Temporal-Focusing Multiphoton Excitation Single-Molecule Localization Microscopy Using Spontaneously Blinking Fluorophores.

Single-molecule localization microscopy (SMLM) based on temporal-focusing multiphoton excitation (TFMPE) and single-wavelength excitation is used to visualize the three-dimensional (3D) distribution of spontaneously blinking fluorophore-labeled subcellular structures in a thick specimen with a nanoscale-level spatial resolution. To eliminate the photobleaching effect of unlocalized molecules in out-of-focus regions for improving the utilization rate of the photon budget in 3D SMLM imaging, SMLM with single-wavelength TFMPE achieves wide-field and axially confined two-photon excitation (TPE) of spontaneously blinking fluorophores. TPE spectral measurement of blinking fluorophores is then conducted through TFMPE imaging at a tunable excitation wavelength, yielding the optimal TPE wavelength for increasing the number of detected photons from a single blinking event during SMLM. Subsequently, the TPE fluorescence of blinking fluorophores is recorded to obtain a two-dimensional TFMPE-SMLM image of the microtubules in cancer cells with a localization precision of 18 ± 6 nm and an overall imaging resolution of approximately 51 nm, which is estimated based on the contribution of Nyquist resolution and localization precision. Combined with astigmatic imaging, the system is capable of 3D TFMPE-SMLM imaging of brain tissue section of a 5XFAD transgenic mouse with the pathological features of Alzheimer's disease, revealing the distribution of neurotoxic amyloid-beta peptide deposits.

CellSNAP: a fast, accurate algorithm for 3D cell segmentation in quantitative phase imaging.

Significance: Three-dimensional quantitative phase imaging (QPI) has rapidly emerged as a complementary tool to fluorescence imaging, as it provides an objective measure of cell morphology and dynamics, free of variability due to contrast agents. It has opened up new directions of investigation by providing systematic and correlative analysis of various cellular parameters without limitations of photobleaching and phototoxicity. While current QPI systems allow the rapid acquisition of tomographic images, the pipeline to analyze these raw three-dimensional (3D) tomograms is not well-developed. We focus on a critical, yet often underappreciated, step of the analysis pipeline that of 3D cell segmentation from the acquired tomograms. Aim: We report the CellSNAP (Cell Segmentation via Novel Algorithm for Phase Imaging) algorithm for the 3D segmentation of QPI images. Approach: The cell segmentation algorithm mimics the gemstone extraction process, initiating with a coarse 3D extrusion from a two-dimensional (2D) segmented mask to outline the cell structure. A 2D image is generated, and a segmentation algorithm identifies the boundary in the x-y plane. Leveraging cell continuity in consecutive z-stacks, a refined 3D segmentation, akin to fine chiseling in gemstone carving, completes the process. Results: The CellSNAP algorithm outstrips the current gold standard in terms of speed, robustness, and implementation, achieving cell segmentation under 2 s per cell on a single-core processor. The implementation of CellSNAP can easily be parallelized on a multi-core system for further speed improvements. For the cases where segmentation is possible with the existing standard method, our algorithm displays an average difference of 5% for dry mass and 8% for volume measurements. We also show that CellSNAP can handle challenging image datasets where cells are clumped and marred by interferogram drifts, which pose major difficulties for all QPI-focused AI-based segmentation tools. Conclusion: Our proposed method is less memory intensive and significantly faster than existing methods. The method can be easily implemented on a student laptop. Since the approach is rule-based, there is no need to collect a lot of imaging data and manually annotate them to perform machine learning based training of the model. We envision our work will lead to broader adoption of QPI imaging for high-throughput analysis, which has, in part, been stymied by a lack of suitable image segmentation tools.

New method for analysing spatial relationships of facial muscles on MRI: a pilot study.

Dysfunction of the facial musculature can have significant physical, social, and psychological consequences. In surgeries such as cleft surgery or craniofacial bimaxillary osteotomies, the perioral facial muscles may be detached or severed, potentially altering their functional vectors and mimicry capabilities. Ensuring correct reconstruction and maintenance of anatomical sites and muscle vectors is crucial in these procedures. However, a standardized method for perioperative assessment of the facial musculature and function is currently lacking. The aim of this study was to develop a workflow to analyse the three-dimensional vectors of the facial musculature using magnetic resonance imaging (MRI) scans. A protocol for localizing the origins and insertions of these muscles was established. The protocol was implemented using the 3DMedX computer program and tested on 7 Tesla MRI scans obtained from 10 healthy volunteers. Inter- and intra-observer variability were assessed to validate the protocol. The absolute intra-observer variability was 2.6 mm (standard deviation 2.0 mm), and absolute inter-observer variability was 2.6 mm (standard deviation 1.5 mm). This study presents a reliable and reproducible method for analysing the spatial relationships and functional significance of the facial muscles. The workflow developed facilitates perioperative assessment of the facial musculature, potentially aiding clinicians in surgical planning and potentially enhancing the outcomes of midface surgery.

Skeletal changes after midface surgery in patients with craniofacial deformities: a three-dimensional quantification method.

To determine the skeletal changes after midface surgery in patients with syndromic craniosynostosis who underwent Le Fort III (LFIII), monobloc (MB), or facial bipartition (FB). This was a retrospective study including 75 patients: 33 treated by LFIII, 29 by MB, and 13 by FB. Twenty-five had a diagnosis of Apert, 39 Crouzon, and 11 craniofrontonasal syndrome. A three-dimensional mesh was created from the preoperative scan and registered to the postoperative scan to visualise the advancement. LFIII at age 7-12 years effectuated a higher mean advancement in the maxillary (15.5 mm) and zygomatic (7.6 mm) regions when compared to ≥13 years (10.2 mm and 5.5 mm). After MB, mean advancement of the fronto-orbital region was higher at <7 years (16.4 mm), and similarly lower at ages 7-12 (13.8 mm) and ≥13 (12.5 mm). The mean preoperative inter-dacryon distance (34.4 ± 4.4 mm) was reduced by 8.7 ± 4.2 mm after FB without distraction (n = 10). More advancement was seen when midface surgery was performed at a younger age, due to more severe cases and a desire for overcorrection. The highest mean advancement was observed in the fronto-orbital region. Antero-inferior rotational movement was seen after all three techniques.

Three-Dimensional Volumetric Investigation of Onodi Cells: A Multi-Slice Computed Tomography Study.

Introduction Onodi cells (OCs) are posterior ethmoid cells that are located above the sphenoid sinus, close to or even surrounding the carotid artery and optic nerve. Objective To investigate and evaluate the volumetric variation of OCs through multi-slice computed tomography (MSCT) scans. Methods We performed a retrospective review of MSCT scans of 79 subjects, 40 male and 39 female patients, Whose age ranged from 18 to 83 (mean: 39.6) years. The volumes of the OCs on the right and left sides were measured using the ITK-SNAP software (open-source) with semiautomatic segmentation. The possible relationships involving age, gender, contact with the optic nerve, extension of the pneumatization of the posterior ethmoid cells into the clinoid processes, mucous thickening in the anterior and posterior ethmoid cells, and obliteration of the sphenoethmoidal complex were analyzed with the Pearson correlation and Chi-squared tests according to the type of data compared and logistic regression models ( p < 0.05). Results We observed that an increase of one unit in the volume of OCs also increases the chance of extension of pneumatization into the clinoid processes by 0.15% ( p = 0.001). No significant correlations were identified regarding age, gender, and volume of the OCs. Conclusion The volume of the OCs has effects on the extension of pneumatization into the clinoid processes.

Digital setup accuracy for moderate crowding correction with fixed orthodontic appliances: a prospective study.

OBJECTIVES: To evaluate the accuracy of a semi-automatic 3D digital setup process in predicting the orthodontic treatment outcome achieved by labial fixed appliances. SUBJECTS AND METHODS: Twenty-five adult patients (18 to 24 years old) with class I malocclusion and moderate crowding were prospectively enrolled and received treatment on both jaws through the straight-wire technique. Prior to treatment commencement, a semi-automatic digital setup simulating the predicted treatment outcome was performed for each patient through Orthoanalyzer software (3Shape®, Copenhagen, Denmark) to obtain the prediction model. This was compared to the final outcome model through 3D superimposition methods. Metric variables and inspection of color-coded distance maps were used to detect how accurately the digital setup predicts the actual treatment outcome. RESULTS: The mean absolute distances (MAD) between the superimposed dental arches of the predicted and the final models were: 0.77 ± 0.13 mm following superimposition on the palate, 0.52 ± 0.06 mm following superimposition on the maxillary dental arch, and 0.55 ± 0.15 mm following superimposition on the mandibular dental arch. The MAD at the palatal reference area was 0.09 ± 0.04 mm. Visualization of color-coded distance maps indicated that the digital setup accurately predicted the final teeth position in a few cases. Almost half of the cases had posteriorly wider upper and lower dental arches and palatally/lingually positioned or inclined anterior teeth, whereas the rest still showed errors within 2-3 mm, distributed over the entire dental arches with no distinct pattern. CONCLUSIONS: The accuracy of semi-automatic prediction of the labial fixed appliance treatment outcome in Class I cases with moderate crowding is not yet sufficient. While average measures showed deviations less than 1 mm, examination of individual color-coded distance maps revealed significant disparities between the simulated and the actual results.

The surgery-first approach for orthognathic correction of maxillary deficiency-is it stable? Three-dimensional assessment of CBCT scans and digital dental models.

The aim of this study was to determine the skeletal stability of Le Fort I maxillary advancement following the surgery-first approach, by three-dimensional (3D) assessment of cone beam computed tomography (CBCT) scans and digital dental models. CBCT scans of 25 class III patients obtained 1 week preoperatively (T0) and 1 week (T1) and 6 months (T2) postoperatively were superimposed to measure surgical movements (T0-T1) and skeletal relapse (T1-T2). The distorted dentition of the CBCT scans at T1 was replaced with 3D images of the dental models to assess the postoperative occlusion. Surgical movements of the maxilla (mean ± standard deviation values) were 6.79 ± 2.30 mm advancement, 1.28 ± 1.09 mm vertically, and 0.71 ± 0.79 mm mediolaterally. Horizontal rotation (yaw) was 1.56° ± 1.21°, vertical rotation (pitch) 1.86° ± 1.88°, and tilting (roll) 1.63° ± 1.54°. At T2, the posterior relapse was 0.72 ± 0.43 mm (P = 0.001) and relapse in pitch was 1.56° ± 1.42° (P = 0.007). There was no correlation between the size of the surgical movements and the amount of relapse. A weak correlation was noted between the number of teeth in occlusal contact immediately following surgery and relapse of maxillary roll (r = - 0.434, P = 0.030). The stability of maxillary advancement with the surgery-first approach was satisfactory and was not correlated with the quality of the immediate postoperative occlusion.

Multiscale transport and 4D time-lapse imaging in precision-cut liver slices (PCLS).

Background: Monitoring cellular processes across different levels of complexity, from the cellular to the tissue scale, is important for understanding tissue structure and function. However, it is challenging to monitor and estimate these structural and dynamic interactions within three-dimensional (3D) tissue models. Objective: The aim of this study was to design a method for imaging, tracking, and quantifying 3D changes in cell morphology (shape and size) within liver tissue, specifically a precision-cut liver slice (PCLS). A PCLS is a 3D model of the liver that allows the study of the structure and function of liver cells in their native microenvironment. Methods: Here, we present a method for imaging liver tissue during anisosmotic exposure in a multispectral four-dimensional manner. Three metrics of tissue morphology were measured to quantify the effects of osmotic stress on liver tissue. We estimated the changes in the volume of whole precision cut liver slices, quantified the changes in nuclei position, and calculated the changes in volumetric responses of tissue-embedded cells. Results: During equilibration with cell-membrane-permeating and non-permeating solutes, the whole tissue experiences shrinkage and expansion. As nuclei showed a change in position and directional displacement under osmotic stress, we demonstrate that nuclei could be used as a probe to measure local osmotic and mechanical stress. Moreover, we demonstrate that cells change their volume within tissue slices as a result of osmotic perturbation and that this change in volume is dependent on the position of the cell within the tissue and the duration of the exposure. Conclusion: The results of this study have implications for a better understanding of multiscale transport, mechanobiology, and triggered biological responses within complex biological structures.

Analyzing Vocal Fold Frequency Dynamics Using High-Speed 3D Laser Video Endoscopy.

OBJECTIVE: To examine changes in lateral and vertical vibratory motion along the anterior, middle, and posterior sections of the vocal folds, as a function of vocal frequency variations. METHODS: Absolute measurements of vocal fold surface dynamics from high-speed videoendoscopy with custom laser endoscope were made on 23 vocally healthy adults during sustained /i:/ production at 10%, 20%, and 80% of pitch range. The 3D parameters of amplitude (mm), maximum velocity opening/closing (mm/s), and mean velocity opening/closing (mm/s) were computed for the lateral and vertical vibratory motion along the anterior, middle, and posterior sections of the vocal folds. Linear mixed model analysis was conducted to evaluate the differences in (a) vocal frequency levels (high vs. normal vs. low pitch), (b) axis level (vertical vs. lateral), (c) position level (anterior vs. middle vs. posterior), and (d) gender differences (male vs. female). RESULTS: Overall, the superior surface vertical motion of the vocal fold is greater compared with the lateral motion, especially in males. Along the superior surface, the mean and maximum closing velocities are greater posteriorly for low pitch. The location (anterior, middle, and posterior) along the superior surface is relevant only for vocal fold closing rather than opening, as the dynamics are different along the various locations. CONCLUSIONS: The study highlights the significance of assessing the vertical motion of the superior surface of the vocal fold to understand the complex dynamics of voice production. LEVEL OF EVIDENCE: NA Laryngoscope, 2024.

Automatic 3D Augmented-Reality Robot-Assisted Partial Nephrectomy Using Machine Learning: Our Pioneer Experience.

The aim of "Precision Surgery" is to reduce the impact of surgeries on patients' global health. In this context, over the last years, the use of three-dimensional virtual models (3DVMs) of organs has allowed for intraoperative guidance, showing hidden anatomical targets, thus limiting healthy-tissue dissections and subsequent damage during an operation. In order to provide an automatic 3DVM overlapping in the surgical field, we developed and tested a new software, called "ikidney", based on convolutional neural networks (CNNs). From January 2022 to April 2023, patients affected by organ-confined renal masses amenable to RAPN were enrolled. A bioengineer, a software developer, and a surgeon collaborated to create hyper-accurate 3D models for automatic 3D AR-guided RAPN, using CNNs. For each patient, demographic and clinical data were collected. A total of 13 patients were included in the present study. The average anchoring time was 11 (6-13) s. Unintended 3D-model automatic co-registration temporary failures happened in a static setting in one patient, while this happened in one patient in a dynamic setting. There was one failure; in this single case, an ultrasound drop-in probe was used to detect the neoplasm, and the surgery was performed under ultrasound guidance instead of AR guidance. No major intraoperative nor postoperative complications (i.e., Clavien Dindo > 2) were recorded. The employment of AI has unveiled several new scenarios in clinical practice, thanks to its ability to perform specific tasks autonomously. We employed CNNs for an automatic 3DVM overlapping during RAPN, thus improving the accuracy of the superimposition process.

Optical Three-Dimensional Imaging for Objective Evaluation of the Donor Site after Anterolateral Thigh Flap Surgery.

Background: The high volume of the fasciomyocutaneous anterolateral thigh flaps (ALT) is suitable for the reconstruction of pronounced soft tissue defects. At the same time, harvesting ALT results in a drastic change in thigh shape. Here, we present an optical three-dimensional imaging method for thigh comparison, which can be an objective and reproducible method for evaluating donor sites after ALT harvesting. Methods: In total, 128 thighs were scanned with an optical three-dimensional scanner, Vectra XT ®. Sixty-eight non-operated right and left thighs were compared and served as a control. Sixty thighs were scanned in the ALT group. The average surface area deviations, thigh volume, thigh circumference, and flap ratio to thigh circumference were calculated. The results were correlated with Δthigh circumference and Δvolume of the unoperated thighs of the control group. Results: No significant difference between the thigh volumes of the right and left thighs was found in the control group. Removal of an ALT flap showed a significant (p < 0.007) volume reduction compared to unoperated thighs (2.7 ± 0.8 L and 3.3 ± 0.9 L, respectively). Flap area correlated strongly with the Δthigh circumference (r = 0.66, p < 0.001) and Δvolume (r = 0.68, p < 0.001). Strong correlations were observed between flap ratio and thigh circumference with Δhigh circumference (r = 0.57, p < 0.001) and Δvolume (r = 0.46, p < 0.05). Conclusions: Optical three-dimensional imaging provides an objective and reproducible tool for detecting changes in thigh morphology volume differences after ALT harvesting.

Impact of intraoperative imaging on decision-making during spine surgery: a survey among spine surgeons using simulated intraoperative images.

PURPOSE: To assess whether the intention to intraoperatively reposition pedicle screws differs when spine surgeons evaluate the same screws with 2D imaging or 3D imaging. METHODS: In this online survey study, 21 spine surgeons evaluated eight pedicle screws from patients who had undergone posterior spinal fixation. In a simulated intraoperative setting, surgeons had to decide if they would reposition a marked pedicle screw based on its position in the provided radiologic imaging. The eight assessed pedicle screws varied in radiologic position, including two screws positioned within the pedicle, two breaching the pedicle cortex < 2 mm, two breaching the pedicle cortex 2-4 mm, and two positioned completely outside the pedicle. Surgeons assessed each pedicle screw twice without knowing and in random order: once with a scrollable three-dimensional (3D) image and once with two oblique fluoroscopic two-dimensional (2D) images. RESULTS: Almost all surgeons (19/21) intended to reposition more pedicle screws based on 3D imaging than on 2D imaging, with a mean number of pedicle screws to be repositioned of, respectively, 4.1 (± 1.3) and 2.0 (± 1.3; p < 0.001). Surgeons intended to reposition two screws placed completely outside the pedicle, one breaching 2-4mm, and one breaching < 2 mm more often based on 3D imaging. CONCLUSION: When provided with 3D imaging, spine surgeons not only intend to intraoperatively reposition pedicle screws at risk of causing postoperative complications more often but also screws with acceptable positions. This study highlights the potential of intraoperative 3D imaging as well as the need for consensus on how to act on intraoperative 3D information.

Three-Dimensional Image Visualization under Photon-Starved Conditions Using N Observations and Statistical Estimation.

In this paper, we propose a method for the three-dimensional (3D) image visualization of objects under photon-starved conditions using multiple observations and statistical estimation. To visualize 3D objects under these conditions, photon counting integral imaging was used, which can extract photons from 3D objects using the Poisson random process. However, this process may not reconstruct 3D images under severely photon-starved conditions due to a lack of photons. Therefore, to solve this problem, in this paper, we propose N-observation photon-counting integral imaging with statistical estimation. Since photons are extracted randomly using the Poisson distribution, increasing the samples of photons can improve the accuracy of photon extraction. In addition, by using a statistical estimation method, such as maximum likelihood estimation, 3D images can be reconstructed. To prove our proposed method, we implemented the optical experiment and calculated its performance metrics, which included the peak signal-to-noise ratio (PSNR), structural similarity (SSIM), peak-to-correlation energy (PCE), and the peak sidelobe ratio (PSR).

Novel evaluation method for facial nerve palsy using 3D facial recognition system in iPhone.

OBJECTIVE: While subjective methods like the Yanagihara system and the House-Brackmann system are standard in evaluating facial paralysis, they are limited by intra- and inter-observer variability. Meanwhile, quantitative objective methods such as electroneurography and electromyography are time-consuming. Our aim was to introduce a swift, objective, and quantitative method for evaluating facial movements. METHODS: We developed an application software (app) that utilizes the facial recognition functionality of the iPhone (Apple Inc., Cupertino, USA) for facial movement evaluation. This app leverages the phone's front camera, infrared radiation, and infrared camera to provide detailed three-dimensional facial topology. It quantitatively compares left and right facial movements by region and displays the movement ratio of the affected side to the opposite side. Evaluations using the app were conducted on both normal and facial palsy subjects and were compared with conventional methods. RESULTS: Our app provided an intuitive user experience, completing evaluations in under a minute, and thus proving practical for regular use. Its evaluation scores correlated highly with the Yanagihara system, the House-Brackmann system, and electromyography. Furthermore, the app outperformed conventional methods in assessing detailed facial movements. CONCLUSION: Our novel iPhone app offers a valuable tool for the comprehensive and efficient evaluation of facial palsy.

Evaluating the benefit of virtual surgical planning on bony union rates in head and neck reconstructive surgery.

OBJECTIVES: Virtual surgical planning (VSP) has gained acceptance because of its benefits in obtaining adequate resection, achieving cephalometric accuracy, and reducing operative time. The aim of this study is to compare the rate of union between VSP and free-hand surgery (FHS), identify predictors of non-union and evaluate the difference in operative time. METHODS: Post-operative CT were retrospectively reviewed for 123 patients who underwent maxillary or mandibular reconstruction between 2014 and 2021 using either VSP or FHS. Each apposition was graded as complete, partial or non-union. The rate of union, risk difference and inter-rater reliability were calculated. The difference in operative time was assessed. Predictors of non-union were identified using logistic regression. RESULTS: A total of 326 appositions were graded (VSP n = 150; FHS n = 176). The rates of complete and partial union were higher with VSP than FHS (74.7% vs. 65.3%; 18% vs. 15.9%, respectively, p = 0.01). Non-union was found at a higher rate with FHS than with VSP (18.7% vs. 7.3%). The non-union risk difference was 11.4. FHS, major complications and apposition at the native bone were predictors of non-union (OR 2.9, p = 0.02; OR 3.4, p = 0.01; OR 2.5, p = 0.05, respectively). The mean surgical time was shorter with VSP than with FHS (265.3 vs. 381.5 min, p < 0.001). The inter-rater agreement was high (k = 0.85; ICC = 0.86). CONCLUSION: VSP demonstrated significantly higher bony union rates and shorter operative time. FHS, development of major complications and apposition with native bone correlated with non-union.

In Vivo Intelligent Fluorescence Endo-Microscopy by Varifocal Meta-Device and Deep Learning.

Endo-microscopy is crucial for real-time 3D visualization of internal tissues and subcellular structures. Conventional methods rely on axial movement of optical components for precise focus adjustment, limiting miniaturization and complicating procedures. Meta-device, composed of artificial nanostructures, is an emerging optical flat device that can freely manipulate the phase and amplitude of light. Here, an intelligent fluorescence endo-microscope is developed based on varifocal meta-lens and deep learning (DL). The breakthrough enables in vivo 3D imaging of mouse brains, where varifocal meta-lens focal length adjusts through relative rotation angle. The system offers key advantages such as invariant magnification, a large field-of-view, and optical sectioning at a maximum focal length tuning range of ≈2 mm with 3 µm lateral resolution. Using a DL network, image acquisition time and system complexity are significantly reduced, and in vivo high-resolution brain images of detailed vessels and surrounding perivascular space are clearly observed within 0.1 s (≈50 times faster). The approach will benefit various surgical procedures, such as gastrointestinal biopsies, neural imaging, brain surgery, etc.

Relationship between Bilateral Landmarks of Facial Asymmetry in Skeletal Class II and Class III in Vertical Dimension: 3D Facial Scan and Cone-Beam Computed Tomography.

Significant facial asymmetry can lead to both functional and aesthetic issues. Often, such asymmetry originates from irregularities in the jaw structure. It is critical to recognize that asymmetries can be concealed by soft tissue, which may be overlooked. The aim of this study is to investigate the relationships between bilateral landmarks in the vertical dimension of facial asymmetry among individuals with skeletal Class II and Class III malocclusions. Fifty-two adult patients with a mean age of 24.4 ± 3.79 years with facial asymmetry who underwent orthodontic and orthognathic surgery were studied. Cone-beam computed tomography and facial scans were used to create 3D virtual head models which enhanced the accuracy in addressing facial asymmetry to ensure effective treatment planning. Each landmark was measured and digitized using the Dolphin Imaging program. The findings showed a correlation between gender and type of skeletal discrepancies with the menton deviation direction. In conclusion, the vertical discrepancy between bilateral landmarks was observable in both hard and soft tissues with more pronounced expression in soft tissues. This discrepancy was characterized by an elevation on the same side as the menton deviation, which was a feature observed in both skeletal Class II and Class III individuals.

Intramedullary nailing of femoral shaft fractures: an analysis of rotational malunions using 3D EOS.

PURPOSE: Rotational malalignment and leg length discrepancy after intramedullary nailing of femoral shaft are frequent. This study has three objectives: evaluate the rate of femoral rotational malalignment and leg length discrepancy using EOS imaging after antegrade intramedullary nailing of femoral shaft fracture, find a relevant clinical examination to detect malrotation and identified risk factors. METHODS: We performed a retrospective single-centre study between January 2014 and January 2022. Fifty-eight patients were clinically and radiographically assessed at a minimum of three months. RESULTS: The femoral rotation of the operated side was significantly greater by a mean of 15.4° in internal rotation compared to the healthy side. There was no statically significant difference for the femoral length (p = 0.08). CONCLUSION: When using EOS stereography following antegrade intramedullary nailing of post-traumatic diaphyseal femur fractures, a statistically significant difference of more than 15.4° in internal rotation was found for femoral rotation on the operated side compared to the healthy side.Please confirm if the author names are presented accurately and in the correct sequence (given name, middle name/initial, family name). Author 1 Given name: [B. Poirot] Last name [Seynaeve]. Also, kindly confirm the details in the metadata are correct.The last name of the first author was corrected : Given name = B. and last name = Poirot Seynaeve The details in matadata are correct LEVEL OF EVIDENCE: III.