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Nanofluidic and monolithic environmental cells for cryogenic microscopy
摘要： We present a device capable of combining nanofluidics and cryogenic transmission electron microscopy (cryo-TEM) to allow inspection of water-soluble samples under near-native conditions. The devices can be produced in a multitude of designs, but as a general rule, they consist of channels or chambers enclosed between two electron-transparent silicon nitride windows. With the appropriate design, those devices can allow screening of multiple samples in parallel and remove the interaction between the sample and the environment (no air–water interface). We demonstrate channel sizes from 80 to 500 nm in height and widths from 100 to 2000 μm. The presented fabrication flow allows producing hollow devices on a single wafer eliminating the need of aligning or bonding two half-cavities from separate wafers, which provides additional resistance to thermal stress. Taking advantage of a single-step through-membrane exposure with a 100 keV electron beam, we introduced arrays of thin (10–15 nm) electron-transparent silicon nitride membrane windows aligned between top and bottom (200–250 nm) carrier membranes. Importantly, the final devices are compatible with standard TEM holders. Furthermore, they are compatible with rapid freezing of samples, which is crucial for the formation of vitreous water, hence avoiding the formation of crystalline ice, that is detrimental for TEM imaging. To demonstrate the potential of this technology, we tested those devices in imaging experiments verifying their applicability for cryo-TEM applications and proved that vitreous water could be prepared through conventional plunge freezing of the chips.
关键词： nanofabrication,TEM,environmental chamber,microfluidic cell,electron beam lithography,cryo-TEM
更新于2025-09-23 15:23:52
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Stereochemistry controlled supramolecular architectures of novel tetrahydroxy functionalized amphiphilic carbocyanine dye
摘要： The synthesis of novel amphiphilic 5,5',6,6'-tetrachlorobenzimidacarbocyanine (TBC) dye derivatives with amino-propanediol head groups differing in stereochemistry (chiral enantiomers, meso-form, and conformer) is reported. For the achiral meso-form, a new synthetic route towards asymmetric cyanine dyes was established. All compounds form J-aggregates in water whose optical properties were characterized by spectroscopic methods. The supramolecular structure of the aggregates was investigated by cryo-transmission electron microscopy (cryo-TEM), cryo-electron tomography (cryo-ET), and atomic force microscopy (AFM) revealing extended sheet-like aggregates for the chiral enantiomers and nanotubes for the mesomer, respectively, while the conformer forms predominately needle-like crystals. The experiments demonstrate that the aggregation behavior of compounds can be controlled solely by head group stereochemistry which in case of the enantiomers enables formation of extended hydrogen bond chains by the hydroxyl functionalities. In case of the achiral meso-form, however, such chains turned out to be sterically excluded.
关键词： Stereochemistry,cryo-TEM,supramolecular architectures,amphiphilic carbocyanine dye,cryo-ET,J-aggregates,AFM
更新于2025-09-19 17:13:59
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Synthesis and Characterization of Thiolate-Protected Gold Nanoparticles of Controlled Diameter
摘要： Gold nanoparticles (AuNPs) have been the focus of many studies owing to their unique optical and electronic properties and versatile applications. However, synthesis of stable and homogeneous AuNPs with a particular choice of size is still a challenge. In this study we describe a direct synthesis approach to produce stable and monodisperse water-soluble AuNPs with a tightly controlled diameter in the 1.7?2.4 nm range. We controlled the size by changing only the sodium hydroxide (NaOH) concentration in the synthesis. Gel electrophoresis, transmission electron microscopy (TEM), and solution X-ray scattering showed that the AuNPs had narrow size-distributions. We further showed that AuNPs of the di?erent sizes were clearly distinguishable in TEM micrographs, paving the way to dual-target labeling. The reactivity of the AuNPs toward DNA and proteins was also demonstrated. We utilized this reactivity to label tail-anchored proteins embedded in the membrane of the anticancer nanodrug Doxil as a means to target it to speci?c cell types. The gold-labeling enabled the precise localization of the tail-anchored proteins in cryo-TEM images of the therapeutic liposomes.
关键词： thiolate-protected,bioconjugation,cryo-TEM,tail-anchored proteins,synthesis,size control,Gold nanoparticles,Doxil
更新于2025-09-16 10:30:52