Evaluation of a 3D-Printed Cleft Palate Obturator Using a Low-Dose Cone Beam Computed Tomography Acquisition Protocol: A Proof-of-Concept Study.

Cone beam computed tomography (CBCT) technology is increasingly utilized in the head and neck region and is valuable in treatment planning for cleft palate patients, potentially enabling the creation of 3D-printed obturators to assist with feeding and speech. This technical report investigates the feasibility of using data from a 360-degree CBCT scan to accurately produce a cleft palate obturator and assesses whether a lower-dose 180-degree CBCT scan can achieve a comparable result. A simulated cleft palate was crafted on a dehydrated human skull, which was then scanned using both 360-degree and 180-degree CBCT scanning protocols. Two obturators were digitally designed based on the segmented images from each scan and subsequently 3D printed. Evaluation of the segmented images and 3D-printed obturators from both protocols demonstrated clear visualization of anatomical landmarks and identical scores across all parameters, suggesting that the 180-degree CBCT scan can produce an obturator of comparable quality to that of the 360-degree scan, with the added benefit of reduced radiation exposure.

Combining physics-based models with deep learning image synthesis and uncertainty in intraoperative cone-beam CT of the brain.

BACKGROUND: Image-guided neurosurgery requires high localization and registration accuracy to enable effective treatment and avoid complications. However, accurate neuronavigation based on preoperative magnetic resonance (MR) or computed tomography (CT) images is challenged by brain deformation occurring during the surgical intervention. PURPOSE: To facilitate intraoperative visualization of brain tissues and deformable registration with preoperative images, a 3D deep learning (DL) reconstruction framework (termed DL-Recon) was proposed for improved intraoperative cone-beam CT (CBCT) image quality. METHODS: The DL-Recon framework combines physics-based models with deep learning CT synthesis and leverages uncertainty information to promote robustness to unseen features. A 3D generative adversarial network (GAN) with a conditional loss function modulated by aleatoric uncertainty was developed for CBCT-to-CT synthesis. Epistemic uncertainty of the synthesis model was estimated via Monte Carlo (MC) dropout. Using spatially varying weights derived from epistemic uncertainty, the DL-Recon image combines the synthetic CT with an artifact-corrected filtered back-projection (FBP) reconstruction. In regions of high epistemic uncertainty, DL-Recon includes greater contribution from the FBP image. Twenty paired real CT and simulated CBCT images of the head were used for network training and validation, and experiments evaluated the performance of DL-Recon on CBCT images containing simulated and real brain lesions not present in the training data. Performance among learning- and physics-based methods was quantified in terms of structural similarity (SSIM) of the resulting image to diagnostic CT and Dice similarity metric (DSC) in lesion segmentation compared to ground truth. A pilot study was conducted involving seven subjects with CBCT images acquired during neurosurgery to assess the feasibility of DL-Recon in clinical data. RESULTS: CBCT images reconstructed via FBP with physics-based corrections exhibited the usual challenges to soft-tissue contrast resolution due to image non-uniformity, noise, and residual artifacts. GAN synthesis improved image uniformity and soft-tissue visibility but was subject to error in the shape and contrast of simulated lesions that were unseen in training. Incorporation of aleatoric uncertainty in synthesis loss improved estimation of epistemic uncertainty, with variable brain structures and unseen lesions exhibiting higher epistemic uncertainty. The DL-Recon approach mitigated synthesis errors while maintaining improvement in image quality, yielding 15%-22% increase in SSIM (image appearance compared to diagnostic CT) and up to 25% increase in DSC in lesion segmentation compared to FBP. Clear gains in visual image quality were also observed in real brain lesions and in clinical CBCT images. CONCLUSIONS: DL-Recon leveraged uncertainty estimation to combine the strengths of DL and physics-based reconstruction and demonstrated substantial improvements in the accuracy and quality of intraoperative CBCT. The improved soft-tissue contrast resolution could facilitate visualization of brain structures and support deformable registration with preoperative images, further extending the utility of intraoperative CBCT in image-guided neurosurgery.

Structural alterations in cell organelles induced by photodynamic treatment with chlorin p6 -histamine conjugate in human oral carcinoma cells probed by 3D fluorescence microscopy.

We have explored the intracellular cell organelle's structural alterations after photodynamic treatment with chlorin p6 -histamine conjugate (Cp6 -his) in human oral cancer cells. Herein, the cells were treated with Cp6 -his (10 µm) and counterstained with organelle-specific fluorescence probes to find the site of intracellular localization using confocal microscopy. For photodynamic therapy (PDT), the cells were exposed to ~30 kJ/m2 red light (660 ± 20 nm) to induce ~90% cytotoxicity. We used the three-dimensional (3D) image reconstruction approach to analyze the photodynamic damage to cell organelles. The result showed that Cp6 -his localized mainly in the endoplasmic reticulum (ER) and lysosomes but not in mitochondria and Golgi apparatus (GA). The 3D model revealed that in necrotic cells, PDT led to extensive fragmentation of ER and fragmentation and swelling of GA as well. Results suggest that the indirect damage to GA occurred due to loss of connection between ER and GA. Moreover, in damaged cells with no sign of necrosis, the perinuclear ER appeared condensed and surrounded by several small clumps at the peripheral region of the cell, and the GA was observed to form a single condensed structure. Since these structural changes were associated with apoptotic cell death, it is suggested that the necrotic and apoptotic death induced by PDT with Cp6 -his is determined by the severity of damage to ER and indirect damage to GA. The results suggest that the indirect damage to cell organelle apart from the sites of photosensitizer localization and the severity of damage at the organelle level contribute significantly to the mode of cell death in PDT.

An Innovative Concept of a 3D-Coded Aperture Imaging System Proposed for Early Breast Cancer Detection.

Coded Aperture (CA) imaging has recently been used in nuclear medicine, but still, there is no commercial SPECT imaging camera based on CA for cancer detection. The literature is rich in examples of using the CA for planar and thin 3D imaging. However, thick 3D reconstruction is still challenging for small lesion detection. This paper presents the results of mosaic modified uniformly redundant array (MURA) mask/antimask CA combined with a maximum-likelihood expectation-maximization (MLEM) algorithm. The MLEM is an iterative algorithm applied to a mosaic MURA CA mask/antimask for 3D anthropomorphic breast phantom reconstruction, slice by slice. The difference between the mask and the antimask suppresses the background noise to enhance the quality of reconstructed images. Furthermore, all reconstructed slices are stacked to form a 3D breast phantom image from single-projection data. The results of phantom reconstruction with 8 mm, 6 mm, 4 mm, and 3 mm lesions are presented. Moreover, the proposed SPECT imaging camera can reconstruct a 3D breast phantom from single-projection data of the patient's scanning. To assess the quality of lesions in the reconstructed images, the contrast-to-background ratio (CBR), the peak signal-to-noise ratio (PSNR) and mean square error (MSE) were measured.

A geometry-informed deep learning framework for ultra-sparse 3D tomographic image reconstruction.

Deep learning affords enormous opportunities to augment the armamentarium of biomedical imaging. However, the pure data-driven nature of deep learning models may limit the model generalizability and application scope. Here we establish a geometry-informed deep learning framework for ultra-sparse 3D tomographic image reconstruction. We introduce a novel mechanism for integrating geometric priors of the imaging system. We demonstrate that the seamless inclusion of known priors is essential to enhance the performance of 3D volumetric computed tomography imaging with ultra-sparse sampling. The study opens new avenues for data-driven biomedical imaging and promises to provide substantially improved imaging tools for various clinical imaging and image-guided interventions.

A Novel Task-Based reconstruction approach for digital breast tomosynthesis.

The reconstruction of a volumetric image from Digital Breast Tomosynthesis (DBT) measurements is an ill-posed inverse problem, for which existing iterative regularized approaches can provide a good solution. However, the clinical task is somehow omitted in the derivation of those techniques, although it plays a primary role in the radiologist diagnosis. In this work, we address this issue by introducing a novel variational formulation for DBT reconstruction, tailored for a specific clinical task, namely the detection of microcalcifications. Our method aims at simultaneously enhancing the detectability performance and enabling a high-quality restoration of the background breast tissues. Our contribution is threefold. First, we introduce an original task-based reconstruction framework through the proposition of a detectability function inspired from mathematical model observers. Second, we propose a novel total-variation regularizer where the gradient field accounts for the different morphological contents of the imaged breast. Third, we integrate the two developed measures into a cost function, minimized thanks to a new form of the Majorize Minimize Memory Gradient (3MG) algorithm. We conduct a numerical comparison of the convergence speed of the proposed method with those of standard convex optimization algorithms. Experimental results show the interest of our DBT reconstruction approach, qualitatively and quantitatively.

An alternative approach to tracing the volumic proliferation development of an entire tumor spheroid in 3D through a mini-Opto tomography platform.

Microscopy, which is listed among the major in-situ imaging applications, allows to derive information from a biological sample on the existing architectural structures of cells and tissues and their changes over time. Large biological samples such as tumor spheroids cannot be imaged within one field of view, regional imaging in different areas and subsequent stitching are required to attain the full picture. Microscopy is not typically used to produce full-size visualization of tumor spheroids measuring a few millimeters in size. In this study, we propose a 3D volume imaging technique for tracing the growth of an entire tumor spheroid measuring up to 10 mm using a miniaturized optical (mini-Opto) tomography platform. We performed a primary analysis of the 3D imaging for the MIA PaCa-2 pancreatic tumoroid employing its 2D images produced with the mini-Opto tomography from different angles ranging from -25 ° to +25 ° at six different three-day-apart time points of consecutive image acquisition. These 2D images were reconstructed by using a 3D image reconstruction algorithm that we developed based on the algebraic reconstruction technique (ART). We were able to reconstruct the 3D images of the tumoroid to achieve 800 × 800-pixel 50-layer images at resolutions of 5-25 µm. We also created its 3D visuals to understand more clearly how its volume changed and how it looked over weeks. The volume of the tumor was calculated to be 6.761 mm3 at the first imaging time point and 46.899 mm3 15 days after the first (at the sixth time point), which is 6.94 times larger in volume. The mini-Opto tomography can be considered more advantageous than commercial microscopy because it is portable, more cost-effective, and easier to use, and enables full-size visualization of biological samples measuring a few millimeters in size.

Reconstruction of 3D greyscale image for reservoir rock from a single image based on pattern dictionary.

Most methods that model 3D porous media from 2D images are based on binary images. In this paper, we propose a method for reconstructing 3D greyscale isotropic porous media images from a single image. Our proposed method incorporates a fast-sampling procedure to control the continuity and variability between adjoining reconstruction layers, a new similarity calculation method to obtain the most similar patterns from a pattern dictionary, and a central area simulation procedure to solve the block effect problem. The reconstruction results from application of our proposed method to a real reservoir 3D model obtained via computed tomography (CT) and a comparison with the original CT structure demonstrate that our proposed method can reproduce properties such as autocorrelation function, linear function, shape distribution, average shape factor, average pore radius size, average throat radius size, average pore volume, permeability and grey histogram. Further, the comparison results indicate that the statistical characteristics of the reconstructions match the training image and the CT model perfectly.

Application of three-dimensional image reconstruction technology based on high-resolution CT in pyeloplasty.

BACKGROUND: Three-dimensional (3D) image reconstruction technology is widely used in surgical operations for its intuitive visualization. Pyeloplasty requiresprecise cutting and suturing. The reconstruction technology can accurately determine the location and scope of the stenosis at the junction of the renal pelvis and ureter and the relationship with the surrounding vasculature. The purpose of this article is to retrospective evaluate the application value of image reconstruction technology in pyeloplasty based on high-resolution 3D CT images. METHODS: A total of 20 patients with renal pelvic ureteral junction obstruction admitted to our hospital from August 2019 to August 2020 were selected. In this group, left pyeloplasty was performed in 8 patients and right pyeloplasty in 12 patients. In terms of conditions, there was 1 case with secondary pyeloplasty, 6 cases of patients with kidney stones, 2 cases with renal ectopic blood vessels, 1 case with renal prolapse, 1 case with horseshoe kidney, and 1 case with ureteral polyps. There were 12 males and 8 females, with an average age of 34.65±10.67 years and an average body mass index (BMI) of 22.48±3.03 kg/m2. In all patients, 3D CT reconstruction technology was used to guide the formulation of robot-assisted laparoscopic pyeloplasty plans; verify the consistency between the actual operation and the preoperative planning; and observe the operation time, blood loss, postoperative exhaust time, indwelling drainage tube time, and follow-up for comorbidities. RESULTS: The operation was successful in all 20 patients. The actual operation was 100% consistent with the preoperative planning, the operative time was 160.80±63.26 min, the intraoperative blood loss was 47±30.45 mL, the postoperative exhaust time was 1.15±0.37 days, the drainage tube indwelling time was 4.35±1.50 days, and the average follow-up time was 7.95±3.41 months. There were no complications. CONCLUSIONS: Three-dimensional image reconstruction technology based on high-resolution CT has high clinical application value in the treatment of ureteropelvic junction obstruction (UPJO), which simplifies the operation process and shortens the operation time, and is a valuable tool for auxiliary surgeons in devising the operation plan.

Outcomes of surgery for giant hepatic hemangioma.

BACKGROUND: The surgical indications for liver hemangioma remain unclear. METHODS: Data from 152 patients with hepatic hemangioma who underwent hepatectomy between 2004 and 2019 were retrospectively reviewed. We analyzed characteristics including tumor size, surgical parameters, and variables associated with Kasabach-Merritt syndrome and compared the outcomes of laparoscopic and open hepatectomy. Here, we describe surgical techniques for giant hepatic hemangioma and report on two meaningful cases. RESULTS: Most (63.8%) patients with hepatic hemangioma were asymptomatic. Most (86.4%) tumors from patients with Kasabach-Merritt syndrome were larger than 15 cm. Enucleation (30.9%), sectionectomy (28.9%), hemihepatectomy (25.7%), and the removal of more than half of the liver (14.5%) were performed through open (87.5%) and laparoscopic (12.5%) approaches. Laparoscopic hepatectomy is associated with an operative time, estimated blood loss, and major morbidity and mortality rate similar to those of open hepatectomy, but a shorter length of stay. 3D image reconstruction is an alternative for diagnosis and surgical planning for partial hepatectomy. CONCLUSION: The main indication for surgery is giant (> 10 cm) liver hemangioma, with or without symptoms. Laparoscopic hepatectomy was an effective option for hepatic hemangioma treatment. For extremely giant hemangiomas, 3D image reconstruction was indispensable. Hepatectomy should be performed by experienced hepatic surgeons.

Parvovirus Capsid-Antibody Complex Structures Reveal Conservation of Antigenic Epitopes Across the Family.

The parvoviruses are small nonenveloped single stranded DNA viruses that constitute members that range from apathogenic to pathogenic in humans and animals. The infection with a parvovirus results in the generation of antibodies against the viral capsid by the host immune system to eliminate the virus and to prevent re-infection. For members currently either being developed as delivery vectors for gene therapy applications or as oncolytic biologics for tumor therapy, efforts are aimed at combating the detrimental effects of pre-existing or post-treatment antibodies that can eliminate therapeutic benefits. Therefore, understanding antigenic epitopes of parvoviruses can provide crucial information for the development of vaccination applications and engineering novel capsids able to escape antibody recognition. This review aims to capture the information for the binding regions of ∼30 capsid-antibody complex structures of different parvovirus capsids determined to date by cryo-electron microscopy and three-dimensional image reconstruction. The comparison of all complex structures revealed the conservation of antigenic regions among parvoviruses from different genera despite low sequence identity and indicates that the available data can be used across the family for vaccine development and capsid engineering.

An automatic electronic instrument for accurate measurements of food volume and density.

OBJECTIVE: Accurate measurements of food volume and density are often required as 'gold standards' for calibration of image-based dietary assessment and food database development. Currently, there is no specialised laboratory instrument for these measurements. We present the design of a new volume of density (VD) meter to bridge this technological gap. DESIGN: Our design consists of a turntable, a load sensor, a set of cameras and lights installed on an arc-shaped stationary support, and a microcomputer. It acquires an array of food images, reconstructs a 3D volumetric model, weighs the food and calculates both food volume and density, all in an automatic process controlled by the microcomputer. To adapt to the complex shapes of foods, a new food surface model, derived from the electric field of charged particles, is developed for 3D point cloud reconstruction of either convex or concave food surfaces. RESULTS: We conducted two experiments to evaluate the VD meter. The first experiment utilised computer-synthesised 3D objects with prescribed convex and concave surfaces of known volumes to investigate different food surface types. The second experiment was based on actual foods with different shapes, colours and textures. Our results indicated that, for synthesised objects, the measurement error of the electric field-based method was <1 %, significantly lower compared with traditional methods. For real-world foods, the measurement error depended on the types of food volumes (detailed discussion included). The largest error was approximately 5 %. CONCLUSION: The VD meter provides a new electronic instrument to support advanced research in nutrition science.

Treatment of Giant Left Atrial Diverticulum: A Case Report and Literature Review.

In this paper, we present a giant left atrial diverticulum (LAD) in a 10-year-old girl, whose three-dimensional (3D) image reconstruction was used to help diagnosis and surgical positioning. Previously reported cases were reviewed, and the clinical characteristics of this disease also was summarized to improve the diagnosis and treatment of LAD.

3D Reconstruction of Slug Flow in Mini-Channels with a Simple and Low-Cost Optical Sensor.

The present work provides a new approach for 3D image reconstruction of gas-liquid two-phase flow (GLF) in mini-channels based on a new optical sensor. The sensor consists of a vertical and a horizontal photodiode array. Firstly, with the optical signals obtained by the vertical array, a measurement model developed by Support Vector Regression (SVR) was used to determine the cross-sectional information. The determined information was further used to reconstruct cross-sectional 2D images. Then, the gas velocity was calculated according to the signals obtained by the horizontal array, and the spatial interval of the 2D images was determined. Finally, 3D images were reconstructed by piling up the 2D images. In this work, the cross-sectional gas-liquid interface was considered as circular, and high-speed visualization was utilized to provide the reference values. The image deformation caused by channel wall was also considered. Experiments of slug flow in a channel with an inner diameter of 4.0 mm were carried out. The results verify the feasibility of the proposed 3D reconstruction method. The proposed method has the advantages of simple construct, low cost, and easily multipliable. The reconstructed 3D images can provide detailed and undistorted information of flow structure, which could further improve the measurement accuracy of other important parameters of gas-liquid two-phase flow, such as void fraction, pressure drop, and flow pattern.

Integrated virtual and cadaveric dissection laboratories enhance first year medical students' anatomy experience: a pilot study.

BACKGROUND: Radiology integration into medical anatomy courses is well established, but there is a paucity of literature on integrating virtual dissection into cadaveric dissection laboratories. Virtual dissection is the digital dissection of medical images on touchscreen anatomy visualization tables. The purpose of this pilot study was to investigate the feasibility of integrating virtual dissection into a first-year medical cadaver-based anatomy course and to assess students' overall attitude towards this new technology. METHODS: All students in first-year medicine at a single medical school participated in this study (n = 292). Six virtual dissection laboratories, which focused on normal anatomy, were developed and integrated into a cadaver-based anatomy course. The virtual dissection table (VDT) was also integrated into the final anatomy spot exam. Following the course, students completed a short evidence-informed survey which was developed using a theoretical framework for curriculum evaluation. Numerical data were tabulated, and qualitative content analysis was performed on students' unstructured comments. RESULTS: The survey response rate was 69.2% (n = 202/292). Most (78.7%) students reported that virtual dissection enhanced their understanding of the cadaveric anatomy and the clinical applications of anatomy. Most (73.8%) students also felt that the VDT was an effective use of the laboratory time. Thirteen narrative comments were collected, most of which (61.5%) identified strengths of the curriculum. CONCLUSIONS: In this pilot study, students perceived that their learning was enhanced when virtual dissection was combined with a cadaver-based anatomy laboratory. This study demonstrates that there is potential for virtual dissection to augment cadaveric dissection in medical education.

Optical imaging and modulation of neurovascular responses.

The cerebral microvasculature consists of pial vascular networks, parenchymal descending arterioles, ascending venules and parenchymal capillaries. This vascular compartmentalization is vital to precisely deliver blood to balance continuously varying neural demands in multiple brain regions. Optical imaging techniques have facilitated the investigation of dynamic spatial and temporal properties of microvascular functions in real time. Their combination with transgenic animal models encoding specific genetic targets have further strengthened the importance of optical methods for neurovascular research by allowing for the modulation and monitoring of neuro vascular function. Image analysis methods with three-dimensional reconstruction are also helping to understand the complexity of microscopic observations. Here, we review the compartmentalized cerebral microvascular responses to global perturbations as well as regional changes in response to neural activity to highlight the differences in vascular action sites. In addition, microvascular responses elicited by optical modulation of different cell-type targets are summarized with emphasis on variable spatiotemporal dynamics of microvascular responses. Finally, long-term changes in microvascular compartmentalization are discussed to help understand potential relationships between CBF disturbances and the development of neurodegenerative diseases and cognitive decline.

Application of 3D image reconstruction in robotic urological surgery / 中华泌尿外科杂志

Objective To evaluate the clinical value of three-dimensiomal (3D) image reconstruction in the urologic robot-assisted laparoscopic surgery.Methods From Sept.2017 to Jun.2018,7 patients (3 male cases,4 female cases) diagnosed with genitourinary tumors in Beijing United Family Hospital were involved in this study.The median age was 55 years(IQR 36-64 years).There were 1 case of renal tumor,1 case of adrenal gland tumor,1 case of retroperitoneal tumor,1 case of metastatic tumor post-nephrectomy,1 case of bladder neuroendocrine tumor and 1 case of urachal tumor.All the patients underwent four-phase enhanced CT scan and the CT images were imported into 3D visualization system for 3D image reconstruction,aiming to assist surgery planning and tumor locating.The main parameters of 3D image reconstruction included tumor size,location and the relationships among tumor,organs and blood vessels.All the 7 patients underwent robot-assisted laparoscopic surgeries,including 2 cases of partial nephrectomy (1 case with no blockage of renal artery),1 case of excision of right adrenal gland tumor,1 case of transperitoneal excision of left renal hilus chromaffinoma,1 case of transperitoneal excision of right adrenal gland and postcaval metastatic tumors,1 case of transperitoneal radical cystectomy + orthotopic bladder substitution + extended pelvic lymph node dissection,1 case of transperitoneal excision of urachal tumor + partial cystectomy + pelvic lymph node dissection.Results 3D image reconstruction svstem allows accurate evaluation of tumors,related vessels,surrounding tissue and organs before operation.All the 7 cases surgeries were uneventful,no case was converted to open or conventional laparoscopic surgery,the median operation time was 225 min (IQR 135-280 min),the median blood loss was 50 ml (IQR 20-600 ml).One patient had fever post-operation and there was no over Clavien Dindo Ⅱ perioperative complication.Conclusions 3D image reconstruction shows the great clinical value in approach selection,reduction of tissue and organ damage,reduction of surgery related complication and improving the success rate in robot-assisted laparoscopic surgery.

Estimated functional renal parenchymal volume predicts the split renal function following renal surgery.

PURPOSE: The change in functional renal volume (FRV) has an absolute influence on renal function after nephrectomy (Nx) or nephron-sparing surgery (NSS). In this study, we prospectively examined whether the postoperative renal function following Nx and NSS could be accurately predicted and assessed the reproducibility of our newly developed 3-D image reconstruction system (Kashihara) to measure the FRV. METHODS: We enrolled 98 patients who underwent Nx and 41 patients who underwent NSS from April 2006 to September 2009 to predict postoperative FRV and renal function. FRV was measured before and after (1 month and 1 year) renal surgery. The postoperative estimated glomerular filtration rate (eGFR) was predicted from the preoperative eGFR calculated from the serum creatinine (sCr) level and the ratio of the postoperative/preoperative FRV. To assess the reproducibility and accuracy of our newly developed 3-dimensional (3-D) image reconstruction system, FRV was measured by five examiners using images obtained by CT (five cases) and MRI (five cases). RESULTS: Significant correlation was found both for FRV and for renal function between the predictive values and the actually measured values at 1 month and 1 year after surgery, not only in the Nx group, but also in the NSS group. The accuracy and reproducibility could be confirmed both with CT and MRI studies. CONCLUSIONS: The postoperative FRV and renal function could be predicted preoperatively using a 3-D image reconstructive system, preoperative routine diagnostic imaging, and preoperative sCr level.

Study on the Structure of Candida Albicans-Staphylococcus Epidermidis Mixed Species Biofilm on Polyvinyl Chloride Biomaterial.

The objective of the study was to establish an in vitro model of Candida albicans-Staphylococcus epidermidis mixed species biofilm (BF) on polyvinyl chloride (PVC) material, and to investigate the formation and the structure of mixed species BF formation using a combined approach of confocal laser scanning microscope (CLSM), scanning electron microscope (SEM), and 3D image reconstruction technique. Mixed species BF is achieved by co-incubating Staphylococcus epidermidis bacteria (ATCC35984) and Candida albicans fungal (ATCC10231) with PVC pieces in Tris-buffered saline. BF formation was examined at 2, 6, 12, 24, 48, and 72 h of co-culture. Thickness of these BFs and the number, and percentage of viable cells in BFs were measured. CT scan images of BFs were obtained using CLSM and SEM and reconstructed 3D images of mixed species BF were acquired, in an effort to examine structure of the BF. Staphylococcus epidermidis attached to various forms of candida albicans (spores, pseudohyphae, and hyphae), formed complex and dense mesh arrays. The BF is constituted of a large number of viable and dead pathogens, the surface of mixed species BF is uneven, with living pathogens predominating protrusive portions and dead pathogens aggregating in concaves. Mixed species BF formation on the surface of PVC material was found to be a dynamic process, with rapid growth being at 24 h of co-culture, maximal thickness peaked at 48 h. These mixed species BF matured at 48-72 h. Significant differences were observed in the proportion of viable cells between interior, middle, and outer layers of BFs (p < 0.05). Mixed species BF Candida albicans-Staphylococcus epidermidis is sophisticated in structure. The combined approach involving CLSM, SEM, and 3D image reconstruction technique is ideal for the investigation of mixed species BF on PVC material.