enhanced differential interference contrast microscopy (VEDICM) permits an immediate, rapid characterization of association colloid aggregates and other colloidal aggregates by direct visualization on a television screen. Particles with sizes down to 500 A, their dynamics, fusion and slow flocculation can be directly pictured, recorded and analyzed in real time, freezeframe, slow motion or time lapse. It is precisely in the distance regime, 500–10,000 A, joining micellar chemistry to the field of biological structures, that classical techniques do have most difficulty. In this domain surfactant aggregates-vesicles, liposomes, microemulsions, microtubules-can exhibit an astonishing dynamic structural diversity and distribution of structures. These are highly sensitive to pH, salt, temperature, and surfactant concentration in ways which are partially understood at a theoretical level, but not formerly easily accessible.In this paper, the VEDICM technique is described and its ability to follow the spontaneous growth and fusion of vesicles upon changes in chemical environment is presented.Session lecture, Ninth International Conference on Non-Aqueous Solutions, Pittsburgh, PA, August 1984.     

Measurement of coefficient of thermal expansion of films using digital image correlation method

Applications of the digital image correlation method (DIC) for the determination the coefficient of thermal expansion (CTE) of films is investigated in this paper. A heating chamber was designed for applying thermal load and DIC provides the full-field thermal deformation fields of the test film sample due to temperature changes. The average normal strains in the x and y direction from the region of interest are then extracted for the determination of CTE. The influence of unavoidable small rigid body rotation is discussed and a method to eliminate it to show the pure thermal expansion of the test film is demonstrated. For validation, the CTE of a pure copper sample is determined and compared with the textbook value, confirming the effectiveness and accuracy of the proposed technique. Finally, the CTE of Polyimide (PI) composite film in the temperature range of 20–140 °C is measured. The results reveal that the DIC is a practical and effective tool for full-field thermal deformation and CTE measurement of films.