Three-dimensional Image Analysis Research Articles

Wavelength Dependence of Ultrahigh-Resolution Optical Coherence Tomography Using Supercontinuum for Biomedical Imaging

Optical coherence tomography (OCT) is a noninvasive cross-sectional imaging technique with micrometer resolution. The theoretical axial resolution is determined by the center wavelength and bandwidth of the light source, and the wider the bandwidth, the higher the axial resolution. The characteristics of OCT imaging depend on the optical wavelength used. In this paper, we investigated the wavelength dependence of ultrahigh-resolution (UHR) OCT using a supercontinuum for biomedical imaging. Wideband, high-power, low-noise supercontinua (SC) were generated at λ = 0.8, 1.1, 1.3, and 1.7 μ m based on ultrashort pulses and nonlinear fibers. The wavelength dependence of OCT imaging was examined quantitatively using biological phantoms. Ultrahigh-resolution imaging of a rat lung was demonstrated with λ = 0.8–1.0 μ m UHR-OCT. The variation of alveolar volume was estimated using three-dimensional image analysis. Finally, UHR-spectral domain-OCT and optical coherence microscopy at 1.7 μ m were developed, and high-resolution and high-penetration imaging of turbid tissue, especially mouse brain, was demonstrated.

Vitrifying immature equine oocytes impairs their ability to correctly align the chromosomes on the MII spindle.

Vitrified-warmed immature equine oocytes are able to complete the first meiotic division, but their subsequent developmental competence is compromised. Therefore, the present study investigated the effects of vitrifying immature horse oocytes on the chromosome and spindle configuration after IVM. Cumulus-oocytes complexes (COCs) were collected and divided into two groups based on mare age (young ≤14 years; old ≥16 years). COCs were then either directly matured invitro or vitrified and warmed before IVM. Spindle morphology and chromosome alignment within MII stage oocytes were assessed using immunofluorescent staining, confocal microscopy and three-dimensional image analysis. Vitrification reduced the ability of oocytes to reach MII and resulted in ultrastructural changes to the meiotic spindle, including shortening of its long axis, and an increased incidence of chromosomes failing to align properly at the metaphase plate. We hypothesise that aberrant chromosome alignment is an important contributor to the reduced developmental competence of vitrified equine oocytes. Contrary to expectation, oocytes from young mares were more severely affected than oocytes from older mares; we propose that the reduced effect of vitrification on oocytes from older mares is related to pre-existing compromise of spindle assembly checkpoint control mechanisms in these mares.

Utility of virtual three-dimensional image analysis for laparoscopic gastrectomy conducted by trainee surgeons.

Purpose The aim of this study was to investigate the utility the three-dimensional (3D) imaging for laparoscopic gastrectomy performed by trainee surgeons. Methods 3D-reconstruction was performed using multi-detector computed tomography (MDCT) and SYNAPSE VINCENT software. Trainee surgeons made 3D-imaging and checked the anatomical structure. Thirty-three patients who underwent laparoscopic gastrectomy (LG) for gastric cancer were examined. Trainees performed 19 LG, while specialists performed 14 LG. The vascular pattern and the surgical outcomes were evaluated. Result 3D imaging depicted the correct positional relationship between the gastric vasculatures and the organs. Regarding vascular pattern detected by 3D imaging, the origins of the infrapyloric artery were the right gastroepiploic artery in 12 cases (36%), the gastroduodenal artery in eight cases (24%), the bifurcation of the right gastroepiploic artery and gastroduodenal artery in seven cases (21%), and not detected in one case (3%). The types of confluence of the infrapyloric vein were the right gastroepiploic vein in 16 cases (48%), the anterior superior pancreatoduodenal vein in 10 cases (30%), and not detected in seven cases (21%). Surgical outcomes were not different between trainee group using intraoperative 3D image with the specialist in instruction group without the intraoperative 3D image. Conclusions Preoperative 3D imaging might contribute to successful and safe LG by trainee surgeons. J. Med. Invest. 66 : 280-284, August, 2019.

Three-dimensional volumetric magnetic resonance imaging (MRI) analysis of the soft palate and nasopharynx in brachycephalic and non-brachycephalic dog breeds.

The purpose of this study was to use magnetic resonance imaging (MRI) to compare the volumes and three-dimensional configurations of the soft palate and nasopharynx in non-brachycephalic and brachycephalic dogs with different body weights, and infer which factors influence nasopharyngeal volume. This was a retrospective observational study. The brain MRI medical records of all dogs referred to the Veterinary Medical Center, Chungbuk National University, between 2013 and 2016, for evaluation of intracranial disease were reviewed. There was a significant difference in the two-dimensional parameters including soft palate length/skull length ratio (P<0.01) and maximum soft palate thickness (P<0.01), and three-dimensional parameters which included soft palate volume (P<0.01), nasopharyngeal volume (P<0.01), soft palate/total upper airway volume ratio (P<0.01), and nasopharyngeal volume/total upper airway volume ratio (P<0.01), between brachycephalic and non-brachycephalic dog breeds. Nasopharyngeal volume correlated positively with the maximum soft palate thickness and body weight in all breeds. The three-dimensional morphologic grades of soft palate were significantly different between the two groups. In brachycephalic breeds, Grade 3 was observed in 33% of cases but was absent in non-brachycephalic breeds, where Grade 1 was present in 85% of the cases. We can conclude that three-dimensional morphology and upper airway volume are significantly different between brachycephalic and non-brachycephalic breeds, and body weight and maximum soft palate thickness are the key factors associated with a decreased nasopharyngeal volume.

X-ray microtomography to evaluate the efficacy of paraffin wax coating for soil bulk density evaluation

The paraffin-coated method is a well-used approach to measure the soil bulk density (BD). BD is a physical property of great importance for studies of soil quality and health. Therefore, representative measurements of this property are highly valued. Resin and paraffin wax are utilized to coat soil samples; however, if these materials ingress into the sample it could affect the representativeness of BD evaluation. The advance in three-dimensional (3D) image analysis techniques such as X-ray microtomography (μCT) offers a great opportunity to visualize and quantify the possible penetration of paraffin wax into clod samples. In this paper we investigated porous system morphological properties of soil samples coated with paraffin wax. The morphological properties of the pores filled with paraffin wax inside the samples were also studied. We observed qualitatively that samples with large pores close to their borders were more susceptible to the penetration of paraffin wax. Samples with pores >10 mm3 had the highest amount of paraffin wax into them. Triaxial shaped and complexly pores also offered less resistance to the ingress of paraffin wax. Positive relations between the amount of paraffin wax inside the samples and the volume of pores measured, pore tortuosity and degree of anisotropy were found. Conversely, the soil pore connectivity was not correlated with the penetration of paraffin wax into the samples, at least for the region of interest (≈27.3 cm3) studied. Finally, an analysis of the impact of paraffin wax ingress inside the samples in measured BD showed increments of ≈0.09 and ≈0.11 g cm−3 in this property when the paraffin wax penetrates into the large pores.

Dynamic In Situ Three-Dimensional Imaging and Digital Volume Correlation Analysis to Quantify Strain Localization and Fracture Coalescence in Sandstone

Advances in triaxial compression deformation apparatus design, dynamic X-ray microtomography imaging, data analysis techniques, and digital volume correlation analysis provide unparalleled access to the in situ four-dimensional distribution of developing strain within rocks. To demonstrate the power of these new techniques and acquire detailed information about the micromechanics of damage evolution, deformation, and failure of porous rocks, we deformed 3-cm-scale cylindrical specimens of low-porosity Fontainebleau sandstone in an X-ray-transparent triaxial compression apparatus, and repeatedly recorded three-dimensional tomograms of the specimens as the differential stress was increased until macroscopic failure occurred. Experiments were performed at room temperature with confining pressure in the range of 10–20 MPa. Distinct grayscale subsets, indicative of density, enabled segmentation of the three-dimensional tomograms into intact rock matrix, pore space, and fractures. Digital volume correlation analysis of pairs of tomograms provided time series of three-dimensional incremental strain tensor fields throughout the experiments. After the yield stress was reached, the samples deformed first by dilatant opening and propagation of microfractures, and then by shear sliding via grain rotation and strain localization along faults. For two samples, damage and dilatancy occurred by grain boundary opening and then a sudden collapse of the granular rock framework at failure. For the third sample, a fault nucleated near the yield point and propagated in the sample through the development of transgranular microfractures. The results confirm findings of previous experimental studies on the same rock and provide new detailed quantifications of: (1) the proportion of shear versus dilatant strain in the sample, (2) the amount of dilatancy due to microfracture opening versus pore opening when a fault develops, and (3) the role of grain boundaries and pore walls in pinning microfracture propagation and slowing down the rate of damage accumulation as failure is approached. Our study demonstrates how the combination of high-resolution in situ dynamic X-ray microtomography imaging and digital volume image correlation analysis can be used to provide additional information to unravel brittle failure processes in rocks under stress conditions relevant to the upper crust.

A quantitative morphological analysis of three-dimensional CT coxal bone images of contemporary Japanese using homologous models for sex and age estimation

Sexual dimorphisms and age-dependent morphological features of the human coxal bone were quantitatively analyzed using homologous models created from three-dimensional (3D) computed tomography images of the pelvis (male: 514 samples, female: 388 samples, age 16-100). Bilateral average coxal images of each sex and age decade were generated separately through principle component analyses (PCA). By measuring average point-to-point distances of 8472 corresponding points (average corresponding point differences [ACPDs]) between each homologous coxal image and the average images, the sex of more than 93% of the samples was correctly assigned. Some principal components (PCs) detected in PCA of the homologous models of the samples correlated fairly well with age and are affected by features of the curvature of the iliac crest, the arcuate line and the greater sciatic notch. Moreover, separate PCA using the average images of each age decade successfully detected the first PCs, which were strongly correlated with age. However, neither multiple regression analysis using PCs related to age nor comparison of ACPDs with the average images of each age decade could produce accurate results for age decade assignment of unknown (blind) samples. Therefore, more detailed analysis of age-dependent morphological features would be necessary for actual age estimation. In addition, some laterality or left and right shape difference of the coxal bone images was also elucidated, and was more significant in females. Analysis of 3D structures using homologous models and PCA appears to be a potential technique to detect subsistent morphological changes of bones.

P2.15-27 Lung Volume Change After Lobectomy Estimated by Three-Dimensional Image Analysis System

P2.15-27 Lung Volume Change After Lobectomy Estimated by Three-Dimensional Image Analysis System

Two-photon-excited, three-dimensional photoluminescence imaging and dislocation-line analysis of threading dislocations in 4H-SiC

This paper describes the three-dimensional imaging of threading edge dislocations (&amp;lt;a&amp;gt;TEDs), pure c threading screw dislocations (&amp;lt;c&amp;gt;TSDs), and c + a threading mixed dislocations (&amp;lt;c + a&amp;gt;TMDs) in 4H silicon carbide (4H-SiC) epilayers by two-photon-excited photoluminescence (2PPL), where inclinations of dislocations in epilayers with different offcut angles are investigated. A numerical model based on carrier diffusion explains the mechanism of the dark-contrast 2PPL imaging provided by band edge emission. More than 450 threading dislocations in 4H-SiC epilayers are visualized three-dimensionally and are classified into six &amp;lt;a&amp;gt;-type TEDs, two &amp;lt;c&amp;gt;-type TSDs, and twelve &amp;lt;c + a&amp;gt;-type TMDs according to the directions of extra half-planes and the chiral geometries (right- or left-handed screw). The 2PPL images reveal that all threading dislocation types incline in the step-flow direction, the extent of which is more prominent for a larger offcut angle. The &amp;lt;a&amp;gt;TEDs also incline in the directions of their extra half-planes. The inclinations of &amp;lt;c&amp;gt;TSDs perpendicular to the step-flow direction are influenced by chirality, while those of &amp;lt;c + a&amp;gt;TMDs are influenced by both the directions of the extra half-planes and the chirality. The origins of the dislocation inclinations are discussed taking into account the kinetic interactions between the dislocations and the advancing steps on the growing surface.

Three-dimensional microscopy and image analysis methodology for mapping and quantification of nuclear positions in tissues with approximate cylindrical geometry.

Organogenesis involves extensive and dynamic changes of tissue shape during development. It is associated with complex morphogenetic events that require enormous tissue plasticity and generate a large variety of transient three-dimensional geometries that are achieved by global tissue responses. Nevertheless, such global responses are driven by tight spatio-temporal regulation of the behaviours of individual cells composing these tissues. Therefore, the development of image analysis tools that allow for extraction of quantitative data concerning individual cell behaviours is central to study tissue morphogenesis. There are many image analysis tools available that permit extraction of cell parameters. Unfortunately, the majority are developed for tissues with relatively simple geometries such as flat epithelia. Problems arise when the tissue of interest assumes a more complex three-dimensional geometry. Here, we use the endothelium of the developing zebrafish dorsal aorta as an example of a tissue with cylindrical geometry and describe the image analysis routines developed to extract quantitative data on individual cells in such tissues, as well as the image acquisition and sample preparation methodology.This article is part of the Theo Murphy meeting issue 'Mechanics of development'.

Vertical Bone Augmentation Using Ring Technique with Three Different Materials in the Sheep Mandible Bone.

The purpose of this study was to examine the volumetric alterations and osseointegration properties in the augmented area of the ring technique using different types of bone graft material in sheep mandible bone. Three different materials (columnar forms, 7-mm diameter, 3-mm height) were stabilized using dental implants with a turned surface in the mandible bone of Finnish Dorset cross-bred sheep: group A, autogenous bone; group B, bovine bone; group C, biphasic bone substitute. Animals were euthanized after 5 weeks (N = 6). Three-dimensional image data by digital oral scanner were taken at the surgery and sacrifice, and the volume alteration of the material was calculated. The bone samples were fixed in formalin and dehydrated in ethanol. Resin-embedded samples were subjected to non-decalcified ground sectioning, and histologic and histomorphometric analysis (bone and material area alteration, bone-to-implant contact [BIC]) were done. In three-dimensional (3D) image analysis, group A showed a statistically higher percentage of remaining materials compared with groups B and C. The histologic observation showed no new bone formations around the implants in all groups, especially at the maxillary site of the implant in the augmented area. In histomorphometric analysis, group A showed a statistically higher percentage of bone area (BA) compared with groups B and C; however, in all groups, bone-to-implant contact (BIC) showed low values, and there were no statistical differences between groups. The results of this study suggested that the autogenous bone maintained bone volume around the dental implant using the ring technique, and the impact of surface properties was of some importance; osseointegration with the turned surface in the augmented area showed low BIC values in all groups.

Limitations of a 3-D Image Analysis-Based Particle Size Measuring System for Wood Particle Dimension Measurement

Against the background of inaccurately measured wood particle dimensions, applying the three-dimensional (3-D) image analysis–based particle size characterization system Partimac 3D XL in pre- liminary tests, metal platelets with various aspect ratios of the three main axes are employed to understand and explain the observed limitations of the measuring principle. It was found that particle width and thickness interact increasingly with a decreasing aspect ratio and, thus, the digital replica and subsequent determination of the dimensions become incorrect. This was ascribed to the random orientation of the particles during image acquisition, which cannot be overcome with a finite number of cameras in the system.

Mechanical Strength of the Proximal Femur After Arthroscopic Osteochondroplasty for Femoroacetabular Impingement: Finite Element Analysis and 3-Dimensional Image Analysis

To examine the influence of femoral neck resection on the mechanical strength of the proximal femur in actual surgery. Eighteen subjects who received arthroscopic cam resection for cam-type femoroacetabular impingement (FAI) were included. Finite element analyses (FEAs) were performed to calculate changes in simulative fracture load between pre- and postoperative femur models. The finite element femur models were constructed from computed tomographic images; thus, the models represented the shape of the original femur, including the bone resection site. Three-dimensional image analysis of the bone resection site was performed to identify morphometric factors that affect strength in the postoperative femur model. Four oblique sagittal planes running perpendicular to the femoral neck axis were used as reference planes to measure the bone resection site. At the transcervical reference plane, both the bone resection depth and the cross-sectional area at the resection site correlated strongly with postoperative changes in the simulated fracture load (R2= 0.6, P= .0001). However, only resection depth was significantly correlated with the simulated fracture load at the reference plane for the head-neck junction. The resected bone volume did not correlate with the postoperative changes in the simulated fracture load. The results of our FEA suggest that the bone resection depth measured at the head-neck junction and transcervical reference plane correlates with fracture risk after osteochondroplasty. By contrast, bone resection at more proximal areas did not have a significant effect on the postoperative femur model strength in our FEA. The total volume of resected bone was also not significantly correlated with postoperative changes in femur model strength. This biomechanical study using FEA suggest that there is a risk of femoral neck fracture after arthroscopic cam resection, particularly when the resected lesion is located distally.

Hounsfield unit of screw trajectory as a predictor of pedicle screw loosening after single level lumbar interbody fusion

ObjectThis study aims to clarify the clinical potential of Hounsfield unit (HU), measured on computed tomography (CT) images, as a predictor of pedicle screw (PS) loosening, compared to bone mineral density (BMD). MethodsA total of 206 screws in 52 patients (21 men and 31 women; mean age 68.2 years) were analyzed retrospectively. The screws were classified into two groups depending on their screw loosening status on 3-month follow-up CT (loosening screw group vs. non-loosening screw group). Preoperative HU of the trajectory was evaluated by superimposing preoperative and postoperative CT images using three-dimensional image analysis software. Age, sex, body mass index, screw size, BMD of lumbar, and HU of screw trajectory were analyzed in association with screw loosening. Multivariate logistic regression analysis was performed, and the thresholds for PS loosening risk factors were evaluated using a continuous numerical variable and receiver operating characteristic (ROC) curve analyses. The area under the curve (AUC) was used to determine the diagnostic performance, and values > 0.75 were considered to represent good performance. ResultsThe loosening screw group contained 24 screws (12%). Multivariate analysis revealed that the significant independent risk factors were not BMD but male sex [P = 0.028; odds ratio (OR) 2.852, 95% confidence interval (CI) 1.120–7.258] and HU of screw trajectory (P = 0.006; OR 0.989, 95% CI 0.980–0.997). ROC curve analysis demonstrated that the AUC for HU of screw trajectory for women was 0.880 (95% CI 0.798–0.961). The cutoff value was 153.5. AUC for men was 0.635 (95% CI 0.449–0.821), which was not considered to be a good performance. ConclusionsLow HU of screw trajectories was identified as a risk factor of PS loosening for women. For female patients with low HU, additional augmentation is recommended to prevent PS loosening.

Tomographic and multimodal scattering-type scanning near-field optical microscopy with peak force tapping mode

Scattering-type scanning near-field optical microscopy (s-SNOM) enables nanoscale spectroscopic imaging and has been instrumental for many nano-photonic discoveries and in situ studies. However, conventional s-SNOM techniques with atomic force microscopy tapping mode operation and lock-in detections do not provide direct tomographic information with explicit tip−sample distance. Here, we present a non-traditional s-SNOM technique, named peak force scattering-type scanning near-field optical microscopy (PF-SNOM), by combination of peak force tapping mode and time-gated light detection. PF-SNOM enables direct sectioning of vertical near-field signals from a sample surface for both three-dimensional near-field imaging and spectroscopic analysis. Tip-induced relaxation of surface phonon polaritons are revealed and modeled by considering tip damping. PF-SNOM also delivers a spatial resolution of 5 nm and can simultaneously measure mechanical and electrical properties together with optical near-field signals. PF-SNOM is expected to facilitate three-dimensional nanoscale near-field characterizations and correlative in situ investigations on light-induced mechanical and electrical effects.

Internal head morphology of minor workers and soldiers in the hyperdiverse ant genus Pheidole

In the hyperdiverse ant genus Pheidole Westwood, 1839, the worker caste evolved into two morphologically distinct subcastes: minor workers and soldiers. The evolution of soldiers, which are larger in size than minor workers and have disproportionately larger heads, are thought to be key to Phediole’s success. Although many studies have focused on external anatomy, little is known about their internal anatomy. We therefore used microCT imaging and quantitative three-dimensional image analysis to reconstruct the major glands of the head, the musculature, nervous system, and digestive organ of minor workers and soldiers of four Pheidole species. We expected these tissues to scale isometrically and to be proportionally larger in soldiers relative to the minor workers. Surprisingly, we found that the nervous system, cephalic gland, and digestive organ volume are absolutely and relatively smaller in soldiers, whereas muscle volume is absolutely and relatively larger, than in minor workers. This may reflect individual-level trade-offs, where muscles grow at the expense of all other cephalic organs. Alternatively, this relationship may reflect the specialization of internal anatomy in each subcaste to enhance division of labour at the colony level. Future studies should test these alternative hypotheses across a larger number of Pheidole species.

Genomic regions responsible for seminal and crown root lengths identified by 2D & 3D root system image analysis

BackgroundGenetic improvement of root system architecture is a promising approach for improved uptake of water and mineral nutrients distributed unevenly in the soil. To identify genomic regions associated with the length of different root types in rice, we quantified root system architecture in a set of 26 chromosome segment substitution lines derived from a cross between lowland indica rice, IR64, and upland tropical japonica rice, Kinandang Patong, (IK-CSSLs), using 2D & 3D root phenotyping platforms.ResultsLengths of seminal and crown roots in the IK-CSSLs grown under hydroponic conditions were measured by 2D image analysis (RootReader2D). Twelve CSSLs showed significantly longer seminal root length than the recurrent parent IR64. Of these, 8 CSSLs also exhibited longer total length of the three longest crown roots compared to IR64. Three-dimensional image analysis (RootReader3D) for these CSSLs grown in gellan gum revealed that only one CSSL, SL1003, showed significantly longer total root length than IR64. To characterize the root morphology of SL1003 under soil conditions, SL1003 was grown in Turface, a soil-like growth media, and roots were quantified using RootReader3D. SL1003 had larger total root length and increased total crown root length than did IR64, although its seminal root length was similar to that of IR64. The larger TRL in SL1003 may be due to increased crown root length.ConclusionsSL1003 carries an introgression from Kinandang Patong on the long arm of chromosome 1 in the genetic background of IR64. We conclude that this region harbors a QTL controlling crown root elongation.

Simulation and analysis of three-dimensional echo decorrelation imaging

A numerical model for pulse-echo ultrasound imaging [Mast, JASA 128:EL99-EL104, 2010] is extended to three-dimensional (3D) imaging of linear, weakly scattering continuum media by matrix array transducers. In this model, beam patterns for steered transmit and receive subapertures, calculated analytically under the Fresnel approximation, are combined with a 3D numerical tissue model to yield beamformed scan lines in a rectilinear or pyramidal configuration. Simulated scan lines are processed to yield both volumetric B-mode images and echo decorrelation images, comprising spatial maps of the normalized decorrelation between sequential pulse-echo image volumes. A method is demonstrated for construction and scan conversion of quantitative 3D echo decorrelation images such that, as for 2D echo decorrelation imaging [Hooi et al., JASA 137:585-597, 2015], the mapped echo decorrelation consistently estimates the local decoherence spectrum of the tissue reflectivity. Simulated 3D echo decorrelation images of random media are shown to accurately estimate known decoherence distributions that simulate heating effects in thermal ablation. Effects of factors including transducer frequency, scan configuration, and simulated lesion size on accuracy of decoherence estimates are tested. Applications to 3D monitoring of radiofrequency and microwave ablation are discussed.

Sequential 3-dimensional computed tomography analysis of implant position following total shoulder arthroplasty

Detection of postoperative component position and implant shift following total shoulder arthroplasty (TSA) can be challenging using routine imaging. The purpose of this study was to evaluate glenoid component position over time using 3-dimensional computed tomography (CT) analysis with minimum 2-year follow-up. Twenty patients underwent primary TSA with sequential CT scanning of the shoulder: a preoperative study, an immediate postoperative study within 2 weeks of surgery, and a postoperative study performed at minimum 2-year follow-up (CT3). Postoperative glenoid component position and central peg osteolysis were assessed across the immediate postoperative CT scan and CT3. Glenoids with evidence of component shift and/or grade 1 central peg osteolysis on CT3 were considered at risk of loosening. Of the patients, 7 (35%) showed evidence of glenoid components at risk of loosening on CT3, 6 with component shift (3 with increased inclination alone, 1 with increased retroversion alone, and 2 with both increased inclination and retroversion). Significantly more patients with glenoid component shift had grade 1 central peg osteolysis on CT3 compared with those without shift (83% vs 7%, P = .002). One clinical failure occurred, with the patient undergoing revision to reverse TSA for rotator cuff deficiency. Three-dimensional CT imaging analysis following TSA identified changes in glenoid component position over time, with inclination being the most common direction of shift and grade 1 central peg osteolysis commonly associated with shift. These findings raise concern for glenoids at risk of loosening, but further follow-up is needed to determine the long-term clinical impact of these findings.

Three dimensional typological studies using scanning electron microscopy for characterization of Termitomyces pellets obtained from submerged growth conditions

There are gaps in existing understanding of fungal pellet growth dynamics. We used scanning electron microscopy (SEM) for morphological characterization of the biomass organization of Termitomyces pellets for seven species: T. microcarpus (TMI1), T. albuminosus (TAL1, TAL2), T. striatus (TSTR), T. aurantiacus (TAUR), T. heimii (THE1, THE2), T. globulus (TGLO) and T. clypeatus (TCL1, TCL2, TCL3, TCL4, TCL5). We assessed the utility of SEM for morphological and structural characterization of Termitomyces spp. in three dimensional (3D) pellet form to identify ideal pellet morphology for industrial use. Typological classification of Termitomyces species was based on furrows, isotropy, total motifs and fractal dimensions. The pellets formed were entangled and exhibited highly compacted mycelial mass with microheterogeneity and microporosity. The mean density of furrows of Termitomyces species was between 10,000 and 11,300 cm/cm2, percentage isotropy was 30−80 and total motifs varied from 300 to 2500. TGLO exhibited the highest furrow mean density, 11243 cm/cm2, which indicated a compact, cerebroid structure with complex ridges and furrows, whereas TAL2 exhibited the lowest furrow density. TMI1a exhibited a high percentage isotropic value, 74.6, TSTR exhibited the lowest, 30.9. Total motif number also was used as a typological classification parameter. Fractal values were 2.64−2.78 for various submerged conditions of Termitomyces species. TAL1 exhibited the highest fractal dimension and TAL2 the lowest, which indicates the complexity of branching patterns. Three-dimensional SEM image analysis can provide insight into pellet micromorphology and is a powerful tool for exploring topographical details of pellets.