Scn5a mRNA. Overexpression of Nup107 induced the protein expression of Scn5a rather than that of other ion channels, with no effects of their mRNA levels. The analysis for the protein production demonstrated Nup107‐facilitated transport of Scn5a mRNA. Using RIP‐PCR and luciferase assay, we found that the 5′‐UTR of Scn5a mRNA was not involved in the interaction, whereas the spatial interaction between Nup107 protein and Scn5a mRNA was formed when Scn5a mRNA passing through the nuclear pore. Functionally, Nup107 overexpression in neonatal rat ventricle myocytes significantly increased the currents of Scn5a‐encoded INa channel. Moreover, the close correlation between Nup107 and Nav1.5 protein expression was observed in cardiomycytes and heart tissues subjected to hypoxia and ischaemic insults, suggesting a fast regulation of Nup107 on Nav1.5 channel in cardiac myocytes in a posttranscriptional manner. These findings may provide insights into the emergent control of cardiac electrophysiology through Nup‐mediated modulation of ion channels.Nucleoporins (Nups) are known to be functional in nucleo‐cytoplasmic transport, but the roles of nucleoporins in nonproliferating cells, such as cardiac myocytes, are still poorly understood. In this study, we report that Nup107 regulates cardiac bioelectricity by controlling the nucleo‐cytoplasmic trafficking of  The spatial interaction between Nup107 protein and Scn5a mRNA is implicated in the regulation of Scn5a mRNA export. Furthermore, we showed that Nup107 overexpression was functional in increasing the amplitude of Scn5a‐encoded Na+ currents, and associated with ischaemic cardiac injury by increasing the nucleo‐cytoplasmic transport of Scn5a mRNA.In this study, we demonstrated that Nup107 regulates the cardiac electricity by facilitating 22.1Animals in the study were maintained in accordance with the Guide for the Care and Use of Laboratory Animals (NIH Publication). All the experimental procedures with animals used in this study were approved by the Institutional Animal Care and Use Committee of Tongji University School of Medicine.2.2Myocardial infarction (MI) was performed in female Sprague Dawley rats as described previously.2.3−1 BrdU for 24 hours and maintained in DMEM containing 10% FBS, 2 mmol L−1 L‐glutamine and 1% penicillin/streptomycin (P/S) . HEK293 cells were cultured and maintained in DMEM with 10% FBS and 1% P/S.Neonatal rat ventricle myocytes (NRVMs) were isolated from hearts of 1‐ to 3‐day‐old Sprague Dawley rats. Cardiomyocytes were cultured in Dulbecco's modified Eagle's medium (DMEM) medium containing 10% foetal bovine serum (FBS) and 100 μmol L2.4® II One Step Cloning Kit . Different truncates of Nup107 were constructed, including Nup107 full length (FL), Nup107‐N‐terminus (1‐304 aa), Nup107‐conserved domain , Nup107‐C‐terminus (664‐925 aa), Nup107 truncates lacking the N‐terminus (ΔN‐terminus), lacking the conserved domain (ΔCD), or lacking the C‐terminus (ΔC‐terminus), by amplifying the corresponding sequences. The nucleotide sequences of these constructs were confirmed by Sanger sequencing. Transient plasmid transfections were performed with Lipofectamine 3000 (Invitrogen) according to the manufacturer's protocol.The full‐length of human Nup107 was PCR‐amplified from the cDNA of HEK293 cells and cloned into pCMV3‐GFPSpark  using ClonExpress2.5Recombinant adenoviruses expressing green fluorescent protein (Ad‐GFP) and Nup107 (Ad‐Nup107) were prepared using an AdEasy vector system . NRVMs were isolated, cultured, and infected with Ad‐Nup107 and Ad‐GFP adenoviruses (multiplicity of infection = 100).2.6Scn5a mRNAs were calculated using the DeltaDeltaCT method, and were normalized against the levels of multiple housekeeping genes . All measurements were performed in triplicate. Melting curve analysis confirmed amplification specificity.Total RNA was extracted using Trizol reagent (Invitrogen) and reverse transcribed into cDNA using the PrimeScript RT reagent Kit . Detection of gene expression was performed with SYBR Green Master Mix and the 7900 Real‐Time PCR System . The expression levels of For the detection of the mRNA levels in different fractions, RNAs from the cytoplasm and nucleus were isolated according to the manufacturer's instructions . Each RNA fraction was subsequently reverse‐transcribed using PrimeScript RT reagent Kit . The absolute quantitative method was used to estimate the relative amount of indicated mRNA in cytoplasm and nucleus. This method is based on the construction of a standard curve of cycle number at a threshold (CT) vs the initial input amount of mRNA copy number. The PCR products containing fragments of the cDNA of each gene were used as the initial input. All the primer sequences are listed in Table 2.7Total proteins were extracted with RIPA buffer  containing protease inhibitor cocktail tablets , and separated by SDS‐PAGE (Invitrogen). Then they were transferred onto PVDF membranes and probed with antibodies against Nav1.5, Kv4.3, and Cav1.2 , Kir2.1 and Kv1.4 , Nup107 and GFP , β‐actin, and GAPDH . After washing, blots were treated with the appropriate IRDye 800 conjugated secondary antibody  at room temperature for 1 hours. Images were recorded using the Odyssey infrared imaging system and analysed using the Odyssey Application Software v2 (LI‐COR Biosciences).2.8Scn5a mRNA, the luciferase reporter vectors harbouring different parts of Scn5a mRNA (5′‐untranslated region (UTR), coding sequence (CDS), and 3′‐UTR) were constructed as follows: the sequence of 5′‐UTR of rat Scn5a mRNA (NM_013125) was synthesized and inserted into the 5′ NHE I of the Renilla luciferase gene in psiCheck2 . The different regions of the 3′‐UTR of Scn5a mRNA  were amplified from the cDNA of H9c2 cells and cloned into the 3′ downstream of the Renilla luciferase gene using the Xho I and Not I sites. The CDS regions of human SCN5A mRNA  were amplified from pCMV6‐SCN5A‐GFP  and cloned into the Xho I and Not I sites of psiCheck2 vector. The resultant psiCheck2 constructs were cotransfected with Nup107 and its truncates, or control vector in HEK293 cells for 48 hours. Afterwards the firefly and renilla luciferase activities were analysed using the Dual‐Luciferase Reporter Assay System (Promega).For the functional analysis of 2.9Scn5a mRNA immunoprecipitation. Antibodies used for RIP included rabbit polyclonal IgG , GFP, and Nup107 , and 5 μg of antibody was used per RIP reaction. The immunoprecipitated RNA was extracted with Trizol and reverse‐transcribed with PrimeScript RT reagent Kit (Takara). All RIP assays were performed with biological duplicates. The RIP‐PCR was used to measure the amount of Scn5a mRNA in pull‐down samples. The data were demonstrated as the percentage of input GAPDH mRNA of its respective group. P values were obtained using two‐tailed Student's t‐test.The RIP experiment was performed using a Millipore EZ‐Magna RIP RNA‐Binding Protein Immunoprecipitation kit  according to the manufacturer's instructions. NRVMs were infected with Ad‐Nup107 or Ad‐GFP for 48 hours and harvested for the endogenous 2.10Na was recorded with NRVMs bathed in a solution containing (in mmol L−1): 20 NaCl, 50 TEA‐Cl, 67 CsCl, 1 MgCl2, 1 CaCl2, 0.1 CdCl2, 10 glucose and 10 Hepes, pH 7.4 with CsOH. Pipette solution consisted of (in mol L−1) 5 NaCl, 125 CsF, 10 EGTA, 10 Hepes and 5 Mg‐ATP,pH 7.2 with CsOH. Cells were held at −120 mV and INa was elicited by a family of voltage steps to potentials ranging from −70 to +40 mV with 10 mV increments for 50 ms, with 5000 ms interpulse intervals.Whole‐cell patch‐clamp recordings were performed at room temperature (24°C) using an EPC‐10 amplifier and pulse software  as previous reports.2.11t‐test, as appropriate. The results were considered statistically significant if P < 0.05.All data are presented as the mean ± SD. Statistical comparisons among multiple groups were performed by ANOVA and unpaired Student's 33.1Scn5a‐encoded Nav1.5 protein, whereas other ionic channels, such as calcium channel (Cacna1c) and potassium channels  were not affected in both mRNA and protein expression .3.2Scn5a mRNA. We designed the primers that annealed to the intron (int), or the junction region spanning the junction (junc) of the intron and exon to detect the unspliced Scn5a mRNA, or the exon‐spanning primers that detected only the spliced, mature Scn5a mRNA. To exclude the possibility that the overexpression of Nup107 could affect the expression of internal references, multiple genes, such as GAPDH, β‐actin, β‐tubulin, and 18s, were used to normalize the mRNA level of Scn5a mRNA. Our data showed that Ad‐Nup107 infection did not affect the production of either nascent or mature Scn5a mRNA in NRVMs, compared with the Ad‐GFP‐treated cells  did not alter the lifespan of Nav1.5 protein in Nup107‐overexpressed NRVMs , coding DNA sequence (CDS), and 3′‐UTR, were used to identify the potential regions interactive with Nup107 , an Nup84p‐homologous conserved domain (CD) (aa 305‐664) and a C‐terminal region (aa 665‐925). To identify the critical domain responsible for d Figure  and coex04, an Nu3.4−1, n = 20), compared with the control Ad‐GFP group . These data indicate that Nup107 overexpression can increase the amplitude of Nav1.5 channel currents in cardiac myocytes.To determine whether Nup107 could affect the function of Nav1.5 channel, we did the whole‐cell patch‐clamping recordings for the Nup107‐mediated electrophysiological regulation. Our data demonstrated that the current density of the voltage‐gated Nav1.5 channel was dramatically increased in adenovirus‐mediated overexpression of Nup107 in NRVMs Figure A and B. 2 treatment  resulted in marked increase in Nup107 and Nav1.5 proteins in NRVMs (Figure Scn5a may also be relevant to ischaemic cardiac diseases in vivo.We next explored the potential contribution of Nup107‐related Nav1.5 expression in pathological heart tissues. Following the development of hypoxia, the protein levels of Nup107 and Nav1.5 were both upregulated at 24 hours, and gradually downregulated at 48 hours after 1% Os Figure . These d4Scn5a mRNA nuclear export and Scn5a‐encoded Nav1.5 channel in cardiomyocytes. First, Nup107 specifically regulates the distribution of Scn5a mRNA in a posttranscriptional manner. Second, active interaction between Nup107 protein and Scn5a mRNA is required for the Scn5a mRNA export and subsequent translation. Third, Nup107 increment is functional in regulating cardiac electricity in that its overexpression increases the current densities of Scn5a‐encoded Nav1.5 channel. Fourth, Nup107‐mediated regulation of Scn5a‐encoded Nav1.5 channel is implicated in hypoxia‐induced myocytes and ischaemic cardiac tissues. These findings suggest that the Nup107‐dependent export of Scn5a mRNA is a critical step in regulating cardiac electrical activity.This study provides multiple lines of evidence implicating the role of Nup107 in regulating The Nup107 complex is known to be a pivotal player in NPC formation. It is located in the central scaffold of the NPC and mediates mRNA export in interphase, and has roles in kinetochore function, spindle assembly, and postmitotic nuclear pore assembly. Data mining Scn5a, but not other ion channel genes, such as calcium and potassium channels. The regulation occurred in the transportation stage, in which the mature Scn5a mRNA was exported from nucleus to cytoplasm. This is consistent with the localization of Nup107 in the NE,Our data showed that Nup107 specifically regulated the expression of Scn5a mRNA is formed before its entry into the cytoplasm. Interestingly, the 5′‐UTR, which is demonstrated to be a crucial part, guiding the mRNA through the nuclear pore,Scn5a mRNA are all involved in the contact with Nup107 (Figure Scn5a mRNA export. As one of the largest mRNAs, the formation of this conformational Scn5a mRNA may be necessary and a rate‐limiting step for mRNA export.Till now we do not know the detail of this posttranscriptional regulation, but it seems that the three‐dimensional shape of 7 Figure D. No defScn5a mRNA interaction (Figure Scn5a mRNA. Given that Scn5a mRNA forms a three‐dimensional structure before its nuclear export, it is understandable that this conformation of Scn5a mRNA is induced by the flexible Nup107, and consequently the interaction between Nup107 protein and Scn5a mRNA is dynamically changed in the process of mRNA passing through the nuclear pore. We do not know exactly whether the FG repeats play a role in this reciprocal action, despite Nup107 contains several FG repeats, but the truncated mutant of Nup107 lacking the FG repeats exhibits the similar results to the full‐length of Nup107, indicating the dispensable role of FG repeats in the translocation of Scn5a mRNA.Mammalian Nup107 is composed of an N‐terminal region, a C‐terminal region and an Nup84p‐like conserved domain, which is the homologous domain of Nup84p in yeast. Unexpectedly no special domain of Nup107 was found to be critical for the Nup107 protein‐n Figure E and F. Scn5a mRNA is regulated indirectly by other nucleoporins, for example, by a complex containing multiple types of nucleoporins.Scn5a mRNA (Figure It has been reported that Nup107 forms the Nup107‐160 subcomplex with a number of other nucleoporinsA Figure , and theScn5a gene, is an integral membrane protein. As the pore forming ɑ‐subunit of the voltage‐dependent cardiac Na+ channel, it is fundamental in the initiation and conduction of action potentials in cardiac myocytes.in vitro and in vivo underlies the regulation of Scn5a mRNA by Nup107 under pathological condition. Importantly, the increased expression of Nup107 was observed both in hypoxia and oxidative stress in cardiomyocytes, suggesting that Nup107 is a fast reactive protein responding to the insults. This may represent an emergent responsive mechanism, when the cardiac myocytes encounter the impaired environment.Scn5a mRNA. This is supported by the observation of MI model showing the increased Nup107 and Nav1.5 expression at 72 hours, and increasing Nup107 protein and decreased Nav1.5 expression at 1 week after MI (Figure Nav1.5, encoded by I Figure . The 72 In summary, our findings clarify for the first time the mechanism by which Nup107 regulates cardiac electrical activity through controlling posttranscriptional transportation, and has provided valuable insight into the molecular modulator in cardiac diseases.The authors confirm that there are no conflicts of interest. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file. Click here for additional data file.


The excellent photo and temporal responses, which generally show a trade‐off relation, are achieved simultaneously by applying a p‐doping technique based on hydrochloric acid (HCl) to a selected ReSe2 region. Because the p‐doping of ReSe2 originates from the charge transfer from un‐ionized Cl molecules in the HCl to the ReSe2 surface, by adjusting the concentration of the HCl solution from 0.1 to 10 m, the doping concentration of the ReSe2 is controlled between 3.64 × 1010 and 3.61 × 1011 cm−2. Especially, the application of the selective HCl doping technique to the ReSe2 photodetector increases the photoresponsivity from 79.99 to 1.93 × 103 A W−1, and it also enhances the rise and decay times from 10.5 to 1.4 ms and from 291 to 3.1 ms, respectively, compared with the undoped ReSe2 device. The proposed selective p‐doping technique and its fundamental analysis will provide a scientific foundation for implementing high‐performance TMD‐based electronic and optoelectronic devices.In this study, a near‐infrared photodetector featuring a high photoresponsivity and a short photoresponse time is demonstrated, which is fabricated on rhenium diselenide (ReSe 2) photodetector featuring long wavelength detection, short response time, and high photoresponsivityA rhenium diselenide (ReSe is demonstrated by applying a selective HCl p‐doping technique to the ReSe2 channel region. The effects of the HCl p‐doping on the ReSe2 material and device are thoroughly investigated via Raman spectroscopy, Kelvin probe force microscopy, and electrical measurements in dark and under light illumination conditions. Lopez‐Sanchez et al.2 photodetectors with high photoresponsivity (880 A W−1 under 550 nm and 0.12 A W−1 under 633 nm) and low dark currents (2 pAVG = −70 V and 10 pAVG = −1.2 V). However, these photodetectors based on group VI TMDs were unable to detect infrared light (λ > 900 nm) owing to their wide energy bandgap of >1.2 eV.2), which has a relatively narrow bandgap (0.9–1.0 eV),2 devices exhibited poor photoresponsivity compared with the group VI TMD photodetectors. The maximum photoresponsivity values of O2 plasma‐treated and Mo‐doped ReSe2 photodetectors were 95−1,3)2 photodetectors, we significantly improved their photoresponsivity to 4.2 × 104 and 1.18 × 106 A W−1, respectively. However, the photoresponse time of these n‐doped devices was significantly degraded owing to the reduced depletion width at the metal/ReSe2 junction and the consequent reduction in photocarrier collection (τ: 2.7 ms → 4.5 ms after APTMS doping and τ: 64 ms → 263 ms after PPh3 doping).In the past decade, owing to the superior optical properties of transition‐metal dichalcogenides (TMDs), such as their excellent light–matter interaction,2 photodetector featuring long wavelength detection, short response time, and high photoresponsivity by applying a p‐doping process based on hydrochloric acid (HCl) to a selected ReSe2 region. The built‐in potential at the p+/p junction formed by the HCl p‐doping allowed the ReSe2 photodetector to have a high photoresponsivity and a short photoresponse time simultaneously. The effects of the HCl p‐doping on the ReSe2 material and device were thoroughly investigated via Raman spectroscopy, Kelvin probe force microscopy (KPFM), and electrical measurements in the dark and under light illumination.Here, we demonstrated a ReSe22 crystal material through microscopic analyses, such as Raman spectroscopy and KPFM. Figure2 material before and after surface treatment with 10 m HCl. Several peaks were observed in the spectral range of 100–300 cm−1. This is because the distorted triclinic structure of ReSe2 caused 18 Raman‐active vibration modes,2, WSe2, MoSe2, and WS2).m HCl was applied to the ReSe2, all the Raman peaks were slightly blue‐shifted (gray solid line → red dotted line). This is likely because the in‐ and out‐of‐plane vibrations of the ReSe2 structure were intensified by the HCl treatment.2 by octadecyltrichlorosilane (OTS) caused the (E12g + A2g) peak to blue‐shift by −2.03 cm−1.g‐like and Ag‐like peaks of ReSe2 after doping with various HCl concentrations . We prepared three samples for each HCl concentration and randomly selected five points on each sample for the Raman measurements. Thus, the data points and error bars in Figure m, the peak‐shift values increased from 0.14 to 0.93 cm−1 for the Eg‐like peaks and from 0.24 to 1.1 cm−1 for the Ag‐like peaks. To confirm the p‐doping effect on ReSe2 once again, we performed KPFM on ReSe2 crystal flakes before and after doping with 10 m HCl, as shown in Figure m HCl solution. For quantitative analysis, we extracted the average work‐function values (ΦReSe2) from the KPFM mapping images and plotted them, as shown in Figure ΦReSe2 increased by 0.13 eV (from 4.48 to 4.61 eV) after doping with the 10 m HCl solution, and the Fermi‐level shift toward the valence band indicated the p‐type doping effect of HCl on the ReSe2. Furthermore, as indicated by the histograms of ΦReSe2 shown in Figure ΦReSe2 values were uniformly distributed near 4.48 and 4.61 eV before and after the 10 m HCl doping, respectively (80% of the pixels were in the range of ±0.3 eV).First, we investigated the effect of HCl treatment on the ReSe2 transistors and performed electrical measurements on the devices before and after HCl doping to investigate the effects of the doping on the transistor performance. For reliable analysis, we prepared three different ReSe2 devices for each HCl doping condition and presented error bars for each data point. Figure2 transistor and the energy band diagrams of the Pt/ReSe2/Pt region before and after the HCl doping. The HCl doping is expected to be induced by the surface charge‐transfer phenomenon between the un‐ionized Cl molecules and the ReSe2 layer. The un‐ionized Cl molecules have a higher molecular electron affinity than the ReSe22. Further experimental proof about the HCl p‐doping based on the Cl molecules is provided in Figure S7 in the Supporting Information. Owing to this electron‐transfer phenomenon, a change in the Fermi level of ReSe2 was observed after the HCl doping [4.48 eV → 4.61 eV in the case of 10 m HCl, as shown in Figure Φeff_hole) at the source Pt/ReSe2 junction and increased the hole‐injection probability from the source Pt to the ReSe2. This variation in Φeff_hole affected the on‐current (Ion) and threshold voltage (VTH) of the ReSe2 transistor. As shown in Figure m HCl, Ion increased from 27.8 to 97.7 nA µm−1 , and VTH was shifted from −8.21 to 7.21 V. The controllability of the HCl doping on the ReSe2 transistor was then examined with regard to the threshold‐voltage shift (ΔVTH = VTH_HCl − VTH_Control) and the carrier concentration (Δp = Δp_HCl − Δp_Control), as shown in Figure m. The Δp was extracted from the ID–VG curves using the following equation: p = IDL/qWµVD, where L and W represent the channel length and width, respectively, q represents the electron charge, and µ represents the mobility of the ReSe2 device. As the HCl concentration increased from 0.1 to 10 m, the ΔVTH and Δp changed from 3.06 to 15.41 V and from 3.64 × 1010 to 3.61 × 1011 cm−2, respectively. These p‐doping concentrations were comparable to the reported values for p‐doping techniques: from 2.1 × 1011 to 5.2 × 1011 cm−2 (OTS on WSe2),12 cm−2 (FOTS doping on MoS2),10 cm−2 (M‐DNA on MoS2 and WSe2),12 cm−2 (O2 plasma treatment on ReS2),12 cm−2 (NO2 doping on WSe2),11 to 1.32 × 1012 cm−2 (HCl doping on WSe2).ION and µFE after/before the p‐doping increased from 2.05 to 3.52 and from 2.55 to 4.37, respectively . Electrical measurements were performed again in the dark and under laser illumination. As shown in Figure 2 channel region. The fully doped ReSe2 device exhibited a higher photocurrent than the control ReSe2 device in the entire gate‐voltage region. Here, “fully doped ReSe2” indicates that the whole opened area of ReSe2 was doped  at the boundary of the pristine and HCl‐doped regions. This indicates that the internal electric field was formed in the boundary region of 0.65 µm owing to the difference in the carrier concentration. A schematic of the selectively doped Pt/ReSe2/Pt device structure with a back‐gate electrode is presented in Figure 2 as the source and the electrode on HCl‐doped ReSe2 as the drain. Figure ID–VD characteristics of the control and selectively doped ReSe2 devices, where VGS is 0 V. In the control device, similar current levels were observed in the negative and positive drain voltage regions. In the selectively doped ReSe2 device, a higher current was observed in the positive‐voltage region (32.7 nA µm−1 at VDS = 5 V and 1.26 nA µm−1 at VDS = −5 V). This is because the carrier injection at the ReSe2/Pt drain junction was enhanced by the selective doping. Figure 2/Pt junctions formed on the selectively doped ReSe2, which were drawn under different drain bias conditions . In the case of a negative drain voltage, because high barriers were initially formed at the both Pt/ReSe2 (source side) and ReSe2/Pt (drain side) junctions, no reduction in the drain current was observed, even though an increase in the effective hole barrier height was predicted due to the selective doping. In contrast, under the positive drain bias condition, the hole‐carrier injection at the Pt/WSe2 drain junction was expected to increase significantly owing to the selective doping of ReSe2. This manifested as an increase in the drain current by more than two orders of magnitude. Additionally, we provide the energy band diagrams of the control, fully doped, and selectively doped Pt/ReSe2/Pt junctions under different drain bias conditions in Figure S2 in the Supporting Information.To improve the photoresponsivity further, we then formed an internal electric field in the middle of the ReSe2 photodetectors was evaluated, as shown in Figure2. We applied lasers with power of 10 nW and wavelengths between 520 and 980 nm to the three types of devices and measured the photocurrents. The selectively doped ReSe2 device exhibited a higher photocurrent than the control ReSe2 device by at least one order of magnitude, and the photocurrent was higher than that of the fully doped ReSe2 device . Under laser illumination of λ = 520 nm, the photocurrent values (VGS = 0 V) for the control, fully doped, and selectively doped ReSe2 devices were 1.61 × 10−7, 6.32 × 10−7, and 3.95 × 10−6 A µm−1, respectively. We then obtained the photoresponsivity values at the bias point of VGS = 0 V and VDS = −5 V in the IPhoto versus VGS characteristic curves, which are plotted with respect to the wavelength in Figure 2 device, relatively high photoresponsivity values appeared between 1.93 × 103 (λ = 520 nm) and 208.8 (λ = 980 nm) A W−1. The photoresponsivity values of the control and fully doped devices were distributed between 79.99 and 10.29 and between 314.4 and 41.96, respectively, in the same wavelength range. Figure 2 device under two wavelength conditions, which was plotted with respect to the incident laser power. The highest photoresponsivity values under each wavelength condition were obtained at the lowest power of 5.02 pW: 9.1 × 103 and 4.96 × 103 A W−1 at λ = 520 and 980 nm, respectively. Overall, a higher photoresponsivity was achieved at a lower laser power. This inverse relationship between the photoresponsivity and the incident laser power was due to the suppression of the scattering of photocarriers under the low‐power condition.2 devices at VDS = −5 V and VGS = −30 V. As previously mentioned, by applying the HCl doping to the entire ReSe2 region, a higher photocurrent was obtained, but a higher dark current was observed owing to the increased hole‐injection probability. As shown in the rightmost diagram, the selective HCl doping applied to the right‐half region of ReSe2 improved the photocarrier collection by forming an internal electric field at the p+/p junction boundary and also maintained the hole‐carrier injection from the source. In addition, the defects on the ReSe2 surface were expected to be passivated by the HCl treatment. This appeared to reduce the recombination rate in ReSe2 and to increase photocarrier lifetime and photoresponsivity.Finally, the performance of control, fully doped, and selectively doped ReSeIPhoto/IDark ratios of the fully and selectively doped ReSe2 devices , and then the effective electron barrier height at the drain‐side ReSe2/Pt junction decreased (a dark current increased). The control, fully doped, and selectively doped ReSe2 devices exhibited maximum IPhoto/IDark values of 6.24 × 103, 4.31 × 104, and 1.75 × 105 at the gate voltages of 12.7, 20.8, and 28.1 V, respectively, where the minimum dark currents appeared. The IPhoto/IDark of the selectively doped device was superior to those of the other devices over the entire gate‐voltage region owing to the lower dark current and higher photocurrent of this device. Finally, the time‐dependent photoresponse characteristic was investigated for the three types of ReSe2 photodetectors. Figure r) and decay (τd) times at 20 different rising and decaying edges, which were plotted as shown in Figure r and τd) were 10.5 and 291 ms, respectively, for the control ReSe2 device; 17 and 323 ms, respectively, for the fully doped ReSe2 device; and 1.4 and 3.1 ms, respectively, for the selectively doped ReSe2 device. As previously mentioned, the selectively doped ReSe2 photodetector exhibited the fastest response to the laser pulses, and the response of the control ReSe2 device was slightly faster than that of the fully doped device. The selective doping formed an electric field at the interface between the doped and undoped ReSe2 regions, enhancing the splitting of the photogenerated electron–hole pairs. However, HCl doping of the entire ReSe2 region narrowed the depletion width at the Pt/ReSe2 junctions compared with the control device, hindering the photocarrier splitting and thereby increasing the photoresponse time.In addition to the photoresponsivity, the dark‐current level is important for evaluating the performance of photodetectors. Thus, we investigated the s Figure e. As sho32 photodetector for detecting 520−980 nm wavelength light by using an HCl‐based selective p‐doping technique . The effect of p‐type doping on the ReSe2 material was experimentally investigated via Raman spectroscopy and KPFM. Because the p‐doping of ReSe2 was based on the charge transfer phenomenon from the un‐ionized Cl molecules to the ReSe2 surface, by adjusting the concentration of the HCl solution , we controlled the doping concentration (Δp) of the ReSe2 between 3.64 × 1010 and 3.61 × 1011 cm−2. We subsequently confirmed that the VTH, µFE, ION, and photoresponsivity of the ReSe2 devices changed as the doping concentration increased in the Δp range: the ΔVTH increased from 3.06 to 15.41 V, the ratio of ION after/before the doping increased from 2.05 to 3.52, the ratio of µFE after/before the doping increased from 2.55 to 4.37, and the photoresponsivity at 520 nm increased from 79.99 to 314.4 A W−1. Using the selective HCl p‐doping process, the key performance indices of the ReSe2 photodetector were improved simultaneously, compared with the control ReSe2 photodetector. The photoresponsivity increased from 79.99 to 1.93 × 103 A W−1, and the rise and decay times were enhanced from 10.5 to 1.4 ms and from 291 to 3.1 ms, respectively, where VGS was 0 V and VDS was −5 V. This is because the p/p+ junction formed via selective HCl p‐doping of the ReSe2 channel region significantly affected the splitting of photocarriers and the injection of carriers from Pt to ReSe2. The proposed p‐doping technique and its fundamental analysis provide a scientific foundation for implementing high‐performance TMD‐based electronic and optoelectronic devices.We simultaneously enhanced the photo and temporal responses of the ReSe42HCl p‐Doping of ReSe: 0.1, 1, and 10 m HCl aqueous solutions were prepared by mixing saturated HCl aqueous solution (37% or 12 m) and deionized water. ReSe2 samples were then dipped into the prepared HCl solutions to dope ReSe2. After 5 min, the ReSe2 samples were rinsed with deionized water to remove residual of the HCl solution on the surface of ReSe2.2Raman and KPFM Analyses of Fully and Selectively HCl‐Doped ReSe: The HCl‐doped ReSe2 samples were analyzed through Raman spectroscopy, , and KPFM measurements. Raman spectroscopy with an excitation wavelength of 532 nm was used, where the laser beam diameter was ≈0.7–0.9 µm, the instrumental spectral resolution was less than 0.9 cm−1, and the integration time was 5 s. For the KPFM measurement, a platinum/iridium (Pt/Ir)‐coated Si tip was used and the tip was calibrated on a highly oriented pyrolytic graphite (HOPG) surface. The surface work function of the samples was obtained from the contact potential difference (CPD) between the tip and the HOPG work function (Φtip – ΦHOPG = VCPD), where the standard work function value of HOPG was used (4.6 eV). Further details on the KPFM analysis are provided in Figure S8 in the Supporting Information.Fabrication of the Control, Fully HCl‐Doped, and Selectively HCl‐Doped Devices: The ReSe2 crystals were commercially purchased and 35 nm thick ReSe2 layer was mechanically exfoliated to the 90 nm thick SiO2 on a heavily boron‐doped Si substrate by using adhesive tape . The remaining tape residue was then removed with acetone. For the control device, source/drain electrode regions were patterned by optical lithography (channel length and width: both 5 µm). Then, 10 nm platinum  and 40 nm palladium  layers were deposited in an e‐beam evaporation system. In case of selectively HCl‐doped ReSe2 devices, additional optical lithography process was conducted to block half of the ReSe2 region from HCl doping. After the selective HCl doping, the photoresist pattern was removed with acetone.2 Transistor DevicesCharacterization of the ReSe: The control and fully doped ReSe2 photodetector devices were electrically investigated through current–voltage measurements (ID–VG and ID–VD) by a Keysight B2912A precision source/measure unit. The threshold voltage (VTH), carrier concentration (Δp), and field‐effect mobility (µFE) were calculated from the measured data, where all drain currents (ID) were normalized by the channel width (5 µm). The carrier concentration and field‐effect mobility were respectively extracted using p = IDL/qWµVD and µFE = L/(WVDCOX) × (∂ID/∂VG), where L and W are the length and width of the channel, q is the electron charge, and COX is the capacitance of the gate oxide, respectively.2 Photodetector DevicesCharacterization of the ReSe: The control, fully doped, and selectively doped ReSe2 photodetector devices were investigated through the electrical measurement (ID–VG) under dark and illuminated conditions. The light sources were dot lasers with wavelengths of 520, 655, 785, 850, and 980 nm. The photoresponsivity (R) was calculated by the relationship, R = IPhoto/PLight, where IPhoto is the generated photocurrent and PLight is the total incident optical power. The photoresponse speed was analyzed under 520 nm laser illumination, which was turned on for 5 s and then off for another 5 s (1 cycle was 10 s).The authors declare no conflict of interest.SupplementaryClick here for additional data file.


Using a combination of 1s X-ray absorption (XAS) and 1s3p resonant inelastic X-ray scattering (RIXS), the valence, coordination and symmetry of cobalt ions were tracked in two cobalt-promoted molybdenum oxide precursors of the hydrodesulfurization catalyst system, namely Co–Mo/Al2O3 and Co–Ni–Mo/Al2O3. Extended X-ray absorption fine structure shows that the Co—O bonds were replaced with Co—S bonds as a function of reaction temperature. The cobalt K pre-edge intensity shows that the symmetry of cobalt was modified from Co3+Oh and Co2+Oh to a Co2+ ion where the inversion symmetry is broken, in agreement with a square-pyramidal site. The 1s3p RIXS data revealed the presence of an intermediate cobalt oxy-sulfide species. This species was not detected from XAS and was determined from the increased information obtained from the 1s3p RIXS data. The cobalt XAS and RIXS data show that nickel has a significant influence on the formation of the cobalt oxy-sulfide intermediate species prior to achieving the fully sulfided state at T > 400°C.The evolution in local structure and electronic properties of cobalt was investigated during  There are many factors that can affect the activity of a catalytic solid, e.g. the metal loadings, type of support and especially the synthesis conditions and treatment procedures employed et al.  oxide precursors. Among the many synthesis steps involved, the sulfurization process is very likely the step that determines the final structure and performance of the active Co–Ni–Mo/Al2O3 catalyst material. Following the analysis of Topsøe and Clausen, we start from the assumption that cobalt can occur in three forms in alumina-supported catalysts: (a) as cobalt ions on the edges of the MoS2 crystallites, (b) as Co9S8 particles and (c) as divalent cobalt ions at the surface or in the bulk of the alumina  catalysts has been one of the interesting research subjects in the field of heterogeneous catalysis because of the new environmental regulations to reduce sulfur emissions of transportation fuels  and 1s3p resonant inelastic X-ray scattering (RIXS). The 1s core levels of the 3d transition metal ions can be probed with hard X-rays (4–10 keV). In contrast to the main K-edge, which arises from the dipole allowed transition of 1s to 4p, the pre-edge involves mainly 1s3d quadrupole transitions. The K pre-edge energy position and intensity are sensitive to the metal oxidation state, the site symmetry, and the crystal-field splitting. However, because of their relatively low intensity and large core hole lifetime broadening, their analysis is often limited to the energy position and intensity of the pre-edge. In a 1s3p RIXS experiment, the resonant 1s3p X-ray emission is measured by tuning the energy of the incident energy to a 1s X-ray absorption edge. This effectively combines X-ray absorption and X-ray emission into a single experiment , whereas the spectral broadening of the 1s3p resonant XES depends on the final state 3p lifetime (Lp3)  and double  promoted HDS catalysts, using a combination of 1et al., 2004et al., 2010et al., 2013in situ conditions. In this work, RIXS is used to study the composition of the Co–Mo/Al2O3 and Co–Ni–Mo/Al2O3 HDS catalysts as model systems to reflect the abilities of the RIXS technique to study other heterogeneous catalysts under in situ conditions. The purpose of the present work is to unravel the nature of the cobalt species in the precursor oxide of γ-Al2O3-supported Co–Mo and Co–Ni–Mo catalysts and to follow the changes in the local structure and electronic properties, during the in situ sulfurization with RIXS and XAS.Hard X-rays are able to penetrate solids and probe the bulk phase properties of the samples  with a solution of metal salts. Aqueous solutions of ammonium heptamolybdate hexahydrate , cobalt nitrate hexahydrate  for Co–Mo/Al2O3 and additionally nickel nitrate hexahydrate  for Co–Ni–Mo/Al2O3 were co-impregnated to obtain weight loadings of 14 wt% for molybdenum and 5 wt% for both cobalt and nickel. Subsequently, both samples were dried at 60°C for 16 h then at 120°C for 1 h. To convert the impregnated metal precursors to their respective metal oxides, the samples were calcined at 450°C for 16 h. All drying and calcination steps were performed under a (75% N2)/(25% O2) flow (1 ml min−1). We note that the atomic Co/Mo ratio of 0.58 is higher than used in the industrial HDS catalysts. We used the increased amount of cobalt and nickel to increase the signal-to-noise of the X-ray spectroscopy experiments.The Co–Mo/Alin situ sulfurization experiments both oxide precursor samples were simultaneously sulfurized by loading the samples on the same sample holder. Pellets of Co–Mo/Al2O3 and Co–Ni–Mo/Al2O3 samples were made from the respective powders and loaded on the same sample holder as is shown in Fig. 1−1, while flowing with a 10% H2S/H2 mixture (1 ml min−1). The samples were kept at 450°C for 5 h.During the in situ sulfurization under dynamic flow of 10% H2S/H2 gas mixture and by slowly heating the samples to 450°C.The reactor was assembled and sealed and the incident window was covered with Kapton foil to decrease the photon flux and prevent radiation damage and leakage of air into the reactor. The reactor includes connections for both gas inlet and gas outlet. The inlet line is connected with a mass-flow controller that regulates the type and amount of gas that passes through the reactor. On the outlet side, there are bubbler airlocks that can be used to trap the toxic gas mixture. First, the oxide precursors were measured under ambient conditions, followed by 2.2.et al. 2013K-edge. After the monochromator, the beam is delivered to the Rh-coated focusing mirror and focused both horizontally and vertically to ∼140 µm × 400 µm (V × H) at the sample position. XAS spectra were measured simultaneously in total fluorescence yield (TFY) mode using a photodiode. The sample, crystal analyzer and photon detector were arranged in a vertical Rowland geometry. The Co Kβ (1s3p) RIXS spectra were recorded at a scattering angle of 90° in the horizontal plane using seven analyzer crystals  of the beamline and the spectrometer is ∼0.2 eV. The intensity was normalized to the incident flux. The RIXS data are shown as a contour map in a plane of incident and transferred photon energies, where the vertical axis represents the energy difference between the incident and emitted energies (energy-loss). The variations in colour on the plot correspond to the different scattering intensities.The spectra were recorded at beamline 6-2 of the Stanford Synchrotron Radiation Lightsource (SSRL)  was performed under 0.1 µl of thiophene saturated in 10 ml min−1 H2, at a temperature of 320°C. The sample was then stabilized using 1 µl thiophene saturated H2 with a flow rate of 10 ml min−1 at 320°C. Following a stabilization step, the catalytic activity of the samples was measured under similar conditions. The gaseous products were analyzed online on a mass spectrometer through a sampling and a flame ionization detector. Fig. 32O3 and Co–Mo–S/Al2O3 during three subsequent HDS steps. Since variation in molybdenum-loading influences the HDS activity, the conversion is normalized for the molybdenum loading of the catalysts. An initial conversion of 27% was observed for the double promoted catalyst and 60% for the single promoted catalyst material. The thiophene conversion data were normalized to the Mo loading.Before presenting the results of the 2O3, an increase in the formation of bulk cobalt- and nickel sulfide phases is expected, resulting in a reduced catalytic activity. The lower conversions observed for Co–Ni–Mo–S/Al2O3 can be explained by a decrease in the number of promoter atoms on the edges of MoS2 slabs. In addition, as shown in Fig. 3Because of the higher ratio of promoters to molybdenum in Co–Ni–Mo–S/Al3.2.supporting information. Fig. 4K-edge X-ray absorption near-edge (XANES) region as a function of temperature. The cobalt K-edge spectrum of freshly calcined Co–Mo/Al2O3 is characterized by an intense main edge (1s → 4p) and a weak pre-edge feature with a main peak at 7709 eV followed by a weak shoulder at 7711.4 eV. This finding is similar to the previously reported Co3O4 reference . The Co–Mo/Al2O3 sample shows the complete sulfidation at 190°C, whereas the Co–Ni–Mo/Al2O3 sample is only fully sulfided at 400°C. Furthermore, these results are confirmed by the increase in the integrated pre-edge intensities and the analysis of the extended X-ray absorption fine structure (EXAFS) region during the sulfurization process, which was sensitive to mainly the presence of either Co—O or Co—S bonds. The EXAFS analysis illustrates the considerable impact that the temperature has on the sulfurization process and on the formation of Co—S bonds under the flow of the 10% H2S/H2 gas mixture . At room temperature, approximately 45% and 50% of the total bonds are converted to Co—S bonds for Co–Mo/Al2O3 and Co–Ni–Mo/Al2O3 samples, respectively. This increase is attributed to gradual ligand substitution and conversion of cobalt ions at Oh sites in the case of the oxide precursor to the (Cv4) square-pyramidal cobalt in symmetry for the fully sulfided sample.In the performed composition analysis (listed in Tables S2 and S3), the Co–Mo/Al3.3.s3p RIXS planes were recorded during the in situ sulfurization of Co–Mo/Al2O3 and Co–Ni–Mo/Al2O3. Fig. 6s3p RIXS spectra of Co–Mo/Al2O3. The equivalent 1s3p RIXS spectra of Co–Ni–Mo/Al2O3 are shown in Section S3. The figure shows several changes during the in situ sulfurization reaction, including (a) the gradual conversion of two distinct pre-edge features to a single emission feature and (b) a shift of the main edge to lower absorption energies.A series of cobalt 12O3 catalyst are shown in Figs. S4 and S5. The oxide precursor has distinctive resonance peaks at 7650.7 eV (peak 1) and 7647.45 eV (peak 2) characteristic of divalent cobalt species  by holistically fitting all spectra. We performed 100 fits in each stage of the in situ reaction using the methodology implemented in Blueprint XAS to explore uncertainty of all fit parameters  and the sample at 400°C for each temperature in addition to a small offset value to account for the counts on the tails of other emission processes. Initially, to fit the RIXS data, the combination of only two components was used to analyse the spectra. However, this proved to be impossible as additional peaks were observed in the experimental spectra during the 3.4.2O3 sample the RIXS data analysis revealed the presence of a single intermediate species (in blue) for the lower temperatures (T < 150°C). Because of its spectral shape, we can assume a species consisting mostly of Co—S bonds. However, the fitting of the Co–Ni–Mo/Al2O3 RIXS data was only possible by the inclusion of two intermediate species (blue and purple), which remain present during the in situ reaction for T < 400°C. The shape of one of the species has a predominantly oxide character [blue in (a)], while the second has a rather sulfide character [purple in (b)]. This assumption is based upon their comparison with the spectra of the pure oxide and the pure sulfide with regard to their individual shape as well as the position and intensity of the spectral features. In other words, in Co–Mo/Al2O3 the intermediate is most likely to be the Co—S5O and/or Co—S4O2 oxy-sulfide, which can be distinguished from Co—S6 in some intermediate stages. In Co–Ni–Mo/Al2O3 we see both the Co—S5O and/or Co—S4O2 oxy-sulfide (blue) and also the Co—S1O5 and Co—S2O4 oxy-sulfides (purple).The obtained phase concentrations are shown in Fig. 82O3 sample is more sensitive toward sulfidation at room temperature as 60% (versus 30% for Co-Mo/Al2O3) of the oxide species is already converted to either the sulfide or intermediate species under the dynamic flow of the gas mixture at 25°C. The details of the analysis and all numbers are given in Tables S4 and S5. We note that at 400°C oxidic cobalt is no longer detected, implying the absence of divalent cobalt ions in the alumina. In addition, a fraction of the sulfide phase could exist as Co9S8 particles.The Co–Ni–Mo/Al4.2O3 and Co–Ni–Mo/Al2O3 samples, an increase in the pre-edge intensity was observed. The increase is caused by the conversion of cobalt species to a species without inversion symmetry, for example in square-pyramidal symmetry. Furthermore, because of the sulfurization process the cobalt was reduced and the Co K main edge was also shifted to lower absorption energies. The EXAFS data analysis showed the gradual ligand exchange during the process of sulfurization. The relative concentration percentages of Co—O and Co—S bonds were determined by fitting the EXAFS data as the linear combination of components at the two extremes. The analysis showed a higher tendency towards sulfurization of cobalt in the case of Co–Mo/Al2O3. In addition, the gradual change in the composition was obtained for the subsequent steps of the in situ reaction.During the gradual sulfurization of cobalt in the Co–Mo/Als3p RIXS study has shown a gradual transition of the cobalt oxide in the active phase to the fully sulfided species. One of the findings is that the Co–Mo/Al2O3 catalyst has a higher tendency for the sulfurization reaction and achieves a >90% conversion to the sulfide phase at 190°C, compared with 400°C for the Co–Ni–Mo/Al2O3 catalyst. By analyzing the fits obtained from the RIXS data, the existence of a single intermediate species for T ≤ 150°C with a sulfide-like character was detected for the Co–Mo/Al2O3 catalyst. In the case of the Co–Ni–Mo/Al2O3 catalyst two intermediates were identified that eventually converted to the fully sulfided phase at 400°C. The RIXS data confirmed the previously obtained results by XANES and EXAFS analysis of the two samples. A summary of the proposed cobalt species involved during the in situ sulfurization of the Co–Mo/Al2O3 and Co–Ni–Mo/Al2O3 HDS catalysts is shown in Fig. 9et al.  the catalytic activity testing, (2) the XAS data analysis method, (3) the numerical XANES fit results and (4) the 110.1107/S1600577519002546/hf5380sup1.pdfSupporting information . DOI:


This combined kinetic and structural study provides a foundation to develop next‐generation hairpin designs where the DNA‐binding profile of polyamides is reconciled with their physicochemical properties.A combined structural and quantitative biophysical profile of the DNA binding affinity, kinetics and sequence‐selectivity of hairpin polyamide analogues is described. DNA duplexes containing either target polyamide binding sites or mismatch sequences are immobilized on a microelectrode surface. Quantitation of the DNA binding profile of polyamides containing N‐terminal 1‐alkylimidazole (Im) units exhibit picomolar binding affinities for their target sequences, whereas 5‐alkylthiazole (Nt) units are an order of magnitude lower (low nanomolar). Comparative NMR structural analyses of the polyamide series shows that the steric bulk distal to the DNA‐binding face of the hairpin  N‐methyl pyrrole (Py) and N‐methyl imidazole (Im) heterocyclic amino acids defines the selectivity of dsDNA binding ranging from 7 up to 24 base‐pairs in length  are cell‐permeable transcriptional modulators which function by inhibiting RNA polymerase‐mediated elongation and/or transcription factor binding to its target double‐stranded DNA (dsDNA) consensus sequence.PA2–3) direct a hydrogen‐bond‐acceptor (N3) atom towards the floor of the minor groove and forms a hydrogen bond with the exocyclic hydrogen bond donor amine (N2) of G. A key structural difference with the incorporation of an Nt‐unit in the N‐terminal position of a hairpin PA is the endocyclic sulfur atom which changes both the geometry and hydrophobicity (logD) of this heterocycle , a more pronounced compression of the major groove is observed relative to the archetypical hairpin PA1⋅dsDNA complex.We have recently expanded the heterocyclic repertoire of current Py‐Im hairpin PA designs to include N‐terminal thiazole‐4‐carboxylic acid units (Nt).e Figure .7 Furthe⋅dsDNA interactions where the N‐terminal heterocycle is systematically altered (PA1–4). PAs containing N‐terminal Im units  exhibit enhanced selectivity for their target sequences relative to cognate Nt units . Whilst increasing the steric bulk of the iPr‐Im unit (PA4) does not impact DNA binding affinity for its target sequence, NMR structural analysis reveals the larger iPr‐Im unit does induce more pronounced structural perturbation of the target dsDNA duplex relative to PA1, which contains an N‐terminal Me‐Im unit.In this manuscript, we report a label‐free biophysical assay to profile the affinity, sequence‐selectivity and binding kinetics of PAPA1) was chosen as our exemplar scaffold to explore the dsDNA binding profile as a function of four different N‐terminal heterocycles.PA1 has an established high affinity binding profile for the general sequence 5′‐WWGWWCW (where W=A/T), for which we used 5′‐GTACTAT as the target sequence in an immobilized DNA duplex (ODN1).PA1–4 were prepared using Boc‐based solid phase synthesis on a β‐Ala PAM resin via amide coupling of the corresponding heterocyclic carboxylic acid (Scheme S1).The heterocyclic core of a known hairpin PA sequence kon and koff) and the equilibrium dissociation constant (KD).Tm) of PA⋅ODN complexes as a function of a temperature gradient.A schematic of the experimental setup is shown in Figure PA1–4 to ODN1 show all four PAs exhibit high‐affinity binding (Table PA1 and PA4) exhibit KD values in the picomolar range, the Nt‐containing PAs (PA2–3) exhibited a binding affinity that is approximately an order of magnitude lower . Rate maps of PA1–4 targeting ODN1 provided deeper insight into the origin of the differences in the KD values of our PA set  for each PA was similar, the rate of association (kon) of PA2–3 was approximately an order of magnitude slower relative to PA1 and PA4.Kinetic analyses of the binding profile of  PAs PA2– exhibitePA1–4 was explored using duplexes where the target binding sequence in ODN1 was replaced with mismatched sequences (ODN2–3). Analyses of the binding kinetics show that Im‐containing PAs (PA1 and PA4) are more G‐selective relative to Nt‐containing PAs  of PA4 for all sequences ODN1–3 were similar, the dissociation rates (koff) were significantly faster for mismatched sequences (ODN2–3). A less pronounced kon/koff trend was observed for PA1 binding to ODN2, while no interaction was measured with ODN3.The sequence selectivity profile of PA2  and the mismatch (ODN2) sequence. Out of the PA series, PA3 displayed the most unique binding profile  for free ODN1–3 and in complex with 20 nmPA1–4. The UV/Vis melting profiles of the PA⋅dsDNA complexes confirm a similar trend in dsDNA sequence selectivity  observed in the fluorescence experiments . Of particular note was the melting stabilization of PA4, which displayed excellent G‐selectivity relative to PA1–3. Consistent with our kinetics profiling  is superior to Nt‐containing analogues (i.e. PA2–3). Furthermore, whilst enhancing steric bulk on the 5‐position of the Nt‐series enhanced G‐selectivity, this had little effect on the Im‐series .This experimental setup was also used to determine duplex stabilization of PAiPr‐Im unit incorporated in PA4 when in complex with its target dsDNA sequence, NMR studies were undertaken using the self‐complementary dodecamer sequence d(CGGTACAATTCG)2 (ODN4). Titration experiments of PA4 into a solution of ODN4 confirmed the formation of a 1:1 PA4⋅ODN4 complex. 2D NOESY studies at 4 different mixing times identified a suite of strong NOE cross‐correlations from H4 of the iPr‐Im building block to G5H1 and the G5N2 exocyclic amine, which implies that the iPr‐Im N3 is directed towards the floor of the minor groove  relative to PA1⋅ODN4. A similar trend in enhanced major groove compression was also observed with PA4⋅ODN4 relative to PA1⋅ODN4  relative to PA3⋅ODN4 but it is not as extensive as that observed for the PA1⋅ODN4 complex (2.01 Å).The origins of these differences become apparent when comparing the extent of minor groove penetration of the three complexes Figure . NMR‐res2 Figure  a.13 In 2 Figure  b.13 TheThis combined kinetic and structural study has shown that the type of N‐terminal heterocycle and its substituents influences the dsDNA binding profile and the overall structure of the duplex. We discuss here several conclusions that emerged from our results.PA1/PA4) showed a higher binding affinity relative to the Nt‐containing PA2–3 via an increase in the rate of association. Although there has not been a study dedicated to evaluating the influence of the hairpin PA N‐terminus, a previous SPR‐based study by Sugiyama et al. has shown that the number of Me‐Im and their positioning in a hairpin PA scaffold can have a disproportionate impact on the ak and DK relative to only small changes in the kd.β‐alanine units influences both ak and dk parameters.Firstly, all four PAs exhibit high affinity (low nanomolar‐picomolar) for its target dsDNA sequence. However, the two N‐terminal Im‐containing PAs , an improvement in G‐selectivity relative to the iPr‐Nt unit  is likely due to a greater level of minor groove penetration  should be addressed to the authors.SupplementaryClick here for additional data file.


ALMT1; exuded malate is suspected to interact with extracellular Fe to inhibit root growth. In addition, ALS3 – an ABC‐like transporter identified for its role in tolerance to toxic Al – represses nuclear accumulation of STOP1 and the expression of ALMT1. Until now it was unclear whether Pi deficiency itself or Fe activates the accumulation of STOP1 in the nucleus. Here, by using different growth media to dissociate the effects of Fe from Pi deficiency itself, we demonstrate that Fe is sufficient to trigger the accumulation of STOP1 in the nucleus, which, in turn, activates the expression of ALMT1. We also show that a low pH is necessary to stimulate the Fe‐dependent accumulation of nuclear STOP1. Furthermore, pharmacological experiments indicate that Fe inhibits proteasomal degradation of STOP1. We also show that Al acts like Fe for nuclear accumulation of STOP1 and ALMT1 expression, and that the overaccumulation of STOP1 in the nucleus of the als3 mutant grown in low‐Pi conditions could be abolished by Fe deficiency. Altogether, our results indicate that, under low‐Pi conditions, Fe2/3+ and Al3+ act similarly to increase the stability of STOP1 and its accumulation in the nucleus where it activates the expression of ALMT1.Low‐phosphate (Pi) conditions are known to repress primary root growth of Arabidopsis at low pH and in an Fe‐dependent manner. This growth arrest requires accumulation of the transcription factor STOP1 in the nucleus, where it activates the transcription of the malate transporter gene  Low‐phosphate, low‐pH and excess aluminum are three major stresses inhibiting root growth. The Arabidopsis transcription factor STOP1 has a major role in root growth under these different stresses, but how STOP1 is regulated is not known. Here we show that Fe –instead of low‐phosphate– and Al promote the accumulation of STOP1 in root nuclei, in a low‐pH‐dependent manner. This work unveils an important regulatory step in the response to Al and Fe stresses. In many plant species, phosphate (Pi) deficiency (–Pi) alters root growth and architecture, promoting top‐soil foraging: the growth of the primary root is reduced whereas lateral root emergence and growth is stimulated toward a more horizontal direction. Combined, these responses result in root systems that explore relatively better the upper soil horizons where Pi is more concentrated  and PHR1‐like 1 (PHL1), the –Pi local response is not , a short primary root results from a combination of rapid as well as long‐term cellular responses at the root tip. A few hours, if not minutes, after the root tip encounters a Pi‐depleted zone, cell elongation decreases. This is correlated with the deposition of Fe and the accumulation of reactive oxygen species (ROS) in the cell wall of the stem cell niche (SCN) and the elongation zone (EZ), and with a peroxidase‐dependent stiffening of the cell wall of the EZ.et al., et al., et al., In the long term (hours to days), callose accumulates in the cell wall of the root apical meristem (RAM) and EZ, this occludes plasmodesmata and restricts cell‐to‐cell trafficking. Progressively, root cell proliferation ceases until exhaustion of the RAM by the on‐going cell differentiation (Muller LOW PHOSPHATE ROOT 1 (LPR1) encodes a cell wall multi‐copper oxidase with ferroxidase activity  and SCARECROW (SCR). This regulation operates via increased expression of CLAVATA 4 (CLV4), a secreted peptide detected by the plasma membrane receptors CLV2 and CLV2/PEP1 RECEPTOR 2 (PEPR2)  is unrepressed in –Pi and expression of LPR1 is reduced  gene was identified in the local response  , encoding a plasma membrane transporter exuding malate  interacts with STAR1 (SENSITIVE TO ALUMINUM RHIZOTOXICITY 1) to form a putative ATP‐binding cassette (ABC) transporter complex located in the tonoplast   construct to investigate the role of Fe because this visual reporter sensitively allows to test the activity of STOP1 signaling in the root tip  during this pre‐culture, the growth medium was buffered at pH 6.7. Indeed, at a pH around neutrality we observed no or very little GUS staining in the root tip of pALMT1::GUS seedlings , the expression of ALMT1 was still induced. This result confirms that ALMT1 expression is higher in –Pi than in +Pi conditions. However, increasing the Fe content in a medium containing 250 μm Pi increases the expression of ALMT1   Table .ALMT1 expression, we reduced the availability of Fe in the growth medium using a siderophore. Supplementing the medium with 100 μm deferoxamine (DFO), a potent Fe chelator, strongly diminished the expression of ALMT1 , a classical marker of –Pi stress. SPX1 encodes a protein regulating the activity of the transcription factor PHR1 – a master regulator of –Pi stress which is frequently used as a marker of the –Pi transcriptional response. To monitor its expression, we used WT seedlings containing the pSPX1::GUS marker  gene as an additional marker whose expression is highly stimulated by Pi deficiency  gene, a well‐known indicator of Fe deficiency . We then tested the effect of MG132, an inhibitor of the 26S proteasome, on the accumulation of GFP‐STOP1 in seedlings grown in a moderate amount of Fe  is required for Al to promote the nuclear accumulation of GFP‐STOP1  , STOP1 is poorly accumulated in the nucleus. Then, if STOP1 does not accumulate in nucleus under these growth conditions, how does it participate in resistance against low pH? One hypothesis is that at pH 4.3 a very low amount of Fe is sufficient to stimulate the accumulation of STOP1 for triggering the expression of genes for H+ tolerance. Alternatively, STOP1 might have already started promoting the expression of genes involved in H+ tolerance before (i.e. during embryogenesis) seedlings encounter acidic conditions. A transcriptomic study of seedlings grown in an acidic environment showed that the stop1 mutant is defective for the expression of several genes, including genes involved in cell wall composition or modeling  or a Fe‐ (or Al3+)‐containing molecule. This hypothesis would fit with the role of ALS3–STAR1 in Al3+ tolerance. However, we cannot exclude that this metabolite does not contain Fe (or Al3+).We showed here that overaccumulation of STOP1 in the nucleus of m Figure . Since oSTOP1, ALS3 and STAR1 genes are all expressed in the root tip  activates the STOP1 branch.A two‐branched regulatory pathway modulates root growth arrest: the STOP1/ALMT1/ALS3/STAR1 branch and the LPR1/PDR2 branch  regulates STOP1.For Al, the model only partially applies because the exuded malate prevents the inhibitory effect of toxic Alet al., et al., et al., et al., et al., et al., Few Fe‐sensing proteins have been identified in plants , pALMT1::GUS#2, pSTOP1::GFP‐STOP1#B10 and als3; pSTOP1::GFP‐STOP1#B10 lines were described previously  and agarose (7 g L−1) for plates were from Sigma‐Aldrich  and Lonza , respectively. The media were buffered at pHs ranging from 5.5 to 6.7 with 3.4 mm 2‐(N‐morpholino)ethanesulfonic acid and at pH 7.1 with 3 mM 1,4‐piperazinediethanesulfonic acid disodium salt. The different plates were prepared by mixing the melted autoclaved 1× agar or agarose medium (composition as above) with the buffer. Solutions of AlCl3 and FeCl2 were filtrated before adding to the plates.Seeds were surface‐sterilized for 5 min in a solution containing 70% ethanol and 0.05% sodium dodecyl sulfate, and washed twice with 96% ethanol. The nutrient solution was the same as used previously , itself lying on the agar plate. After 3 days of growth on the agar plate, the nylon meshes carrying the seedlings were transferred to the new plates containing, or not, different concentrations of Al or Fe.For the experiments with MG132, nylon meshes carrying the 3‐day‐old seedlings were transferred on a 10‐μl droplet (liquid extracted from Sigma‐Aldrich agar A1296 #BCBL6182V from which the seedlings were pre‐grown) containing, or not, different concentrations of MG132. In all experiments, the pH of the growth medium for germination and for the subsequent transfer was identical.For the qRT‐PCR experiment, seeds were sown side by side in a line, on a 5‐cm strip of nylon meshes. Three to 3.5 days after sowing the whole roots of seedlings were harvested and mRNA extracted.m of DFO was added to the 1× melted autoclaved growth medium just before pouring the plates. For the washed DFO‐treated agar, we prepared the agar as follows: 4 g of Sigma‐Aldrich agar (A1296 #BCBL6182V) was added to 100 ml of MilliQ water containing 600 μm of DFO and stirred for 15 h. The agar was then washed by centrifugation (520 g) through a nylon mesh (mesh size 10 μm); this washing step with MilliQ water was carried out twice. This washed DFO agar was then used to prepare the growth plates.We used two methods to reduce bioavailable Fe in the growth medium. For the DFO agar, 100 μ3  by incubation overnight in an oven at 80°C. The solutions were diluted to a final volume of 5 ml by the addition of distilled water. The Al, Fe and P were quantified using ICP‐AES . The concentrations of metals were determined using standard curves obtained from solutions made with ICP‐grade elements.Dried samples of agars and agarose were mineralized by the addition of 300 μl of 70% HNOhttps://www.zymoresearch.com/) and treated with the RNase‐free DNase Set (Zymo Research) according to the manufacturer's instructions. Reverse transcription was performed on 500 ng of total RNA using qScript™ cDNA SuperMix . Quantitative PCR was performed on a 480 LightCycler thermocycler  using the manufacturer's instructions with Light Cycler 480 SYBR Green I Master (Roche) and the primers listed in Table At5g62690) as a reference gene for normalization and the quantification of gene expression was as according to published method  using a ×20 dry objective (Plan Apo NA 0.80). The GFP was excited with an Ar ion laser (488 nm). Emitted light was collected from 493 to 538 nm for GFP, using the MBS 488 filter. All nuclei were imaged using the same conditions of gain, offset and resolution and with a pinhole setting of 1 a.u. The quantification of GFP fluorescence was carried out as follows: a Z‐stack of nine images (separated by a distance of 1.8 μm) imaged representative nuclei on the surface of the root. Images were acquired in 12 bits using Zen black software , then converted to a maximum projection image. The average nuclear fluorescence intensities were quantified using Zen blue software , by drawing identical regions of interest. A 120 μm × 60 μm rectangle was defined at 200 μm from a root tip (roughly in the transition zone), in which a region of interest (6 μm × 6 μm) inside each nucleus was defined for the measurement of fluorescence.Images were collected on a Zeiss LSM780 confocal microscope .Table S2. Primer sequences.Click here for additional data file. Click here for additional data file.


Focusing X‐rays to single nanometer dimensions is impeded by the lack of high‐quality, high‐resolution optics. Challenges in fabricating high aspect ratio 3D nanostructures limit the quality and the resolution. Multilayer zone plates target this challenge by offering virtually unlimited and freely selectable aspect ratios. Here, a full‐ceramic zone plate is fabricated via atomic layer deposition of multilayers over optical quality glass fibers and subsequent focused ion beam slicing. The quality of the multilayers is confirmed up to an aspect ratio of 500 with zones as thin as 25 nm. Focusing performance of the fabricated zone plate is tested toward the high‐energy limit of a soft X‐ray scanning transmission microscope, achieving a 15 nm half‐pitch cut‐off resolution. Sources of adverse influences are identified, and effective routes for improving the zone plate performance are elaborated, paving a clear path toward using multilayer zone plates in high‐energy X‐ray microscopy. Finally, a new fabrication concept is introduced for making zone plates with precisely tilted zones, targeting even higher resolutions. MLLs arose from the difficulty of fabricating FZPs with tilted zones, which are composed of a set of planar linear‐zone plates in a crossed geometry. MLLs suffer from even more stringent alignment requirements due to the chromatic aberration. Very recently, 2D sub‐10 nm resolution has been reported, reinforcing the potential of multilayer optics.One of the most widely used and successful high‐resolution X‐ray focusing optics is the Fresnel zone plate (FZP). It consists of alternating opaque and transparent coaxial annuli, which form the zones. The spatial Rayleigh resolution of an FZP is defined by, ΔδAn attractive approach toward realizing high‐resolution hard X‐ray FZPs is to coat a cylindrical substrate of a well‐defined, smooth surface with alternating layers of proper material combinations,Since the first trials in the 1980s,A recent method for producing ML‐FZPs is based on atomic layer deposition (ALD) of multilayers over optical quality glass fibers.Figure2O3–HfO2 ML‐FZP of extremely high quality. We investigated its properties by a variety of characterization techniques and a 15 nm half‐pitch cut‐off resolution was recorded via direct imaging experiments in a scanning transmission soft X‐ray microscopy (STXM) set‐up. Encouraged by this success, we introduce a new fabrication concept for making ML‐FZPs with precisely tilted zones. We discuss the implications of tilted FZPs, from coupled wave theory simulations (CWT) and argue why they are crucial for achieving higher resolution FZPs. Finally, the first direct imaging and diffraction efficiency results obtained by our proposed concept are demonstrated.In this paper, we followed a fully conformal, 3D bottom‐up growth approach based on ALD and subsequent focused ion beam (FIB) slicing Figure1. Thus22.12O3–HfO2 material couple offers high diffraction efficiencies for both soft and hard X‐rays and therefore were selected as the materials of the ML‐FZP. Another advantage of Al2O3 and HfO2 is their high melting temperatures of 2072 and 2758 °C respectively, providing a high chemical stability.Theoretical diffraction efficiencies of ML‐FZPs consisting of material pairs available to ALD have been calculated to identify best material couples, and were previously published.2O3–HfO2 multilayers were deposited via ALD at 290 °C on several optical quality glass fiber cores  to space (Al2O3) ratio was used instead of the standard 1:1. This allowed for a faster deposition due to shorter Al2O3 deposition cycle .The Al Figure  a,b, usin+ FIB as demonstrated schematically in Figure  + FIB, which is optimized for the soft X‐ray range. An additional Pt beam‐stop layer of diameter d = 25 µm was deposited in the center of the FZP, also by FIBID as illustrated in Figure  d = 39.4 µm, outermost zone width of Δr = 25 nm, inactive central obstruction of dco = 31.4 µm with 4 µm of zone thickness and an aspect ratio of 28. The scanning electron microscopy (SEM) images of the ML‐FZP, prior to FIBID of Pt beam‐stop are shown in Figure  The deposited glass fibers Figure  c were sl2.2FigureTo investigate the zone quality in the optical axis, a rectangular lamella was lifted out in the FIB from the deposited glass–fiber as depicted in 4,5 edge intensities, the interfaces of Al2O3 and HfO2 were further confirmed to be chemically abrupt. This is attributed to the chemically stable nature of the amorphous ceramic films deposited via ALD, lacking fast diffusion paths such as grain boundaries. It has been shown that amorphous films, if dense enough, are better diffusion barriers due to lack of grain boundaries.The energy‐dispersive X‐ray spectroscopy (EDX) elemental mapping of Figure  2O3 and HfO2 by X‐ray photoelectron spectroscopy (XPS) , EDX  analysis and determined to be ρ = 3.0 g cm−3 and ρ = 8.9 g cm−3 for Al2O3 and HfO2 thin films, respectively .The chemical composition of the thin films were estimated to be 38 at% Al to 62 at% O for alumina films and 34 at% Hf to 66 at% O for hafnia films by wavelength‐dispersive X‐ray spectroscopy (WDX). Chemical compositions were further confirmed to be Al2.3Inaccuracies in the zone pattern may reduce the resolution and diffraction efficiency of the FZP and cause aberrations. The effect of eccentricity, nonconcentric zones, radial displacement of the zones and zone roughness have been discussed elsewhere.r) by nature of the fabrication method. The eccentricity (ΔR/R) was estimated to be 0.0032 and within the tolerance valuesR/R < 0.35N−1) according to the measurements from the SEM image of Figure  The ALD ML‐FZPs with glass fiber cores are expected to have concentric zones (concentricity: γ < 1.2Δr). However, detailed characterization of the glass fiber cores show that the diameter of different glass fibers vary between 30 and 31.5 µm. The zone plate of interest of this paper has a measured core diameter of 31.4 µm. However the multilayer design was made for a FZP core of 30 µm. This causes a systematic radial displacement of the zones by 700 nm. If the zone thicknesses are accurate, the systematic shift of the zones arising from nonexact diameter of the inactive core, is expected to slightly modify the distribution of the field intensity of the focusing ring.However, if the deposition thicknesses are not strictly controlled, the stochastic nature of the bottom‐up fabrication approach2.4The diffraction efficiency and resolving capability of the ML‐FZP was tested in the soft X‐ray range at a state‐of‐the‐art STXM, MAXYMUS (UE‐46 PGM‐2) beamline located at the synchrotron facility BESSY II.FigurePrior to imaging experiments, the ML‐FZP was aligned in pitch and yaw using an in‐house constructed tilt stage until a full 1st order diffraction ring was achieved. It is known that the presence of a central obstruction causes the intensity in the focal spot to shift to side lobes.Table−3 from weighted mean mass density of the elements. The CWT calculations were done for the outermost period locally. Interdiffusion and roughness were neglected in accordance with the HRTEM, TEM‐EDX, and EELS results. For the calculation, the following parameters were considered: Outermost zone width Δr = 25 nm, densities of ρ = 8.9 g cm−3 and ρ = 3.0 g cm−3  and line to space ratio of 1:2 for HfO2 and Al2O3 thin films, respectively. To be able to compare measured and expected diffraction efficiencies a constant optical thickness of 700 nm was considered for the calculation of expected diffraction efficiencies in the soft X‐ray range. The expected performance of the optic in hard X‐rays is included in the Table  The diffraction efficiency measurements were conducted via scanning a pinhole over the FZP Figure  b at vari32O3–HfO2 FZP with an outermost zone width of Δr = 25 nm, a diameter of d = 39.4 µm and a deposition thickness of 4 µm was successfully fabricated. ALD multilayers constituting the zones exhibited chemically abrupt and structurally sharp interfaces on the molecular level. The ML‐FZP focusing performance was studied in detail in the soft X‐ray range at a state of the art STXM (UE46‐PGM2) located at the synchrotron radiation facility BESSY II. The own built soft X‐ray microscope was chosen as the experimental set‐up due to its versatility and stability allowing for highly precise characterization of the focusing performance. The imaging tests revealed a half‐pitch cut‐off resolution of 15 nm for the first time for a ML‐FZP. This is a marked increase over previous related research.A ML AlFigureThe diffraction efficiency maps as a function of energy and zone plate thickness were calculated for up to 30 keV using the thin grating approximationOne concern in the X‐ray optics community is that sputter‐sliced FZPs in general, tend to have zones with high roughness and low circularity, which has been shown to be detrimental to the imaging capabilities.2O3 (m.p. = 2072 °C) and HfO2 (m.p. = 2758 °C) layers are desirable properties for withstanding intense X‐ray pulses of XFELs and emerging diffraction limited synchrotrons. Furthermore, the relatively low X‐ray absorption of Al2O3 and HfO2 would result in less heating during intense X‐ray pulses, improving their life‐time.The high melting temperatures and radiation resistance of AlThe electron microscopy analyses confirm the zones to be fully amorphous. The advantage of amorphous layers is two‐fold. First, amorphous layers eliminate the possibility of undesired scattering from the crystallites, particularly important for hard X‐ray focusing. Second, the lack of grain boundaries ensure that there are no fast diffusion paths available which could lead to interdiffusion between the layers and decrease the performance.−1,A current disadvantage of the method that the diameters being small due to the slow deposition rates, can be overcome by using spatial ALD, that allows for deposition speeds up to 3.6 µm h3.1r . Using gold as the central obstruction makes the need of depositing a beamstop by means of FIBID obsolete, hence avoids the complications discussed in the first part of this work.The theoretical local diffraction efficiency of the outermost zone period of the suggested alumina/silica ML‐FZP is shown in Figure  FigureThe performance of a tilted ML‐FZP was tested also at the MAXYMUS UE‐46 PGM‐2) beamline. The measured diffraction efficiency of the tilted ML‐FZP achieved 11% of the theoretically expected value in an X‐ray energy range between 800 and 1450 eV Figure6a, and PGM‐2 be4Preparation of ML‐FZPs with extraordinary zone quality via ALD of amorphous–ceramic multilayers over optical glass fibers and subsequent FIB slicing was demonstrated. Consequently, the ML‐FZPs achieved up to 80% of their theoretical efficiency and enabled 15 nm half‐pitch cut‐off resolution in direct imaging experiments, which is remarkably close to the theoretical predictions. We also introduced a new fabrication strategy, which allows the fabrication of ML‐FZPs with precisely tilted zones. To the best of our knowledge, this is the first demonstration of a manufacturing route that delivers FZPs with precise tilt‐angles. Furthermore, we demonstrated that the proposed fabrication concept enables the tilting angle to be tailored arbitrarily, and enables fabrication of FZPs with different tilt angles simultaneously. We have shown the first imaging and efficiency analysis using the tilted ML‐FZPs and resolved the smallest 30 nm features of the Siemens Star test sample. Based on the presented experimental results and calculations, it is expected that single nanometer resolutions with high‐diffraction‐efficiencies will soon be available in the soft and hard X‐ray regime with the atomic layer deposited, ceramic ML‐FZPs.5Atomic Layer Deposition: Optical quality glass fibers from Schott AG (Germany) with the product code A2/30 µm were used as the core. The Al2O3–HfO2 thin films were deposited with a SENTECH SI ALD LL instrument (Germany) at 290 °C. For the Al2O3 deposition trimethylaluminum (TMA) (99.999+% Al) and deionized H2O precursors were used. For the HfO2 deposition Hf[N(CH3)2]4 (98+%) and deionized H2O precursors were used. Only Hf[N(CH3)2]4 was heated to 70 °C to achieve higher vapor pressure. N2 gas of purity 6.0 was used as carrier gas for the precursors. Prior to FZP deposition, growth per cycles (GPC) of the Al2O3 and HfO2 thin films were measured in a test deposition on Si Wafers manufactured via a spectroscopic ellipsometry and determined to be 0.078 and 0.097 nm per cycle, respectively. For the ML‐FZP nonstandard 1:2 line (HfO2) to space (Al2O3) ratio was used instead of the standard 1:1, that allowed for faster deposition. The N2 flow, chamber pressures and pulse and purge times are listed in TableALD for Tapered ML‐FZPs: Al2O3–SiO2 multilayers of 1:1 line to space ratio were deposited on tapered micropillars prepared by PFIB (see Micropillar Fabrication via PFIB) using a SENTECH SI ALD LL instrument (Germany) at 200 °C. For the Al2O3 deposition TMA (99.999+% Al) and deionized H2O precursors were used. For the SiO2 deposition bis[diethylamino]silane (BDEAS) and O2 plasma (O2 gas of purity 6.0) were used. The plasma power was 200 W. Only BDEAS was heated to 70 °C to achieve the necessary vapor pressure. N2 gas of purity 6.0 was used as carrier gas for the precursors. Prior to FZP deposition, GPC of the Al2O3 and SiO2 thin films were measured in a test deposition on Si Wafers via a spectroscopic ellipsometry and determined to be 0.059 and 0.1304 nm per cycle, respectively. The N2 flow, chamber pressures and pulse and purge times are listed in TableFIB Slicing, Transfer, and Polishing of the ML‐FZP: The deposited glass fiber was sliced and polished with a Ga+ ion beam in the FEI Nova NanoLab 600 dual‐beam instrument. The transfer of the multilayer slice on to a Mo lift‐out grid was performed by a micromanipulator in the dual‐beam instrument. A comment on FIB induced damage can be found in the Supporting Information.Micropillar Fabrication via PFIB: Tapered micropillars were fabricated from single crystalline samples of Si(100) and Au(111)  by using a ThermoScientific Helios Plasma FIB. iFast recipes were created for controlling the pillar milling processes automatically. In order to fabricate pillars with reasonable throughput, high FIB beam currents for milling are selected. To achieve a lower taper angle of the pillars, multiple concentric angular milling ring patterns were generated by iFast recipe with outer diameters dynamically reducing in a pre‐defined step size during the pillar fabrications. The fabrication parameters are summarized in TableWavelength‐Dispersive X‐Ray Spectroscopy: Al2O3 and HfO2 thin films were deposited via ALD on separate 4 in. Si wafers using the ALD parameters of Table  2O3 and HfO2 thin films were measured using an accelerating voltage of 5 kV, a beam current of 40 nA and a beam diameter of 5 µm. To eliminate the instrument effects an analysis with matched standards was performed. A pure sapphire mineral was used as a standard for the elements Al and O, and a Hf crystal was used as a standard for the element Hf. A background correction was done by measuring the mean intensity of the background radiation on both sides of the peak and subtracting this value from the intensity of the element characteristic X‐rays. To account for the matrix effects, a ZAF  correction was done in the WDX software. An iterative procedure was followed using the K‐ratios for calculating the stoichiometry.X‐Ray Photoelectron Spectroscopy: Al2O3 and HfO2 thin films were deposited via ALD on separate 4 in. Si wafers using the ALD parameters in the Table  α radiation . Both samples suffered from high C concentration related to surface contaminants. To confirm, the XPS measurements were repeated after an in situ Ar+ sputter‐etching . The XPS analysis with in situ Ar+ sputter‐etching showed <1.5 at% C concentration. The very low C1s peak present at the XPS data of Figure S1b,f (Supporting Information) are related to the surface C–C and C–H contamination during the XPS measurement, which was not visible directly after the Ar+ sputter‐etching. This suggests that the C content in the Al2O3 and HfO2 films should be well below <1.5 at%.Scanning‐ and High‐Resolution Transmission Electron Microscopy: A cross section specimen from a deposited glass–fiber in longitudinal direction , a 100 mm2 JEOL Centurio EDX detector and a Gatan GIF Quantum ERS spectrometer. The microscope was operated at 200 kV, a semiconvergence angle of 21 mrad, giving rise to a probe size of 1 Å for analytical measurements. A collection semiangle of 112 mrad was used for EELS measurements. Figure  a was thiHRTEM experiments were performed at 200 kV with JEOL JEM‐ARM200F, equipped with a cold field‐emission gun and a CETCOR image corrector (CEOS Co. Ltd.).X‐Ray Reflectometry: X‐ray reflectometry measurements were conducted with a Siemens D5005 diffractometer equipped with an X‐ray mirror for Cu Kα radiation on the same samples explained in XPS section. The data were fitted with the LEPTOS (Bruker) software.Synchrotron Measurements: A STXM beamline (MAXYMUS) with elliptical undulator of the APPLE II type (UE46‐PGMII) located at BESSY II was utilized for the synchrotron experiments. For the diffraction efficiency measurements a pinhole of diameter, d = 4 µm was scanned over an area of 85 × 60 µm to image the ML‐FZP mounted on the lift‐out grid as in Figure  Michelson contrast was used for image contrast determination of the BAM sample in Figure  + and Ga+ plasma FIB instruments used in this work.The MPG filed a patent application for the proposed techniques. One of the authors, C.J. works for Thermo Fisher Scientific, which produces the XeSupplementaryClick here for additional data file.