Electron Image Analysis Research Articles

Scale dependence of textural alignment in shales quantified using electron microscopy

Textural alignment is the primary cause of anisotropy associated with the permeability and mechanical and acoustic properties of rocks. Although particle alignment in shales is known to be influenced by burial depth, mineralogical composition, and diagenetic history, understanding how these influence image-based measures of compaction at the micrometer to nanometer scale is not well understood. Here, we use a novel method, which combines scanning electron microscopy and image analysis algorithms, to quantify clay alignment in sedimentary rocks. For a given field of view, the alignment intensity in shales from the Podhale Basin in Poland is positively correlated with clay content based on image analysis. However, our analysis also shows that the intensity is strongly dependent on the spatial scale of imaging: as magnification increases and the field of view in the SEM images decreases, the alignment intensity drops. Therefore, in images with small fields of view, clay alignment is determined by the presence of localized equant silicate mineral grains, which effectively hinder the alignment process at the micrometer scale. By contrast, in images with larger fields of view, alignment intensity is determined by the overall compaction state. We also show that when the field of view is constant, alignment intensity decreases with pixel density, and we suggest a range of operating parameters required for obtaining optimum values for alignment intensity from SEM images of shales.

Changes in creep property and precipitates due to aging of T91 steel after long-term service

The creep deformation behavior of virgin and long-term service-aged (77000 h) T91 steel of superheater tubes in a power plant was assessed via creep tests along with microstructure examination. A series of miniature tensile creep tests were conducted at 590 °C under applied stresses ranging from 120.0 to 224.1 MPa for both material types. The creep behavior was examined in terms of variations in the microstructural properties correlated with creep rupture life and ductility. X-ray diffraction, optical microscopy, field-emission scanning electron microscopy, hardness measurements, and image analyses were used to analyze the shape and morphology of the existing δ-ferrite, tempered martensite microstructure, and precipitates in both the virgin and long-term service-aged materials. The presence of larger residual δ-ferrite in the virgin microstructure was one of the causes of creep rupture strength reduction in the virgin material. On the contrary, the synergistic effect of the tempered martensite lath microstructure and additional precipitation strengthening of fine precipitates had beneficial effects translating to retention of the creep rupture strength of the long-term service-aged material.

Developing active poly(vinyl alcohol)-based membranes with encapsulated antimicrobial enzymes via electrospinning for food packaging

The present study addresses the immobilization of the GOX enzyme into a conventional polyvinyl alcohol (PVOH) membrane and a multi-layer membrane system (PCL/PVOH/PCL), both synthetized through electrospinning technique. More specifically, the enzyme was encapsulated inside a PVOH hydrophilic membrane, which was in turn protected by two hydrophobic polycaprolactone (PCL) membranes (multilayer system). The manufactured systems were characterized through IR spectroscopy, enzymatic kinetics and mechanical properties. Moreover, the morphology of the fibers composing the membranes was studied by Scanning Electronic Microscopy (SEM) and image analysis. Finally, the enzyme activity of the systems was in-vitro bacterial cultures using E. coli. The results obtained indicate that the addition of the GOX enzyme involves a slight reduction of the mechanical properties (maximum strain at break and maximum stress), which is associated with an increase of the polydispersity of the generated fibers. However, the mats showed antibacterial properties, which were evidenced by the inhibition of E. coli growth. These results indicate that this technique is suitable for the immobilization of enzymes with potential use in the active packaging industry.

Role of ITZ in the Degradation Process of Blended Cement Concrete under Magnesium Sulfate Attack

Although the importance of the interfacial transition zone (ITZ) to sulfate attack is obvious, the relation between the ITZ and the rate of degradation is still unclear. The impact of ITZ volume content on the degradation of concrete, including length, mass, and elastic dynamic modulus (Ed) variation, fully immersed in magnesium sulfate solution were investigated in this work. The microstructure was analyzed by backscattered electron imaging (BSE) and energy dispersive X-ray spectroscopy (EDX) mapping analysis. The performance of concrete exposed to magnesium sulfate highly depended on the aggregate and the composition of raw materials. There was a trend toward more serious deterioration with increasing ITZ in the reference and limestone filler blended group after 12 months of exposure. The degradation of specimens made with slag was independent of the variation of ITZ content. Gypsum tended to precipitate in the ITZ under magnesium sulfate attack. The EDX analysis confirmed the decomposition of C-S-H, accompanied by the formation of degradation products M-S-H.

Effects of deep cryogenic treatment on microstructure, impact toughness and wear resistance of an AISI D6 tool steel

Several studies have shown that cryogenic treatments improve many properties of tool steels. Basically the cryogenic treatment reduces the amount of retained austenite and promotes ultrathin carbides precipitation. In this study, samples of AISI D6 were submitted to several heat treatment cycles, aiming to analyze the influence of cryogenic treatment on microstructure, impact toughness and wear resistance. These cycles were quenching and tempering, quenching and tempering followed by deep cryogenic treatment (2 and 24 h) and, finally, quenching and tempering followed by deep cryogenic treatment plus double tempering. By the comparison with other studies and by the application of techniques such as Electron Microscopy, Image Analysis and X-ray Diffraction, it was observed that the hardening step (austenitization time and temperature) plays a fundamental role in the retained austenite level and in the cryogenic treatment results. The thermal cycles act both on the martensite formed directly from the quenching (primary martensite) and on the martensite formed from the retained austenite. It was concluded that cryogenic treatment followed by double tempering produced a reduction in the amount of retained austenite and consequently leads to secondary carbides precipitation. This fact has a marked role on the mechanical properties, keeping the wear values unchanged for the studied conditions.

Influence of strain rate and temperature on the deformation mechanisms of a fine-grained Ti-6Al-4V alloy

Depending on their initial microstructure, titanium alloys, as the Ti-6Al-4V may have activation of different deformation mechanisms during hot forming processes. In this work, interrupted tensile tests and heat treatments are used to improve the understanding of the mechanical and microstructural behaviour of a fine-grained Ti-6Al-4V alloy at two temperatures (750°C and 920°C) and so for two different β phase fractions. The microstructural features like, α grain size and phase fraction, were determined by Scanning Electron Microscope (SEM) and image analysis. Moreover, evolution of the preferred crystallographic orientation of α grains and local misorientations between and inside grains were obtained by Electron Backscatter Diffraction (EBSD). The strain rate sensitivity parameter as well as the activation energy were deduced from mechanical tests. It appears, from all these microstructural and mechanical data, that several mechanisms are activated depending on the strain level and on the temperature range. At 750°C, for a high strain rate, the deformation is mainly controlled by dislocations activity in the α phase (texture changes, dynamic recrystallization) and, at very low strain rate, by probably GBS accommodated with dislocations activity into α (recovery) and β. On the contrary at 920°C, a clear decrease of the overall texture intensity associated with a high m value suggests that GBS is the dominant mode of deformation. Nevertheless, as the α∕β volume fraction is around 48%/52% at this temperature, not only the α phase but also the β phase as well as α∕α and β∕β boundaries might contribute to the flow behaviour. During long deformation time (low strain rate and high temperature), dynamic coarsening behaviour (both into α and β), that is controlled by bulk diffusion, can occur and modify the type, the distribution and decrease the number of α∕β, α∕α and β∕β boundaries. This can be partly related to the flow hardening observed at 920°C and 10−4s−1.

Investigation of Pore Structures in Shallow Longmaxi Shale, South China, via Large-Area Electron Imaging and Neutron Scattering Techniques

Pore structures are significant for evaluating shale gas production since they not only control gas capacity but also affect hydraulic fracturing. In this study, we investigated the pore structures of Longmaxi (LMX) shale samples from a shallow drill core in Guizhou Province, China, with large-area electron imaging and neutron scattering. Pores related to the mineral matrix and organic matter (OM) were identified, and the former had greater impacts on the pore shape and porosity, which would enhance fracturing. Large-area electron imaging analysis (with 15 nm resolution) indicated that the majority of LMX shale porosity was contributed by nanopores with diameters smaller than 400 nm. According to the neutron scattering data, micropores and mesopores dominated the pore size distribution (PSD), and pores within cross sections cut perpendicular to the bedding showed preferential orientation accompanied by larger porosity. The porosity (4.6–5.6%) calculated from neutron scattering data was approximately 2 times larger than that calculated from imaging data, suggesting that nanopores smaller than 15 nm contributed more to the porosity than those larger than 100 nm, as they were more resistant to compaction. This study indicated that pore types were controlled by minerals as well as thermal maturity, while PSDs were affected by tectonic movements. The majority of porosity originated from micropores and mesopores, which confirmed the great potential for shale gas reserves in the LMX shale. This conclusion is helpful for building a shale porosity model and eventually improving the reserve assessment.

In situ liquid SEM imaging analysis revealing particle dispersity in aqueous solutions.

A quantitative description on dispersity of boehmite (γ-AlOOH) particles, a key component for waste slurry at Hanford sites, can provide useful knowledge for understanding various physicochemical nature of the waste. In situ liquid scanning electron microscopy (SEM) was used to evaluate the dispersity of particles in aqueous conditions using a microfluidic sample holder, System for Analysis at Liquid Vacuum Interface (SALVI). Secondary electron (SE) images and image analyses were performed to determine particle centroid locations and the distance to the nearest neighbour particle centroid, providing reliable rescaled interparticle distances as a function of ionic strength in acidic and basic conditions. Our finding of the particle dispersity is consistent with physical insights from corresponding particle interactions under physicochemical conditions, demonstrating delicate changes in dispersity of boehmite particles based on novel in situ liquid SEM imaging and analysis. LAY DESCRIPTION: In situ liquid scanning electron microscopy (SEM) was used to determine the interparticle distance of boehmite (γ-AlOOH) particles, a key component for waste slurry at Hanford sites. This type of quantitative measurement is important to understand various physicochemical nature of the radiological waste containing boehmite. In situ liquid SEM was enabled by a unique vacuum compatible microfluidic cell, System for Analysis at Liquid Vacuum Interface (SALVI). We collected secondary electron (SE) images and performed image analyses to determine particle centroid locations and the distance to the nearest neighbour particle centroid to arrive at the interparticle distances in acidic and basic conditions. Our results show that delicate changes occur among boehmite particles under different pH conditions using novel in situ SEM imaging.

Fine-Scale Heterogeneity of Pyrite and Organics within Mudrocks: Scanning Electron Microscopy and Image Analysis at the Large Scale

Mudrocks are highly heterogeneous in terms of both composition and fabric, with heterogeneities occurring at the submicron to centimetre plus scale. Such heterogeneities are relatively easy to visualise at the micron-scale through the use of modern scanning electron microscopy (SEM) techniques, but due to their inherent fine grain size, can be difficult to place within the greater context of the mudrock as a whole, or to visualise variation when viewed at a centimetre scale. The utilisation of SEM to collect automated high-resolution backscattered (BSE) images (tiles) over whole, polished thin-sections presents a potential large data bank on compositional and fabric changes that can be further processed using simple image analysis techniques to extract data on compositional variation. This can then be plotted graphically in 2D as colour contoured distribution maps to illustrate any observed variability. This method enables the easy visualisation of micron-scale heterogeneity present in mudrock, which are here illustrated and discussed for pyrite and organic content at the larger (thin-section) centimetre scale. This does not require the use of other techniques such as energy dispersive x-ray (EDX) mapping to identify phases present, but instead utilizes BSE images that may already have been collected for textural fabric studies. The technique can also be applied to other phases in mudrocks, such as carbonates and silicates, as well as porosity. Data can also be extracted and used in a similar fashion to bulk compositional analytical techniques such as inductively coupled plasma-atomic emission microscopy (ICP-AES) and carbon, nitrogen and sulphur (CNS) analysis, for average organic carbon and percentage pyrite.

Process-Geometry Interplay in the Deposition and Microstructural Evolution of 7YSZ Thermal Barrier Coatings by Air Plasma Spray

Air plasma spray (APS) yttria-stabilized zirconia (7YSZ) TBCs are often deposited onto model planar surfaces for microstructural evaluation and testing. Recently, an interest on the implications of both TBC deposition and performance on more complex geometries has developed in the scientific and industrial community, as the microstructures can be substantially different depending on the substrate geometry with concomitant implications on performance (e.g. leading-edge failure). In this study, the implications of processing TBCs on cylindrical substrates as opposed to planar surfaces are explored. Particle diagnostics were conducted to understand differences in coating formation dynamics between planar and cylindrical targets. Scanning Electron Microscopy and Image Analysis were concurrently used to quantitatively define the porosity differences among multiple conditions of spraying and to benchmark the transition of porosity from disk to cylinder. Additionally, rods of different radii of curvature were explored in this study to evaluate the effect of the radius of curvature on the TBC deposit microstructure. Finally, a modified approach to established thermoelastic beam curvature techniques is introduced which can see subtle changes in the mechanical properties of coatings produced at torch traverse conditions similar to how they are produced on a curved surface.

Effects of brass coating and nano-SiO2 coating on steel fiber–matrix interfacial properties of cement-based composite

This study focused on the effects of steel fiber coatings on the interfacial properties of a fiber-reinforced cement-based composite. The impact of a brass coating on the interface between the steel fibers and cement matrix was examined, and the improvement effects of a nano-SiO2 coating and nano-SiO2 powder on both the microstructure and interfacial bond properties were compared. A chemical simulation experiment was designed to analyze the brass coating state in a cement hydration environment. The backscattered electron image analysis (BSE-IA) method was used to conduct quantitative analyses of the calcium hydroxide (CH), porosity, and unhydrated cement (UHC) to investigate the microstructure of the fiber–matrix interfacial transition zone (ITZ). Pull-out tests were conducted to determine the interfacial bond performance, and scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) were used for micro-observation and element analyses. The results indicated that because of the destruction of the coating structure and the aggregation of CH caused by the dezincification of the brass in a cement hydration environment, the brass coating did not improve (but weakened) the interfacial microstructure and bond performance. Adding the nano-SiO2 powder directly was a useful method for achieving microstructure densification and defect reduction in both the ITZ and bulk paste, while the nano-SiO2 coating provided a more targeted improvement in the fiber–matrix ITZ, particularly near the fibers, leading to a more effective enhancement of the bond performance.

Influence of the Fabrication Accuracy of Hot-Embossed PCL Scaffolds on Cell Growths.

Polycaprolactone (PCL) is a biocompatible and biodegradable polymer widely used for the realization of 3D scaffold for tissue engineering applications. The hot embossing technique (HE) allows the obtainment of PCL scaffolds with a regular array of micro pillars on their surface. The main drawback affecting this kind of micro fabrication process is that such structural superficial details can be damaged when detaching the replica from the mold. Therefore, the present study has focused on the optimization of the HE processes through the development of an analytical model for the prediction of the demolding force as a function of temperature. This model allowed calculating the minimum demolding force to obtain regular micropillars without defects. We demonstrated that the results obtained by the analytical model agree with the experimental data. To address the importance of controlling accurately the fabricated microstructures, we seeded on the PCL scaffolds human stromal cell line (HS-5) and monocytic leukemia cell line (THP-1) to evaluate how the presence of regular or deformed pillars affect cells viability. In vitro viability results, scanning electron and fluorescence microscope imaging analysis show that the HS-5 preferentially grows on regular microstructured surfaces, while the THP-1 on irregular microstructured ones.

Molecular and skeletal fingerprints of scleractinian coral biomineralization: From the sea surface to mesophotic depths

Reef-building corals, the major producers of biogenic calcium carbonate, form skeletons in a plethora of morphological forms. Here we studied skeletal modifications of Stylophora pistillata (clade 4) colonies that adapt to increasing depths with decreasing ambient light. The coral show characteristic transitions from spherical morphologies (shallow depths, 5 m deep) to flat and branching geometries (mesophotic depths, 60 m deep). Such changes are typically ascribed to the algal photosymbiont physiological feedback with the coral that host them. We find specific fine-scale skeletal variability in accretion of structure at shallow- and mesophotic depth morphotypes that suggest underlying genomic regulation of biomineralization pathways of the coral host. To explain this, we conducted comparative morphology-based analyses, including optical and electron microscopy, tomography and X-ray diffraction analysis coupled with a comprehensive transcriptomic analysis of S. pistillata. The samples originated from Gulf of Eilat in the Red Sea collected along a depth gradient from shallow to mesophotic depths (5 to 60 m). Additional samples were experimentally transplanted from 5 m to 60 m and from 60 m to 5 m. Interestingly, both morphologically and functionally, transplanted corals partly adapt by exhibiting typical depth-specific properties. In mesophotic depths, we find that the organic matrix fraction is enriched in the coralla, well matching the overrepresentation of transcripts encoding biomineralization “tool-kit" structural extracellularproteins that was observed. These results provide insights into the molecular mechanisms of calcification and skeletal adaptation that repeatedly allowed this coral group to adapt to a range of environments presumably with a rich geological past. Statement of SignificanceUnderstanding the reef coral physiological plasticity under a rapidly changing climate is of crucial importance for the protection of coral reef ecosystems. Most of the reef corals operate near their upper limit of heat tolerance. A possible rescue for some coral species is migration to deeper, cooler mesophotic depths. However, gradually changing environmental parameters (especially light) along the depth gradient pose new adaptative stress on corals with largely unknown influences on the various biological molecular pathways. This work provides a first comprehensive analysis of changes in gene expression, including biomineralization “tool kit” genes, and reports the fine-scale microstructural and crystallographic skeletal details in S. pistillata collected in the Red Sea along a depth gradient spannign 5 to 60 m.

Archaeological human hair in a burial context: Microscopical evaluation of samples from Iași, Romania (18th–19th centuries)

Owing to its resistance to degradation, hair is an important bio‐sample with unique properties used to study post mortem decompositional changes. The present paper is a microscopical approach to preserved human scalp hair discovered at the 18th–19th‐century archaeological site of Iași, Romania. The hair fibre alterations were assessed using optical microscopy, scanning electron microscopy, image analysis and hair histological scores. External alterations showed a statistically significant heterogeneous degradation. Image analysis results of the internal degradation of hairs were correlated to external lesions. The results consider the implication of intrinsic biological factors in the post mortem degradation of human hair preserved in graves with comparable environmental conditions.

BSE‐IA reveals retardation mechanisms of polymer powders on cement hydration

AbstractThe retardation effects of two polymer powders, styrene butadiene rubber and ethylene‐vinyl acetate, on cement hydration and the mechanisms in terms of hydration degree and microstructure characteristics were investigated by the combination of backscattered electron image analysis, scanning electron microscopy, energy dispersive spectrometer, and X‐ray diffraction. Results show that the cement hydration can be significantly retarded by both polymers with the reductions in cement hydration degrees and hydration products thickness. Polymers retard cement hydration by agglomerating the polymer particles on the surfaces of cement grains, forming films in different thickness and bonding the calcium ions in the pore solutions. Consequentially, the formation of C‐S‐H and CH is depressed and the Ca/Si ratios in the hydration product layer are raised. The retardation is enhanced with the increase in polymer content. This work helps understand the retardation effect of polymers on cement hydration in depth, which enables rational development of polymer‐modified cement‐based materials for further applications.

Mechanical and wear properties of Si3N4 reinforced titanium composites

In the present study, Si3N4 reinforced titanium composites were produced by powder metallurgy method. The effect of various percentages of Si3N4 (0-9 wt.%) on the microstructure, density, hardness and compressive strength of titanium (Ti) composites have been investigated. After sintering at 1100oC for 120 min., the mechanical properties were significantly developed up to 3 wt.% Si3N4. The highest hardness and the greatest compressive strength were obtained for 3 wt.% Si3N4 reinforced composites (698.5 HV and 1093 MPa) when compared to pure titanium (414.2 HV and 826 MPa). Si3N4 addition developed the wear properties of composites when compared pure titanium. The lowest wear rate (1.36x10-5, 2.75x10-5 and 5.15x10-5 mm3/Nm for 10 N, 20 N and 30 N) were obtained for 3 wt.% Si3N4 reinforced titanium composite. The scanning electron image, elemental mapping and line analyses confirm the uniform distribution of Si3N4 powder in Ti matrix. Above this ratio, the properties of composites were deteriorated due to agglomeration tendency of Si3N4 powder. There were no in-situ formed second phases of composite structure from the X-ray diffraction analyses.

A Comparative Study on the Influence of Chromium on the Phase Fraction and Elemental Distribution in As-Cast High Chromium Cast Irons: Simulation vs. Experimentation

The excellent abrasion resistance of high chromium cast irons (HCCIs) stems from the dispersion of the hard iron-chromium eutectic carbides. The surrounding matrix on the other hand, provides sufficient mechanical support, improving the resistance to cracking deformation and spalling. Prior knowledge of the microstructural characteristics is imperative to appropriately design subsequent heat treatments, and in this regard, employing computational tools is the current trend. In this work, computational and experimental results were correlated with the aim of validating the usage of MatCalc simulations to predict the eutectic carbide phase fraction and the elemental distribution in two HCCI alloys, in the as-cast condition. Microstructural observations were carried out using optical microscopy and SEM. The chemical composition and fraction of each phase was measured by electron probe microanalysis and image analysis, respectively. In all cases, the values predicted by the pseudo-equilibrium diagrams, computed with MatCalc, were in accordance with the experimentally determined values. Consequently, the results suggest that time and resource intensive experimental procedures can be replaced by simulation techniques to determine the phase fraction and especially, the individual phase compositions in the as-cast state.

Electron Tomography Analysis of Thylakoid Assembly and Fission in Chloroplasts of a Single-Cell C4 plant, Bienertia sinuspersici

Bienertia sinuspersici is a single-cell C4 plant species of which chlorenchyma cells have two distinct groups of chloroplasts spatially segregated in the cytoplasm. The central vacuole encloses most chloroplasts at the cell center and confines the rest of the chloroplasts near the plasma membrane. Young chlorenchyma cells, however, do not have large vacuoles and their chloroplasts are homogenous. Therefore, maturing Bienertia chlorenchyma cells provide a unique opportunity to investigate chloroplast proliferation in the central cluster and the remodeling of chloroplasts that have been displaced by the vacuole to the cell periphery. Chloroplast numbers and sizes increased, more notably, during later stages of maturation than the early stages. Electron tomography analyses indicated that chloroplast enlargement is sustained by thylakoid growth and that invaginations from the inner envelope membrane contributed to thylakoid assembly. Grana stacks acquired more layers, differentiating them from stroma thylakoids as central chloroplasts matured. In peripheral chloroplasts, however, grana stacks stretched out to a degree that the distinction between grana stacks and stroma thylakoids was obscured. In central chloroplasts undergoing division, thylakoids inside the cleavage furrow were kinked and severed. Grana stacks in the division zone were disrupted, and large complexes in their membranes were dislocated, suggesting the existence of a thylakoid fission machinery.

Anatomical and Morphological Properties of Trichomes in Four Iranian Native Salvia Species under Cultivated Conditions

The morphological specifications play a substantial role in classification and breeding programs of various plant taxa. In the current study, some macro- and micro-morphological features of Salvia nemorosa, Salvia syriaca, Salvia frigida and Salvia virgata (three accessions), were investigated using scanning electron microscopy and image analysis program. The completely randomized design (CRD) was used to compare the morphological properties (r=4). Macro-morphological and agronomical measurements showed that Salvia species have great potential for cultivation as medicinal and ornamental plants. The cluster analysis suggested that S. virgata and S. frigida located in the same group; on the other side, S. syriaca and S. nemorosa had more similarity. The micro-morphological results showed that glandular trichomes (GTs) and non-glandular trichomes (NGTs) were widely distributed over the leaf and flower surfaces. The peltate GTs with high-density covered sepal and petal and were observed with low density on the leaves. The comparison with different species showed that the highest size and density of GTs belonged to Salvia virgata. The GTs are large and cover a significant portion of the flower surface (3.85-18.45%). The multicellular GTs were observed in S. nemorosa and S. virgata A1 and A2. The uniseriate and multicellular NGTs covered the adaxial leaf surface in different species. Besides, relative variation in GTs was observed in the three studied S. virgata accessions (15.33- 30.58 trichomes per mm2), which indicates the role of environmental conditions in trichomes formation. A wide range of variation in most of the micro- and macro-morphological traits of Salvia genotypes observed in cultivated condition.

Organelle biogenesis and function in plants

All eukaryotic cells contain an endomembrane system with functionally distinct membrane-bound organelles, including the endoplasmic reticulum (ER), Golgi apparatus, trans-Golgi network (TGN), prevacuolar compartment or multivesicular body (PVC/MVB) and vacuole or lysosome. These organelles are generated accurately and efficiently in a well-organized way so as to exert their essential functions in protein transport, plant growth and responses to the environment. Since the year 2000, great progress has been achieved by the Chinese scientists toward our understanding on organelle biogenesis and function in plants. In this review, representative research findings in plant protein trafficking contributed by Chinese scientists are first introduced. Due to the functional importance of PVCs/MVBs, we then focus on the identification and characterization of PVCs/MVBs as well as the mechanisms underlying their biogenesis as regulated by the ESCRT (endosomal sorting complexes required for transport) machinery, especially by the plant-unique ESCRT component, FREE1 (FYVE domain protein required for endosomal sorting 1). In addition, some Chinese scientists recently perform a whole-cell electron tomography analysis in the endomembrane system of root cells to first propose a new MVB-derived vacuole biogenesis model for future investigation of the underlying mechanisms of vacuole formation. Finally, we summarize current research progress on organelle (PVC/MVB and vacuole) biogenesis and function in plants and present future perspectives in this research field.