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Super-resolution fluorescence microscopy can be achieved by image reconstruction after spatially patterned illumination or sequential photo-switching and read-out. Reconstruction algorithms and microscope performance are typically tested using simulated image data, due to a lack of strategies to pattern complex fluorescent patterns with nanoscale dimension control. Here, we report direct electron-beam patterning of fluorescence nanopatterns as calibration standards for super-resolution fluorescence. Patterned regions are identified with both electron microscopy and fluorescence labelling of choice, allowing precise correlation of predefined pattern dimensions, a posteriori obtained electron images, and reconstructed super-resolution images.
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Electrones , Iluminación , Calibración , Microscopía Fluorescente , AlgoritmosRESUMEN
High resolution dense lines patterned by focused electron beam-induced deposition (FEBID) have been demonstrated to be promising for lithography. One of the challenges is the presence of interconnecting material, which is often carbonaceous, between the lines as a result of the Gaussian line profile. We demonstrate the use of focused electron beam-induced etching (FEBIE) as a scanning electron microscope (SEM)-based direct-write technique for the removal of this interconnecting material, which can be implemented without removing the sample from the SEM for post processing. Secondary electron (SE) imaging has been used to monitor the FEBIE process, and atomic force microscopy (AFM) measurements confirm the fabrication of well separated FEBID lines. We further demonstrate the application of this technique for removing interconnecting material in high resolution dense lines using backscattered electron (BSE) imaging to monitor the process.
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Nanostructures of platinum-carbon nanocomposite material have been formed by electron-beam induced deposition. These consist of nanodots and nanowires with a minimum size â¼20 nm, integrated within â¼100 nm nanogap n-type silicon-on-insulator transistor structures. The nanodot transistors use â¼20 nm Pt/C nanodots, tunnel-coupled to Pt/C nanowire electrodes, bridging the Si nanogaps. Room-temperature single-electron transistor operation has been measured, and single-electron current oscillations and 'Coulomb diamonds' observed. In nanowire transistors, the temperature dependence from 290 to 8 K suggests that the current is a combination of thermally activated and tunnelling transport of carriers across potential barriers along the current path, and that the Pt/C is p-type at low temperature.
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The current understanding in the study of focused electron beam induced processing (FEBIP) is that the growth of a deposit is mainly the result of secondary electrons (SEs). This suggests that the growth rate for FEBIP is affected by the SE emission from the support. Our experiments, with membranes thinner than the SE escape depth, confirm this hypothesis. We used membranes of 1.4 and 4.3 nm amorphous carbon as supports. At the very early stage, the growth is support-dominated and the growth rate on a 4.3 nm thick membrane is three times higher than on a 1.4 nm thick membrane. This is consistent with Monte Carlo simulations for SE emission. The results suggest that SEs are dominant in the dissociation of W(CO)6 on thin membranes. The best agreement between simulations and experiment is obtained for SEs with energies between 3 and 6 eV.With this work we revisit earlier experiments, working at a precursor pressure 20 times lower than previously. Then, despite using membranes thinner than the SE escape depth, we did not see an effect on the experimental growth rate. We explain our current results by the fact that very early in the process, the growth becomes dominated by the growing deposit itself.
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Scanning ion microscopy has received a boost in the last decade, thanks to the development of novel ion sources employing light ions, like He(+), or ions from inert gases, like Ne(+) and Ar(+). Scanning ion images, however, might not be as easy to interpret as SEM micrographs. The contrast mechanisms are different, and there is always a certain degree of sample sputtering. The latter effect, on the one hand, prevents assessing the resolution on the basis of a single image, and, on the other hand, limits the probing time and thus the signal-to-noise ratio that can be obtained. In order to fully simulate what happens when energetic ions impact on a sample, a Monte Carlo approach is often used. In this paper, a different approach is proposed. The contrast is simulated using curves of secondary electron yields versus the incidence angle of the beam, while the surface modification prediction is based on similar curves for the sputtering yield. Finally, Poisson noise from primary ions and secondary electrons is added to the image. It is shown that the evaluation of an ion imaging tool cannot be condensed in a single number, like the spot size or the edge steepness, but must be based on a more complex analysis taking into account at least three parameters: sputtering, contrast and signal-to-noise ratio. It is also pointed out that noise contributions from the detector cannot be neglected for they can actually be the limiting factor in imaging with focused ion beams. While providing already good agreement with experimental data in some imaging aspects, the proposed approach is highly modular. Further effects, like edge enhancement and detection, can be added separately.
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It is often suggested that the growth in focused electron beam induced processing (FEBIP) is caused not only by primary electrons, but also (and even predominantly) by secondary electrons (SEs). If that is true, the growth rate for FEBIP can be changed by modifying the SE yield. Results from our Monte Carlo simulations show that the SE yield changes strongly with substrate thickness for thicknesses below the SE escape depth. However, our experimental results show that the growth rate is independent of the substrate thickness. Deposits with an average size of about 3 nm were written on 1 and 9 nm thick carbon substrates. The apparent contradiction between simulation and experiment is explained by simulating the SE emission from a carbon substrate with platinum deposits on the surface. It appears that the SE emission is dominated by the deposits rather than the carbon substrate, even for deposits as small as 0.32 nm(3).
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The low-voltage foil corrector is a novel type of foil aberration corrector that can correct for both the spherical and chromatic aberration simultaneously. In order to give a realistic example of the capabilities of this corrector, a design for a low-voltage scanning electron microscope with the low-voltage foil corrector is presented. A fully electrostatic column has been designed and characterised by using aberration integrals and ray tracing calculations. The amount of aberration correction can be adjusted relatively easy. The third order spherical and the first order chromatic aberration can be completely cancelled. In the zero current limit, a FW50 probe size of 1.0 nm at 1 kV can be obtained. This probe size is mainly limited by diffraction and by the fifth order spherical aberration.
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The creation of functional nanostructures by electron-beam-induced deposition (EBID) is becoming more widespread. The benefits of the technology include fast 'point-and-shoot' creation of three-dimensional nanostructures at predefined locations directly within a scanning electron microscope. One significant drawback to date has been the low purity level of the deposition. This has two independent causes: (1) partial or incomplete decomposition of the precursor molecule and (2) contamination from the residual chamber gas. This frequently limits the functionality of the structure, hence it is desirable to improve the decomposition and prevent the inclusion of contaminants. In this contribution we review and compare for the first time all the techniques specifically aimed at purifying the as-deposited impure EBID structures. Despite incomplete and scattered data, we observe some general trends: application of heat (during or after deposition) is usually beneficial to some extent; working in a favorable residual gas (ultra-high vacuum set-ups or plasma cleaning the chamber) is highly recommended; gas mixing approaches are extremely variable and not always reproducible between research groups; and carbon-free precursors are promising but tend to result in oxygen being the contaminant species rather than carbon. Finally we highlight a few novel approaches.
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Electrones , Nanoestructuras/química , Nanotecnología/métodos , Platino (Metal)/químicaRESUMEN
In the past decade, the feature size in ultra large-scale integration (ULSI) has been continuously decreasing, leading to nanostructure fabrication. Nowadays, various lithographic techniques ranging from conventional methods (e.g. photolithography, x-rays) to unconventional ones (e.g. nanoimprint lithography, self-assembled monolayers) are used to create small features. Among all these, resist-based electron beam lithography (EBL) seems to be the most suitable technique when nanostructures are desired. The achievement of sub-20-nm structures using EBL is a very sensitive process determined by various factors, starting with the choice of resist material and ending with the development process. After a short introduction to nanolithography, a framework for the nanofabrication process is presented. To obtain finer patterns, improvements of the material properties of the resist are very important. The present review gives an overview of the best resolution obtained with several types of both organic and inorganic resists. For each resist, the advantages and disadvantages are presented. Although very small features (2-5 nm) have been obtained with PMMA and inorganic metal halides, for the former resist the low etch resistance and instability of the pattern, and for the latter the delicate handling of the samples and the difficulties encountered in the spinning session, prevent the wider use of these e-beam resists in nanostructure fabrication. A relatively new e-beam resist, hydrogen silsesquioxane (HSQ), is very suitable when aiming for sub-20-nm resolution. The changes that this resist undergoes before, during and after electron beam exposure are discussed and the influence of various parameters (e.g. pre-baking, exposure dose, writing strategy, development process) on the resolution is presented. In general, high resolution can be obtained using ultrathin resist layers and when the exposure is performed at high acceleration voltages. Usually, one of the properties of the resist material is improved to the detriment of another. It has been demonstrated that aging, baking at low temperature, immediate exposure after spin coating, the use of a weak developer and development at a low temperature increase the sensitivity but decrease the contrast. The surface roughness is more pronounced at low exposure doses (high sensitivity) and high baking temperatures. A delay between exposure and development seems to increase both contrast and the sensitivity of samples which are stored in a vacuum after exposure, compared to those stored in air. Due to its relative novelty, the capabilities of HSQ have not been completely explored, hence there is still room for improvement. Applications of this electron beam resist in lithographic techniques other than EBL are also discussed. Finally, conclusions and an outlook are presented.
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Electrones , Nanotecnología/métodos , Microscopía de Fuerza Atómica , Microscopía Electrónica de Rastreo , Nanocompuestos , Nanoestructuras/química , Compuestos de Organosilicio/química , Fotograbar , ImpresiónRESUMEN
An attempt has been made to reach the ultimate spatial resolution for electron beam-induced deposition (EBID) using W(CO)(6) as a precursor. The smallest dots that have been written have an average diameter of 0.72 nm at full width at half maximum (FWHM). A study of the nucleation stage revealed that the growth is different for each dot, despite identical growth conditions. The center of mass of each dot is not exactly on the position irradiated by the e-beam but on a random spot close to the irradiated spot. Also, the growth rate is not constant during deposition and the final deposited volume varies from dot to dot. The annular dark field signal was recorded during growth in the hope to find discrete steps in the signal which would be evidence of the one-by-one deposition of single molecules. Discrete steps were not observed, but by combining atomic force microscope measurements and a statistical analysis of the deposited volumes, it was possible to estimate the average volume of the units of which the deposits consist. It is concluded that the volume per unit is as small as 0.4 nm(3), less than twice the volume of a single W(CO)(6) molecule in the solid phase.
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Recently, the fabrication resolution in electron beam-induced deposition (EBID) has improved significantly. Dots with an average diameter of 1 nm have been made. These results were all obtained in transmission electron microscopes on thin samples. As one may think that such resolution can be achieved on thin samples only, it is the objective of this paper to show that this should also be possible on thick samples. For that purpose we use Monte Carlo simulations of the electron-sample interaction and determine the surface area where secondary electrons are emitted. Assuming that these electrons cause the deposition in EBID, a comparison can be made between deposition on a thin and a thick sample. The Monte Carlo code we developed will be described and applied to the deposition induced by a 200 keV primary electron beam on an ultra-thin (10 nm) and a bulk-like (1,000 nm) Cu sample. Near the point of incidence of the primary beam, the deposit size is independent of the substrate thickness, such that a 1-nm resolution should be possible to achieve on a thick substrate as well. Thicker substrates only affect the tails of the deposit distribution which contain more mass than thin substrate deposit tails.
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In this study, we demonstrated the use of electron-beam-induced deposition for synthesis of artificial two-dimensional crystals with an in situ scanning transmission electron microscope. The structures were deposited from W(CO)6 in an environmental scanning transmission electron microscope on a 30-nm-thick Si3N4 substrate. We present clear electron beam diffraction patterns taken from those structures. The distance between the diffraction peaks corresponded to the dot spacing in the self-made surface crystal. We propose using these arrays of dots as anchor points for making artificial crystals for diffraction analysis of weakly scattering or beam-sensitive molecules such as proteins.
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A spherical and chromatic aberration corrector for electron microscopes is proposed, consisting of a thin foil sandwiched between two apertures. The electrons are retarded at the foil to almost zero energy, so that they can travel ballistically through the foil. It is shown that such a low-voltage corrector has a negative spherical aberration for not too large distances between aperture and foil, as well as a negative chromatic aberration. For various distances the third- and fifth-order spherical aberration coefficients and the first- and second-order chromatic aberration coefficients are calculated using ray tracing. Provided that the foils have sufficient electron transmission the corrector is able to correct the third-order spherical aberration and the first-order chromatic aberration of a typical low-voltage scanning electron microscope. Preliminary results show that the fifth-order spherical aberration and the second-order chromatic aberration can be kept sufficiently low.
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Extracts of the flower petals of Impatiens balsamina L. contain enzymes which catalyze the glycosylation of phenolic compounds. Enzymes have been extracted which glycosylate hydroquinone to arbutin and at least 3 different flavonols to the 3-monoglucoside. The hydroquinone glucosylating enzyme is similar to enzymes previously described except that it requires an unidentified low molecular weight cofactor. The glucosylation of flavonols follows normal enzyme kinetics; it requires a nucleotide diphosphate glucose donor for activity, and is made more evident by the presence of glucono-1:5-lactone, an inhibitor of endogenous glucosidases. It is suggested that the flavonol glucosylating enzyme acts naturally to glucosylate a precursor of both flavonols and anthocyanins to the 3-monoglucoside. The only elaboration of an anthocyanin observed with petal extracts was an acylation of pelargonidin-3-monoglucoside.