Advanced Microscopy Techniques Research Articles

Application of Advanced Microscopy Techniques to The Characterization of Mixed Matrix Membranes.

Mixed matrix membranes (MMMs) have emerged as promising materials for various separation processes due to their tunable properties, enhanced separation performance and reproducibility. In this review paper, we provide a comprehensive overview of the methodologies, challenges, and applications associated with the characterization of MMMs using two advanced imaging techniques: Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) and Transmission Electron Microscopy (TEM). We begin by outlining the principles and capabilities of FIB-SEM and TEM, emphasizing their suitability for studying the microstructure, morphology, and composition of MMMs at nanoscale resolution. Subsequently, we discuss the specific challenges and limitations encountered in the characterization of MMMs using these techniques, including sample preparation, image acquisition, and data interpretation. Furthermore, we review the diverse applications of FIB-SEM and TEM in elucidating the structure-property relationships of MMMs. Through illustrative examples, we highlight the valuable insights gained from these imaging techniques in optimizing MMMs for various separation applications. Finally, we propose future directions and emerging trends in MMM characterization, including the integration of lasers into FIB-SEM and in situ characterization techniques, to address current challenges and push the boundaries of MMM design and performance.

Correlative analysis of advanced microscopy techniques for metallography and corrosion microstructures of bronze phoenician coins

Correlative microscopy stands out for its ability to concurrently acquire and analyze various data types in a multimodal and multiscale environment, enabling precise localization of specific areas within samples and enhancing the accuracy and relevance of analyses. This approach showed promise in revealing the metallurgical history of ancient coins. This article focused on composition, microstructure, and manufacturing process of Phoenician-Punic copper-based alloy coins (5th–4th century BCE). The four coins studied by correlative light and electron microscopy, μ-Raman spectroscopy, and X-ray Microscopy exhibited notable differences in elemental composition and microstructures. These variations are attributed to their origin from casting, followed by striking, and subsequent recrystallization due to a more intricate corrosion process.

Truncated M13 phage for smart detection of E. coli under dark field

BackgroundThe urgent need for affordable and rapid detection methodologies for foodborne pathogens, particularly Escherichia coli (E. coli), highlights the importance of developing efficient and widely accessible diagnostic systems. Dark field microscopy, although effective, requires specific isolation of the target bacteria which can be hindered by the high cost of producing specialized antibodies. Alternatively, M13 bacteriophage, which naturally targets E. coli, offers a cost-efficient option with well-established techniques for its display and modification. Nevertheless, its filamentous structure with a large length-diameter ratio contributes to nonspecific binding and low separation efficiency, posing significant challenges. Consequently, refining M13 phage methodologies and their integration with advanced microscopy techniques stands as a critical pathway to improve detection specificity and efficiency in food safety diagnostics.MethodsWe employed a dual-plasmid strategy to generate a truncated M13 phage (tM13). This engineered tM13 incorporates two key genetic modifications: a partial mutation at the N-terminus of pIII and biotinylation at the hydrophobic end of pVIII. These alterations enable efficient attachment of tM13 to diverse E. coli strains, facilitating rapid magnetic separation. For detection, we additionally implemented a convolutional neural network (CNN)-based algorithm for precise identification and quantification of bacterial cells using dark field microscopy.ResultsThe results obtained from spike-in and clinical sample analyses demonstrated the accuracy, high sensitivity (with a detection limit of 10 CFU/μL), and time-saving nature (30 min) of our tM13-based immunomagnetic enrichment approach combined with AI-enabled analytics, thereby supporting its potential to facilitate the identification of diverse E. coli strains in complex samples.ConclusionThe study established a rapid and accurate detection strategy for E. coli utilizing truncated M13 phages as capture probes, along with a dark field microscopy detection platform that integrates an image processing model and convolutional neural network.Graphical

Utilization of Advanced Microscopy Techniques and Energy-dispersive X-ray Spectroscopy to Characterize Three Piper Species Related to Kava.

Macro- and microscopic techniques have long been used to describe plant materials and establish plant structural profiles. These techniques are commonly used in botanical authentication to identify the genuine and closely allied species used in botanical research. Advanced microscopic techniques were used in this study to differentiate three different Piper species used as kava or kava-kava. The genuine species is Piper methysticum and the other two species commonly called false-kava or kava-kava, are Piper auritum and Piper excelsum. Macroscopic characteristics, including a black-spotted stem and fibrous root, are characteristic of P. methysticum, whereas the stem of P. auritum is greenish with no spots, and the P. excelsum stem is purple-pink. Microscopic attributes include the characteristic collenchyma of stems and the pattern of arrangement of peripheral and medullary vascular bundles. The starch grains are smaller in P. excelsum than in the other two species. Energy-dispersive X-ray spectroscopy analysis of the crystals indicates the expected calcium, magnesium, and silica, along with lesser amounts of sodium, and potassium. The crystals present in the Piper species vary in shape, size, and elemental composition. Combining macro- and microscopical techniques and resulting characteristics are instrumental in differentiating the three Piper species.

Advanced Electron Microscopy Characterisation of Aluminium-Oxygen Coordination in Catalyst Supports

Performance of alumina-supported catalyst systems are significantly affected by how the tetrahedral (Al-O4) and octahedral (Al-O6) coordination are mixed and blended with each other. Characterisation of aluminium-oxygen coordination is thus important to understand how catalysts interact with alumina at the microscopic level. Here we report the application of two advanced electron microscopy techniques on aluminium-oxygen coordination. The first technique is electron energy loss spectroscopy (EELS) that can reveal detailed electronic structure profile of aluminium valence band to analyse how aluminium is coordinated by oxygen. The second technique is pair distribution function (PDF) based on electron diffraction (ED), employed to measure aluminium-oxygen bond lengths associated with the coordination geometry.

Effects of non-isothermal aging and microalloying on precipitation strengthening for Al-Mg-Si alloys

The impacts of non-isothermal aging (NIA) and microalloying (Ag and/or Cu) on precipitation strengthening and mechanical properties in Al-Mg-Si alloys are investigated using advanced microscopic analysis techniques combined with tensile testing. The NIA treatment exhibits different age hardening for this alloy with different compositions. The Ag-Cu-added A3 alloy exhibits the most significant enhancement in peak hardness under NIA treatment than that of the isothermal aging (IA), while only marginal increases are observed for the other alloys (the base A1 alloy and the Cu/Ag-added alloys). The optimal NIA treatment for the Ag-Cu-added A3 alloy is identified as follows: 200 oC × 1h + cooling down to 180 oC at 20oC/h. This specific treatment leads to a peak hardness of 139.0 HV and an ultimate tensile strength of 384.0MPa for this alloy, which is 16.0 HV higher than that of the base Al-Mg-Si alloy when subjected to IA at 180oC. These findings provide valuable insights for the advancement of novel aluminum alloys and heat treatment methodologies.

AI-Based solutions for current challenges in regenerative medicine

The emergence of Artificial Intelligence (AI) and its usage in regenerative medicine represents a significant opportunity that holds the promise of tackling critical challenges and improving therapeutic outcomes. This article examines the ways in which AI, including machine learning and data fusion techniques, can contribute to regenerative medicine, particularly in gene therapy, stem cell therapy, and tissue engineering. In gene therapy, AI tools can boost the accuracy and safety of treatments by analyzing extensive genomic datasets to target and modify genetic material in a precise manner. In cell therapy, AI improves the characterization and optimization of cell products like mesenchymal stem cells (MSCs) by predicting their function and potency. Additionally, AI enhances advanced microscopy techniques, enabling accurate, non-invasive and quantitative analyses of live cell cultures. AI enhances tissue engineering by optimizing biomaterial and scaffold designs, predicting interactions with tissues, and streamlining development. This leads to faster and more cost-effective innovations by decreasing trial and error. The convergence of AI and regenerative medicine holds great transformative potential, promising effective treatments and innovative therapeutic strategies. This review highlights the importance of interdisciplinary collaboration and the continued integration of AI-based technologies, such as data fusion methods, to overcome current challenges and advance regenerative medicine.

The Lattice Strain Distribution in GexSn1-x Micro-Disks Investigated at the Sub 100-nm Scale

Experimental assessment of the strain tensor within a microstructure is challenging, especially for small mechanical deformations acting over submicron length scales. In this work, we fully characterize the spatial strain distribution within a suspended micro-disk laser made of Ge1-xSnx alloy, with fine resolution <200 nm. We employ Scanning X-ray Diffraction Microscopy, a model-free method based on synchrotron radiation, to directly obtain maps of all components of lattice strain and rotation, including the shear strains, finding them on a magnitude ~10-3. We correlate these small elastic deformations to structural defects and the relaxation of the three-dimensional microstructure, demonstrating the potential of an advanced X-ray microscopy technique for microelectronics.

The Alphabet Soup of Microscopy: An Introduction to Advanced Imaging Techniques. Part II: What the FLIP, FLAP, FRAP, FRET, and FLIM is Going On?

Abstract The use of fluorescence in imaging has been a pivotal factor in advancing our scientific understanding. As microscopy continues to evolve, the terminology used to describe these techniques becomes increasingly complex, often resulting in a bewildering array of acronyms that resemble alphabet soup. Among the most prominent acronyms are those associated with advanced fluorescence microscopy techniques: Fluorescence Loss in Photobleaching (FLIP), Fluorescence Localization after Photobleaching (FLAP), Fluorescence Recovery after Photobleaching (FRAP), Förster Resonance Energy Transfer (FRET), and Fluorescence Lifetime Imaging Microscopy (FLIM). Each of these methods provides invaluable insights into molecular dynamics and interactions within living cells and tissues. This article, Part 2 of the Alphabet Soup of Microscopy series, aims to clarify these widely used fluorescence microscopy techniques and to illuminate their contributions to our understanding of cellular processes.

Correlative electron microscopy of discontinuous precipitation

This paper presents a comprehensive study on discontinuous precipitation (DP) in Al-22 at.% Zn alloy, leveraging the correlative and synergic effects of advanced microscopy and modelling techniques. The study is structured into three pivotal steps: in-situ scanning electron microscopy combined with an energy backscattered diffraction analysis for real-time monitoring and an analysis of DP reaction kinetics and grain boundary misorientations; transmission electron microscopy and an energy X-ray dispersive microanalysis for detailed observation of solute-depleted α and solute-rich β lamellae, offering insights into the kinetics of the reaction front (RF) and solute concentration profiles; and a sophisticated modelling approach addressing the limitations of direct observation methods by simulating solute concentration changes within αphase lamellae during RF go-and-stop cycles. This multifaceted approach elucidates the nuances of DP mechanisms, from the nucleation and growth of precipitates to the estimation of instantaneous RF velocity and the impact of grain boundary misorientations. The integration of these methodologies enhances our understanding of DP kinetics and morphology at both meso- and nanoscales, highlighting the significant role of correlative microscopy in metallurgical research. The findings not only deepen our grasp of the fundamental mechanisms driving DP reactions but also provide a valuable comparison framework for experimental data, potentially guiding the development and optimization of metallurgical processes.

Taxonomic insights into medicinal plants pollen, using advanced microscopy techniques, from the Western Ghats, India

Pollen morphological study on 18 medicinal plants from the Western Ghats (India) was carried out using light microscopy (LM), confocal laser scanning microscopy (CLSM) and field emission scanning electron microscopy (FESEM) techniques. The principal aim was to obtain high-resolution three-dimensional images and ultra-structure of the pollen grains of the studied taxa. The palyno-morphological characters, such as polar axis, equatorial diameter, P/E ratio, pollen shape, aperture type and number, exine thickness and surface ornamentation will be useful in the identification and taxonomic classification of the studied medicinal plants. Statistical analysis reveals grouping of the taxa in three different clusters and also isolated presence of taxa on the principal component analysis (PCA) biplot, irrespective of their families, showing similarities and/or variations among the studied taxa, belonging to the same and/or different families. The present study has taxonomic significance, providing high taxonomic resolution (identification of the taxa up to species level), and further aid in the identification and delimitation of the 18 studied plants.

Pentatwinned AuAg Nanorattles with Tailored Plasmonic Properties for Near-Infrared Applications.

Noble metal nanoparticles, particularly gold and silver nanoparticles, have garnered significant attention due to their ability to manipulate light at the nanoscale through their localized surface plasmon resonance (LSPR). While their LSPRs below 1100 nm were extensively exploited in a wide range of applications, their potential in the near-infrared (NIR) region, crucial for optical communication and sensing, remains relatively underexplored. One primary reason is likely the limited strategies available to obtain highly stable plasmonic nanoparticles with tailored optical properties in the NIR region. Herein, we synthesized AuAg nanorattles (NRTs) with tailored and narrow plasmonic responses ranging from 1000 to 3000 nm. Additionally, we performed comprehensive characterization, employing advanced electron microscopy and various spectroscopic techniques, coupled with finite difference time domain (FDTD) simulations, to elucidate their optical properties. Notably, we unveiled the main external and internal LSPR modes by combining electron energy-loss spectroscopy (EELS) with surface-enhanced Raman scattering (SERS). Furthermore, we demonstrated through surface-enhanced infrared absorption spectroscopy (SEIRA) that the NRTs can significantly enhance the infrared signals of a model molecule. This study not only reports the synthesis of plasmonic NRTs with tunable LSPRs over the entire NIR range but also demonstrates their potential for NIR sensing and optical communication.

Influence of cooling bed temperature on microstructural, mechanical properties and surface quality of high-strength seismic-resistant rebar

High-strength seismic-resistant rebar is essential in construction engineering, yet balancing its strength, plasticity, seismic resistance, and surface quality poses challenges. This research explores niobium-vanadium-nitrogen composite reinforced steel, examining how cooling bed temperature affects its microstructure, mechanical properties, and surface quality using advanced microscopy techniques. The research reveals that the steel's microstructure primarily consists of ferrite and pearlite. As cooling bed temperatures decrease, both the proportion and colony size of pearlite, along with ferrite size, show a declining trend. Specifically, reducing the cooling bed temperature from 1100 °C to 900 °C results in a decrease in yield strength, tensile strength, and tensile-to-yield strength ratio by 6 MPa, 27 MPa, and 0.03, respectively. Meanwhile, elongation after fracture, total elongation at maximum force, and strength-ductility balance improve by 3.8%, 1.5%, and 1.93%, respectively. At a cooling bed temperature of 1100 °C, significant blistering occurs on the steel surface. Significant blistering at 1100 °C vanishes at 1000 °C and 900 °C, with diminished silicon enrichment in the surface oxide scale. A cooling bed temperature of 1000 °C ± 20 °C is recommended for optimal performance. This study provides a temperature control strategy for the properties of high-strength seismic-resistant rebar rated at 650 MPa and above, contributing valuable insights to the field of advanced construction materials.

Enhancing plasticity in BCC Mg-Li-Al alloys through controlled precipitation at grain boundaries

This study investigates the improvement of plasticity in body-centered cubic magnesium (Mg)-lithium (Li)-aluminum (Al) alloys, crucial for lightweight structural applications. The ternary Mg-Li-Al alloys exhibits high strength but low ductility. Precipitates at grain boundaries in these alloys, linked to reduced plasticity, are examined for their crystal structure and composition. Advanced microscopic techniques reveal the transformation of precipitates and the development of specific structures at grain boundaries. Thermodynamics of element diffusion at grain boundaries are explored through first-principles calculations, and a phase-field simulation models precipitate evolution. Molecular dynamics simulations elucidate nanoscale mechanisms governing the transition from brittle to ductile fracture modes during artificial aging. The D03−Mg3Al at grain boundaries is a brittle phase, and through a 170 °C aging treatment, it induces the precipitation of lamellar α-Mg phase with D03−Mg3Al as nucleation sites. The occupancy energy of Al atoms at Li sites in α-Mg is found to be lower than that in D03−Mg3Al, leading to the dissolution of D03−Mg3Al. The α-Mg, characterized by a stronger metallic nature, exhibits a better-matched modulus with the matrix and enhanced dislocation mobility. The precipitation of α-Mg plays a pivotal role in significantly improving the ductility of the alloy. This work contributes to the understanding of the complex interplay between alloy composition, grain boundary precipitates, and plasticity, as well as brings insights to guide interfacial control in the development of advanced Mg-Li-Al alloys for structural applications.

Investigation of AlGaN UV emitting tunnel junction LED devices by off-axis electron holography

Here we use off-axis electron holography combined with advanced transmission electron microscopy techniques to understand the opto-electronic properties of AlGaN tunnel junction (TJ)-light-emitting diode (LED) devices for ultraviolet emission. Four identical AlGaN LED devices emitting at 290 nm have been grown by metal–organic chemical vapour deposition. Then Ge doped n-type regions with and without InGaN or GaN interlayers (IL) have been grown by molecular beam epitaxy onto the top Mg doped p-type layer to form a TJ and hence a high quality ohmic metal contact. Off-axis electron holography has then been used to demonstrate a reduction in the width of the TJ from 9.5 to 4.1 nm when an InGaN IL is used. As such we demonstrate that off-axis electron holography can be used to reproducibly measure nm-scale changes in electrostatic potential in highly defected and challenging materials such as AlGaN and that systematic studies of devices can be performed. The LED devices are then characterized using standard opto-electric techniques and the improvements in the performance of the LEDs are correlated with the electron holography results.

Influence of Pristine NMC811 Secondary and Primary Particle Surfaces on the Materials Reactivity

Vehicle electrification currently relies on lithium-ion batteries using nickel, manganese and cobalt (NMC)-based lamellar oxides as positive electrode materials. In order to achieve higher energy densities, the market is now moving towards NMC with high nickel content. Nevertheless, these materials have been widely demonstrated to suffer from serious off-gassing problems that reduce cycle life and pose safety concerns. Not only gases resulting from the formation of SEI at the negative electrode, but the production of O2, CO and CO2 have also been noted by different research groups.1–3 Despite the large number of studies analyzing the material during and after cycling, we noted a lack of in-depth characterization of the surface of Nickel-rich NMCs in their initial state. The need for such information is increasingly necessary with the recent development of new technologies for surface modification such as coating to limit the instability of high Nickel content NMCs.4,5 The surface reactivity of Ni-rich layered transition metal oxides is instrumental to the performance of batteries based on these positive electrode materials. Most often, strong surface modifications are detailed with respect to a supposed ideal initial state. The study the LiNi0.8Mn0.1Co0.1O2 (NMC811) cathode material in its pristine state, hence before any contact with electrolyte or cycling, was performed thanks to advanced microscopy and spectroscopy techniques in order to fully characterize its surface down to the nanometer scale. Scanning transmission-electron microscopy-electron energy-loss spectroscopy (STEM-EELS), solid state nuclear-magnetic-resonance (SS-NMR) and X-rays photoelectron spectroscopy (XPS) are combined and correlated in an innovative manner.Specifically, using a high magnetic field NMR spectrometer, it was possible to evaluate the total amount of diamagnetic Li in the samples. Thanks to XPS it was then possible to identify the lithiated species present on the surface and to use such information to better deconvolve the NMR spectra. Li2O, Li2SO4 and LiOH were identified as the main contaminants on the surface of NMC in its initial state. On the other hand, the quantity of Li2CO3 initially estimated by XPS could be substantially re-evaluated using NMR (54% by XPS vs. only 1% by NMR). These two techniques, providing only partial and often imprecise information when used separately, have proven to be fundamental instruments in the analysis of surface species when used in a complementary manner.The results demonstrate that in usual storage conditions after synthesis, the extreme surface is already chemically different from nominal values. In particular, nickel is found in a reduced state compared to the bulk value and a Mn enrichment is determined in the first few nanometers of primary particles. Further exposition to humid air allows for quantifying the formed lithiated species per gram of active material, identifying their repartition and proposing a reaction path in relation with the instability of the surface. Modifying synthesis conditions impacts noticeably the previously identified surface particularities and, consequently, the materials reactivity.References :(1) Jung, R.; Metzger, M.; Maglia, F.; Stinner, C.; Gasteiger, H. A. Oxygen Release and Its Effect on the Cycling Stability of LiNixMnyCozO2 (NMC) Cathode Materials for Li-Ion Batteries. Journal of The Electrochemical Society 2017, 164 (7), A1361–A1377. https://doi.org/10.1149/2.0021707jes.(2) Renfrew, S. E.; McCloskey, B. D. Residual Lithium Carbonate Predominantly Accounts for First Cycle CO2 and CO Outgassing of Li-Stoichiometric and Li-Rich Layered Transition-Metal Oxides. J. Am. Chem. Soc. 2017, 139 (49), 17853–17860. https://doi.org/10.1021/jacs.7b08461.(3) Renfrew, S. E.; McCloskey, B. D. The Role of Electrolyte in the First-Cycle Transformations of LiNi 0.6 Mn 0.2 Co 0.2 O 2. J. Electrochem. Soc. 2019, 166 (13), A2762–A2768. https://doi.org/10.1149/2.1561912jes.(4) Zhang, H.; Liu, H.; Piper, L. F. J.; Whittingham, M. S.; Zhou, G. Oxygen Loss in Layered Oxide Cathodes for Li-Ion Batteries: Mechanisms, Effects, and Mitigation. Chem. Rev. 2022, acs.chemrev.1c00327. https://doi.org/10.1021/acs.chemrev.1c00327.(5) Manthiram, A.; Song, B.; Li, W. A Perspective on Nickel-Rich Layered Oxide Cathodes for Lithium-Ion Batteries. Energy Storage Materials 2017, 6, 125–139. https://doi.org/10.1016/j.ensm.2016.10.007. Figure 1

A HRTEM-EDS-SAED Study of Precipitates in Mg-Zn-Zr Alloy

In this study, aging heat treatment was performed on extruded Mg-Zn-Zr alloy. Aging was performed on as-extruded samples at 180°C for 1-2-4-8 and 24 hours. Microhardness tests were made to obtain the aging curve. Detailed transmission electron microscopy (TEM) examinations such as selected area electron diffraction (SAED), energy dispersive spectrometry (EDS), and high resolution transmission electron microscopy (HRTEM) were used to investigate the microstructure, second phases, and precipitates. Size, type, and morphologies of second phases and precipitates were studied aided by advanced transmission electron microscopy techniques. With the help of hardness test results peak aged condition was found to be 8 hour aging. TEM examinations were detailed for the 8-hour aged sample. It was observed that the precipitates responsible for age hardening were MgZn2. Moreover, the majority of these precipitates were β’1 phase.

The evolution of organic material on Asteroid 162173 Ryugu and its delivery to Earth

The recent return of samples from asteroid 162173 Ryugu provides a first insight into early Solar System prebiotic evolution from known planetary bodies. Ryugu’s samples are CI chondrite-like, rich in water and organic material, and primarily composed of phyllosilicate. This phyllosilicate surrounds micron to submicron macromolecular organic particles known as insoluble organic matter. Using advanced microscopy techniques on Hayabusa-2 samples, we find that aqueous alteration on Ryugu produced organic particles richer in aromatics compared to less altered carbonaceous chondrites. This challenges the view that aromatic-rich organic matter formed pre-accretion. Additionally, widespread diffuse organic material occurs in phyllosilicate more aliphatic-, carboxylic-rich, and aromatic-poor than the discrete organic particles, likely preserving the soluble organic material. Some organic particles evolved to encapsulate phyllosilicate, indicating that aqueous alteration on Ryugu led to the containment of soluble organic matter within these particles. Earth therefore has been, and continues to be, delivered micron-sized polymeric organic objects containing biologically relevant molecules.

High-speed imaging and quantitative analysis of nonequilibrium stochastic processes using atomic resolution electron microscopy

Abstract Chemical phenomena are inherently complex and stochastic, making them difficult to fully understand using conventional ensemble-averaged analytical methods. These methods primarily capture long-lived species and common structural features, limiting the study of transient intermediates and minute structural characteristics. In contrast, single-molecule time-resolved analysis using advanced microscopy techniques, particularly transmission electron microscopy, offers high spatial and temporal resolution to observe the nonequilibrium dynamics of molecules and their assemblies. This account discusses the authors’ research on developing transmission electron microscopy techniques to visualize intricate and transient interactions within molecular systems, enhancing the understanding of chemical phenomena at atomic and molecular levels.

An atlas of the tomato epigenome reveals that KRYPTONITE shapes TAD-like boundaries through the control of H3K9ac distribution

In recent years, the exploration of genome three-dimensional (3D) conformation has yielded profound insights into the regulation of gene expression and cellular functions in both animals and plants. While animals exhibit a characteristic genome topology defined by topologically associating domains (TADs), plants display similar features with a more diverse conformation across species. Employing advanced high-throughput sequencing and microscopy techniques, we investigated the landscape of 26 histone modifications and RNA polymerase II distribution in tomato (Solanum lycopersicum). Our study unveiled a rich and nuanced epigenetic landscape, shedding light on distinct chromatin states associated with heterochromatin formation and gene silencing. Moreover, we elucidated the intricate interplay between these chromatin states and the overall topology of the genome. Employing a genetic approach, we delved into the role of the histone modification H3K9ac in genome topology. Notably, our investigation revealed that the ectopic deposition of this chromatin mark triggered a reorganization of the 3D chromatin structure, defining different TAD-like borders. Our work emphasizes the critical role of H3K9ac in shaping the topology of the tomato genome, providing valuable insights into the epigenetic landscape of this agriculturally significant crop species.