Transmission Electron Tomography Research Articles

Characterization and the physical properties of nano-sized Bi2O3/polymer for energy and high-refractive index applications

The present work focuses on developing nanocomposites for optoelectronics and energy storage devices leveraging the unique physicochemical features of bismuth (III) oxide (Bi2O3) using a polyvinyl pyrrolidone/polyvinyl alcohol matrix (PV(A/P)). Bi2O3 nanofillers (NF) and Bi2O3/PV(A/P) nanocomposite (NC) films were prepared by facile chemical solution approaches. The X-ray (XRD) data and a transmission electron image revealed that the NF exhibits a monoclinic phase, resembling slightly deformed agglomerated spheres. In the FTIR spectrum, different stretching vibrations of Bi-O-Bi bonds were found, along with their complexation and interaction with the reactive groups in the matrix. The SEM revealed a uniform distribution of Bi2O3 NF loading up to 1.5 wt% on the sample’s surface. Thermal analyses (TGA and DSC thermograms) were used to study the thermal stability, melting, and decomposition temperatures of the NC films. The dielectric features were studied under frequencies between 2 × 103 and 8 × 106 Hz and heating from 293 to 393 K. The dielectric constant and energy density were increased with Bi2O3 NF content. This implies that we can utilize the prepared NC for the fabrication of energy storage devices. The influences of the Bi2O3 NF doping level on the transmittance, band gap, Urbach energy, refractive index, conductivity, and other physical parameters were discussed. The modifications induced in the dielectric/optical parameters make the prepared NC films the best candidates for applications requiring a high refractive index, such as optical coatings and integrated photonic devices.

Automated segmentation of cell organelles in volume electron microscopy using deep learning.

Recent advances in computing power triggered the use of artificial intelligence in image analysis in life sciences. To train these algorithms, a large enough set of certified labeled data is required. The trained neural network is then capable of producing accurate instance segmentation results that will then need to be re-assembled into the original dataset: the entire process requires substantial expertise and time to achieve quantifiable results. To speed-up the process, from cell organelle detection to quantification across electron microscopy modalities, we propose a deep-learning based approach for fast automatic outline segmentation (FAMOUS), that involves organelle detection combined with image morphology, and 3D meshing to automatically segment, visualize and quantify cell organelles within volume electron microscopy datasets. From start to finish, FAMOUS provides full segmentation results within a week on previously unseen datasets. FAMOUS was showcased on a HeLa cell dataset acquired using a focused ion beam scanning electron microscope, and on yeast cells acquired by transmission electron tomography. RESEARCH HIGHLIGHTS: Introducing a rapid, multimodal machine-learning workflow for the automatic segmentation of 3D cell organelles. Successfully applied to a variety of volume electron microscopy datasets and cell lines. Outperforming manual segmentation methods in time and accuracy. Enabling high-throughput quantitative cell biology.

Wound Dressing Based on Silver Nanoparticle Embedded Wool Keratin Electrospun Nanofibers Deposited on Cotton Fabric: Preparation, Characterization, Antimicrobial Activity, and Cytocompatibility.

Wool keratin (WK) protein is attractive for wound dressing and biomedical applications due to its excellent biodegradability, cytocompatibility, and wound-healing properties. In this work, WK-based wound dressings were prepared by depositing WK/poly(vinyl alcohol) (PVA) and silver nanoparticle (Ag NP)-embedded WK/PVA composite nanofibrous membranes on cotton fabrics by electrospinning. Ag NPs were biosynthesized by reduction and stabilization with sodium alginate. The formed Ag NPs were characterized by ultraviolet-visible and Fourier transform infrared (FTIR) spectroscopy, and their size was determined by transmission electron microscopy and image analysis. The formed Ag NPs were spherical and had an average diameter of 9.95 nm. The produced Ag NP-embedded WK/PVA composite nanofiber-deposited cotton fabric surface was characterized by FTIR and dynamic contact angle measurements, and the nanofiber morphologies were characterized by scanning electron microscopy. The average diameter of the nanofibers formed by 0.1% Ag NP-embedded WK/PVA solution was 146.7 nm. The antibacterial activity of the surface of cotton fabrics coated with electrospun composite nanofibers was evaluated against the two most common wound-causing pathogens, Staphylococcus aureus and Pseudomonas aeruginosa. The cotton fabric coated with 0.1% Ag NP-embedded WK/PVA nanofibers showed very good antibacterial activity against both pathogens, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay results showed good cytocompatibility against L-929 mouse fibroblast cells. However, the increase in Ag NP content in the nanofibers to 0.2% negatively affected the cell viability due to the high release rate of Ag ions. The results achieved show that the developed wound dressing has good potential for wound healing applications.

Tooth acellular extrinsic fibre cementum incremental lines in humans are formed by parallel branched Sharpey’s fibres and not by its mineral phase

In humans, the growth pattern of the acellular extrinsic fibre cementum (AEFC) has been useful to estimate the age-at-death. However, the structural organization behind such a pattern remains poorly understood. In this study tooth cementum from seven individuals from a Mexican modern skeletal series were analyzed with the aim of unveiling the AEFC collagenous and mineral structure using multimodal imaging approaches. The organization of collagen fibres was first determined using: light microscopy, transmission electron microscopy (TEM), electron tomography, and plasma FIB scanning electron microscopy (PFIB-SEM) tomography. The mineral properties were then investigated using: synchrotron small-angle X-ray scattering (SAXS) for T-parameter (correlation length between mineral particles); synchrotron X-ray diffraction (XRD) for L-parameter (mineral crystalline domain size estimation), alignment parameter (crystals preferred orientation) and lattice parameters a and c; as well as synchrotron X-ray fluorescence for spatial distribution of calcium, phosphorus and zinc. Results show that Sharpey’s fibres branched out fibres that cover and uncover other collagen bundles forming aligned arched structures that are joined by these same fibres but in a parallel fashion. The parallel fibres are not set as a continuum on the same plane and when they are superimposed project the AEFC incremental lines due to the collagen birefringence. The orientation of the apatite crystallites is subject to the arrangement of the collagen fibres, and the obtained parameter values along with the elemental distribution maps, revealed this mineral tissue as relatively homogeneous. Therefore, no intrinsic characteristics of the mineral phase could be associated with the alternating AEFC incremental pattern.

Strain-modulated Mn-rich layered oxide enables highly stable potassium-ion batteries

Mn-rich layered oxides show great promise as cathode materials for potassium-ion batteries due to their high capacity and cost-effectiveness. However, internal structural strain and irreversible phase transitions caused by Jahn-Teller distortion affect their cycling stability. Here, we present an efficient strategy to concurrently modulate the internal strain and suppress the irreversible phase transition in Mn-rich cathodes by incorporating amounts of zinc (Zn) ions into the transition metal layers. The substituted Zn serves to regulate local chemistry, thereby mitigating octahedral distortion in Mn-O bonds, relieving the strain between layers, and reducing the occurrence of P3-O3 phase transition under high voltages. These findings are supported by EXAFS, in situ X-ray diffraction, advanced transmission electron microscopy, electron tomography, and DFT simulations. The low-strain K0.5Mn0.8Co0.1Zn0.1O2 electrode exhibits a high reversible capacity retention about ∼90 % (105 mAh g−1 at 100 mA g−1), exceptional rate performance in the voltage of 1.5 ∼ 3.9 V, and a substantial capacity retention after 500 cycles. This strain-relieved approach broadens the scope of lattice engineering by addressing internal strain concerns and mitigating the strain associated with potassium (de)intercalation, thereby having potential to advance the development of stable cathodes for PIBs.

Flame‐assisted ultrafast synthesis of functionalized carbon nanosheets for high‐performance sodium storage

AbstractThe unique structural features of hard carbon (HC) make it a promising anode candidate for sodium‐ion batteries (SIB). However, traditional methods of preparing HC require special equipment, long reaction times, and large energy consumption, resulting in low throughputs and efficiency. In our contribution, a novel synthesis method is proposed, involving the formation of HC nanosheets (NS‐CNs) within minutes by creating an anoxic environment through flame combustion and further introducing sulfur and nitrogen sources to achieve heteroatom doping. The effect of heterogeneous element doping on the microstructure of HC is quantitatively analyzed by high‐resolution transmission electron microscopy and image processing technology. Combined with density functional theory calculation, it is verified that the functionalized HC exhibits stronger Na+ adsorption ability, electron gain ability, and Na+ migration ability. As a result, NS‐CNs as SIB anodes provide an ultrahigh reversible capacity of 542.7 mAh g−1 at 0.1 A g−1, and excellent rate performance with a reversible capacity of 236.4 mAh g−1 at 2 A g−1 after 1200 cycles. Furthermore, full cell assembled with NS‐CNs as the can present 230 mAh g−1 at 0.5 A g−1 after 150 cycles. Finally, in/ex situ techniques confirm that the excellent sodium storage properties of NS‐CNs are due to the construction of abundant active sites based on the novel synthesis method for realizing the reversible adsorption of Na+. This work provides a novel strategy to develop novel carbons and gives deep insights for the further investigation of facile preparation methods to develop high‐performance carbon anodes for alkali‐ion batteries.

GoldDigger and Checkers, computational developments in cryo-scanning transmission electron tomography to improve the quality of reconstructed volumes.

In this work, we present a pair of tools to improve the fiducial tracking and reconstruction quality of cryo-scanning transmission electron tomography (STET) datasets. We then demonstrate the effectiveness of these two tools on experimental cryo-STET data. The first tool, GoldDigger, improves the tracking of fiducials in cryo-STET by accommodating the changed appearance of highly defocussed fiducial markers. Since defocus effects are much stronger in scanning transmission electron microscopy than in conventional transmission electron microscopy, existing alignment tools do not perform well without manual intervention. The second tool, Checkers, combines image inpainting and unsupervised deep learning for denoising tomograms. Existing tools for denoising cryo-tomography often rely on paired noisy image frames, which are unavailable in cryo-STET datasets, necessitating a new approach. Finally, we make the two software tools freely available for the cryo-STET community.

Synthesis of nanocomposites of montmorillonite with carbon nanotubes as a potential material for water purification

The main goal of this research is to create nanocomposites based on unmodified and iron-modified (FeNP) montmorillonite (Mt) and carbon nanotubes (CNT) synthesized using the chemical vapor deposition method. The target area for the application of these materials is the creation of water treatment systems. This paper compares the efficiency of the CNT synthesis process on Mt before and after modification with FeNP of different concentrations and provides the characterization of the CNT microstructure and structure using different methods, such as scanning electron microscopy, high-resolution transmission electron imaging, and Raman spectroscopy. For initial verification of properties important for water purification, Mt+CNT and Mt+FeNP+CNT nanocomposites on a carbon nonwoven fabric (CF) are tested in this work. Incubation of the above-mentioned samples in a water–oil mixture reveals complex adsorption dynamics. The CF+Mt+FeNP+CNT sample shows a very good oil adsorption capacity due to its superhydrophobic and oleophilic properties.

An introduction to scanning transmission electron microscopy for the study of protozoans.

Since its inception in the 1930s, transmission electron microscopy (TEM) has been a powerful method to explore the cellular structure of parasites. TEM usually requires samples of <100 nm thick and with protozoans being larger than 1 μm, their study requires resin embedding and ultrathin sectioning. During the past decade, several new methods have been developed to improve, facilitate, and speed up the structural characterisation of biological samples, offering new imaging modalities for the study of protozoans. In particular, scanning transmission electron microscopy (STEM) can be used to observe sample sections as thick as 1 μm thus becoming an alternative to conventional TEM. STEM can also be performed under cryogenic conditions in combination with cryo-electron tomography providing access to the study of thicker samples in their native hydrated states in 3D. This method, called cryo-scanning transmission electron tomography (cryo-STET), was first developed in 2014. This review presents the basic concepts and benefits of STEM methods and provides examples to illustrate the potential for new insights into the structure and ultrastructure of protozoans.

ZnMn2O4 spinel nanocrystals-decorated multi-walled carbon nanotubes for oxygen reduction: Experimental and theoretical studies on the strong coupling facilitated four-electron selectivity

In this work, we have successfully formulated a single-step hydrothermal synthesis of mixed transition metal oxide nanocrystals (ZnMn2O4) strongly coupled on carbon support (MWCNTs) that acts as an efficient electrocatalyst for the electrochemical oxygen reduction reaction (ORR). The results depict that the composite, MWCNTs@ZnMn2O4 shows better electrocatalytic activity as compared to the activity exhibited by its individual components MWCNTs and ZnMn2O4 combined, attributed to the strong coupling between the two components. Transmission electron images of the composite depict that ZnMn2O4 nanocrystals have been successfully integrated on MWCNTs resulting in high conductivity and activity. The composite, MWCNTs@ZnMn2O4 exhibits an onset potential of 0.9 V vs. RHE, 99 % four-electron ORR selectivity, and high stability. The high selectivity and stability are assigned due to the strong coupling of ZnMn2O4 to MWCNTs through Zn/Mn–O–C bonds which are proved from DFT studies. The presence of Zn/Mn–O–C bonds is verified from X-ray photoelectron spectroscopy.

Improving Stability of CO2 Electroreduction by Incorporating Ag NPs in N-Doped Ordered Mesoporous Carbon Structures.

The electroreduction of carbon dioxide (eCO2RR) to CO using Ag nanoparticles as an electrocatalyst is promising as an industrial carbon capture and utilization (CCU) technique to mitigate CO2 emissions. Nevertheless, the long-term stability of these Ag nanoparticles has been insufficient despite initial high Faradaic efficiencies and/or partial current densities. To improve the stability, we evaluated an up-scalable and easily tunable synthesis route to deposit low-weight percentages of Ag nanoparticles (NPs) on and into the framework of a nitrogen-doped ordered mesoporous carbon (NOMC) structure. By exploiting this so-called nanoparticle confinement strategy, the nanoparticle mobility under operation is strongly reduced. As a result, particle detachment and agglomeration, two of the most pronounced electrocatalytic degradation mechanisms, are (partially) blocked and catalyst durability is improved. Several synthesis parameters, such as the anchoring agent, the weight percentage of Ag NPs, and the type of carbonaceous support material, were modified in a controlled manner to evaluate their respective impact on the overall electrochemical performance, with a strong emphasis on operational stability. The resulting powders were evaluated through electrochemical and physicochemical characterization methods, including X-ray diffraction (XRD), N2-physisorption, Inductively coupled plasma mass spectrometry (ICP-MS), scanning electron microscopy (SEM), SEM-energy-dispersive X-ray spectroscopy (SEM-EDS), high-angle annular dark field scanning transmission electron microscopy (HAADF-STEM), STEM-EDS, electron tomography, and X-ray photoelectron spectroscopy (XPS). The optimized Ag/soft-NOMC catalysts showed both a promising selectivity (∼80%) and stability compared with commercial Ag NPs while decreasing the loading of the transition metal by more than 50%. The stability of both the 5 and 10 wt % Ag/soft-NOMC catalysts showed considerable improvements by anchoring the Ag NPs on and into a NOMC framework, resulting in a 267% improvement in CO selectivity after 72 h (despite initial losses) compared to commercial Ag NPs. These results demonstrate the promising strategy of anchoring Ag NPs to improve the CO selectivity during prolonged experiments due to the reduced mobility of the Ag NPs and thus enhanced stability.

Self-Assembly of Nanofilaments in Cyanobacteria for Protein Co-localization.

Cyanobacteria offer great potential as alternative biotechnological hosts due to their photoautotrophic capacities. However, in comparison to established heterotrophic hosts, several key aspects, such as product titers, are still lagging behind. Nanobiotechnology is an emerging field with great potential to improve existing hosts, but so far, it has barely been explored in microbial photosynthetic systems. Here, we report the establishment of large proteinaceous nanofilaments in the unicellular model cyanobacterium Synechocystis sp. PCC 6803 and the fast-growing cyanobacterial strain Synechococcus elongatus UTEX 2973. Transmission electron microscopy and electron tomography demonstrated that expression of pduA*, encoding a modified bacterial microcompartment shell protein, led to the generation of bundles of longitudinally aligned nanofilaments in S. elongatus UTEX 2973 and shorter filamentous structures in Synechocystis sp. PCC 6803. Comparative proteomics showed that PduA* was at least 50 times more abundant than the second most abundant protein in the cell and that nanofilament assembly had only a minor impact on cellular metabolism. Finally, as a proof-of-concept for co-localization with the filaments, we targeted a fluorescent reporter protein, mCitrine, to PduA* by fusion with an encapsulation peptide that natively interacts with PduA. The establishment of nanofilaments in cyanobacterial cells is an important step toward cellular organization of heterologous pathways and the establishment of cyanobacteria as next-generation hosts.

Dislocations in naturally deformed olivine: Example of a mylonitic peridotite

We have investigated the microstructure of naturally deformed olivine (chemically equilibrated at 1000 °C) by conventional transmission electron microscopy and electron tomography. The peridotite specimen, from Oman ophiolite, has a mylonitic microstructure with remnant, strongly deformed, millimetric porphyroclasts co-existing with small newly formed olivine grains generated by dynamic recrystallization. Imaging by transmission electron microscopy reveals that both newly formed grains and porphyroclasts display [100] and [001] dislocations activity. Subgrain boundaries are composed of either [100] or [001] dislocations. The characterization of this natural sample also permits to identify sporadic [100] dislocation loops, rare [010] dislocation, infrequent melt, and intragranular bubbles or along subgrain boundaries. Electron tomography permits to identify several glide planes, which are similar to previous observations acquired on experimentally deformed polycrystalline olivine, more importantly electron tomography also permits to evidence combination of glide, climb and mixed climb (dislocation moving in an intermediate plane between the plane of glide plane and the plane of pure climb). Our study further infers the diversity of linear defects responsible for plastic deformation of olivine at lithospheric conditions.

Three-dimensional reconstruction of interstitial extracellular vesicles in human liver as determined by electron tomography.

Extracellular vesicles (EVs) are lipid bilayer nanoparticles involved in cell-cell communication that are released into the extracellular space by all cell types. The cargo of EVs includes proteins, lipids, nucleic acids, and metabolites reflecting their cell of origin. EVs have recently been isolated directly from solid tissues, and this may provide insights into how EVs mediate communication between cells in vivo. Even though EVs have been isolated from tissues, their point of origin when they are in the interstitial space has been uncertain. In this study, we performed three-dimensional (3D) reconstruction using transmission electron tomography of metastatic and normal liver tissues with a focus on the presence of EVs in the interstitium. After chemical fixation of the samples and subsequent embedding of tissue pieces in resin, ultrathin slices (300nm) were cut and imaged on a 120ekV transmission electron microscopy as a tilt series (a series of subsequent images tilted at different angles). These were then computationally illustrated in a 3D manner to reconstruct the imaged tissue volume. We identified the cells delimiting the interstitial space in both types of tissues, and small distinct spherical structures with a diameter of 30-200nm were identified between the cells. These round structures appeared to be more abundant in metastatic tissue compared to normal tissue. We suggest that the observed spherical structures in the interstitium of the metastatic and non-metastatic liver represent EVs. This work thus provides the first 3D visualization of EVs in human tissue.

Editors’ Choice—Structural, Electrical, and Luminescent Properties of Orthorhombic κ-Ga2O3 Grown by Epitaxial Lateral Overgrowth

The properties of orthorhombic κ-Ga2O3 films grown by Epitaxial Lateral Overgrowth (ELOG) were studied by Scanning Transmission Electron Microscopy (STEM), X-ray diffraction, capacitance-voltage profiling, Microcathodoluminescence (MCL) spectroscopy and imaging. ELOG mask was formed by deposition of SiO2 stripes on TiO2 buffer prepared on basal plane sapphire, with the stripes going along the [110] direction of sapphire. κ-Ga2O3 ELOG growth was performed using Halide Vapor Phase Epitaxy (HVPE), with ELOG wing of the structure formed by lateral overgrowth over the 20 μm-wide SiO2 stripes, while growth in between the stripes proceeded initially by vertical growth in the 5-μm-wide windows. TEM analysis showed that the material in the windows comprised 120o rotational nanodomains typical of κ-Ga2O3, while, in the wing regions, the material was single-domain monocrystalline. The films were conducting, with the net donor density close to 1013 cm−3. The data suggested the material in the windows have much higher resistance than in the wings. MCL spectra and imaging revealed much higher density of nonradiative recombination centers in the windows than in the wings.

Morphodynamics of non-canonical autophagic structures in Neurospora crassa.

Neurospora is a quintessential tip-growing organism, which is well known for packaging and longitudinal transport of tip-building blocks. Thus far, however, little attention has been paid to the co-essential process of reclamation, that is-taking apart of upstream, older structural elements, otherwise known as "autophagy". We are not yet prepared to set out the chemistry of that elaborate process, but its morphological start alone is worthy of attention. Carbon starvation triggers significant autophagic changes, beginning with prolific vacuolation along the plasma membrane, and eventual filling of 70% (or more) of cytoplasmic volume. Additionally, the Neurospora plasma membrane elaborates a variety of phagophores which themselves often look lytic. These have either dual enclosing membranes, like the familiar autophagosomes, can be doubled and have four wrapping membranes, or can be compounded with multiple membrane layers. These reclamation processes must be accommodated by the mechanism of tip growth.

Advances in imaging, scattering, spectroscopy, and machine learning-aided approaches for multiscale characterization of cementitious systems

Recent progress in methods used in the multiscale characterization of cementitious systems is reviewed, focusing on advances in imaging, scattering, and spectroscopy. The review includes relevant applications and developments in machine learning and other data analytics approaches to enhance characterization. Developments in imaging using light and electron microscopy as well as x-ray (i.e., from synchrotron) methods are summarized. Updates include scanning electron microscopy (SEM), transmission electron microscopy (TEM), tomography, and holography. A critical overview of spectroscopy (e.g., MAS NMR, Raman) and scattering (e.g., neutron, x-ray, synchrotron x-ray) methods is provided, and the intersection of these with imaging is developed (e.g., Raman imaging). Additionally, the paper summarizes recent developments in and implementations of state-of-the-art machine-learning algorithms and data analytics methods for automated, systematic, and/or quantitative analyses of image data sets.The review considers but is not limited to the application of these methods for investigating the hydration and microstructure development of cement phases, low-carbon-footprint cements (e.g., limestone calcined clay cements, LC3), environmental interactions (e.g., ASR) and model systems. Thus, the present work provides a critical presentation of advances in characterization methods that link together the composition and multiscale structure of cementitious materials.

Development of Hydroxyapatite Coatings for Orthopaedic Implants from Colloidal Solutions: Part 2-Detailed Characterisation of the Coatings and Their Growth Mechanism.

This study is the second part of a two-part study whereby supersaturated solutions of calcium and phosphate ions generate well-defined hydroxyapatite coatings for orthopaedic implants. An 'ideal' process solution is selected from Part 1, and the detailed characterisation of films produced from this solution is undertaken here in Part 2. Analysis is presented on the hydroxyapatite produced, in both powder form and as a film upon titanium substrates representative of orthopaedic implants. From thermal analysis data, it is shown that there is bound and interstitial water present in the hydroxyapatite. Nuclear magnetic resonance data allow for the distinction between an amorphous and a crystalline component of the material. As hydroxyapatite coatings are generated, their growth mechanism is tracked across repeated process runs. A clear understanding of the growth mechanism is achieved though crystallinity and electron imaging data. Transmission electron imaging data support the proposed crystal growth and deposition mechanism. All of the data conclude that this process has a clear propensity to grow the hydroxyapatite phase of octacalcium phosphate. The investigation of the hydroxyapatite coating and its growth mechanism establish that a stable and reproducible process window has been identified. Precise control is achieved, leading to the successful formation of the desired hydroxyapatite films.

Ptychographic X-ray computed tomography of porous membranes with nanoscale resolution

New visualization methods can be utilized to resolve structures at resolutions that were previously unachievable. 3D images of porous materials with high resolution have been so far obtained using transmission electron tomography or focused ion beam coupled with scanning electron microscopy. For these methods, ultra-vacuum is required, and only a small volume of the sample is visualized. Here, we demonstrate the application of ptychographic X-ray computed tomography for the visualization of soft matter with a resolution of 26 nm over large fields of view. Thanks to the high-penetration depth of the X-ray beam, we visualize the 3D complex porous structure of polyetherimide hollow fibers in a non-destructive manner and obtain quantitative information about pore size distribution and pore network interconnectivity across the whole membrane wall. The non-destructive nature of this method, coupled with its ability to image samples without requiring modification or a high vacuum environment, makes it valuable in the fields of porous- and nano-material sciences enabling imaging under different environmental conditions.

Isolation of Nanocellulose by Enzymatic Hydrolysis of Bleached Musa textilis (Abaca) Pulp

Nanocellulose is commonly isolated from cellulosic materials by chemical methods using strong acids. In this study, the enzymatic method was explored to isolate nanocellulose from commercial bleached S 2 grade abaca pulp. It was first disintegrated for 10 min and was subjected to enzyme hydrolysis while incubated with Bacillus sp. cellulase for 72 h at 50 °C and 120 revolutions/min. A clear liquid material was obtained after enzymatic hydrolysis after a series of centrifugation and ultrasonication. Results showed that the isolated nanocellulose had an average particle size of 375.9 nm ± 2.9 with a polydisperse index of 0.404 ± 0.059. Transmission electron and atomic force images showed that nanocellulose was longitudinal in size and highly aggregated and agglomerated. Through FTIR analysis, crystallinity indexes (i.e. lateral order index, total crystallinity index, and hydrogen bond intensity) of bleached abaca pulp and the isolated nanocellulose were compared. Results showed that the enzymatic hydrolysis of bleached abaca pulp resulted in higher cellulose crystallinity. Overall, nanocellulose can be isolated using biological methods using Bacillus sp. cellulase. These results could be used as a baseline to isolate smaller particle sizes, highly monodisperse, and stable nanocellulose that could be further applied in packaging, papermaking, cosmetics, medicine, and numerous other applications.