Images Of Sections Research Articles

International Consensus Recommendations of Diagnostic Criteria and Terminologies for Extranodal Extension in Head and Neck Squamous Cell Carcinoma: An HN CLEAR Initiative (Update 1).

Extranodal extension (ENE) increases the risk of recurrence and death in head and neck squamous cell carcinoma (HNSCC) patients and is an indication for treatment escalation. Histopathology forms the mainstay of diagnosing ENE. There is substantial variation in the diagnosis of ENE and related terminology. Harmonising the diagnostic criteria for ENE was identified as a priority by the Head and Neck Consensus Language for Ease of Reproducibility (HN CLEAR) Steering Committee and its global stakeholders. An international working group including 16 head and neck pathologists from eight countries across five continents evaluated whole slide images of haematoxylin and eosin-stained sections depicting potential diagnostic problems through nine virtual meetings to develop consensus guidelines. ENE should be diagnosed only when viable carcinoma extends through the primary lymph node (LN) capsule and directly interacts with the extranodal host environment with or without desmoplastic stromal response. Identifying the original LN capsule and reconstruction of its contour can assist in the detection and assessment of ENE. The term matting is recommended for confluence of two or more nodes due to histologically identifiable tumour extending from one LN to another. Matting constitutes major form of ENE. On the other hand, the terms fusion/adhesion/confluence/conglomeration and other synonyms of adhesion should be limited to confluence due to fibrosis or inflammation without histologically identifiable tumour between involved lymph nodes. Tumour extension along narrow needle tracks or spillage of cyst contents following an FNA do not constitute ENE. The consensus recommendations encompassing the definition of ENE, macroscopic and histologic examination of lymph nodes (LN) and practical guidelines for handling challenging cases are provided.

Spectral Hadamard microscopy with metasurface-based patterned illumination

Abstract Hadamard matrices, composed of mutually orthogonal vectors, are widely used in various applications due to their orthogonality. In optical imaging, Hadamard microscopy has been applied to achieve optical sectioning by separating scattering and background noise from desired signals. This method involves sequential illumination using Hadamard patterns and subsequent image processing. However, it typically requires costly light modulation devices, such as digital micromirror devices (DMDs) or spatial light modulators (SLMs), to generate multiple illumination patterns. In this study, we present spectral Hadamard microscopy based on a holographic matasurface. We noticed that certain patterns repeat within other Hadamard patterns under specific condition, allowing the entire set to be reproduced from a single pattern. This finding suggests that generating a single pattern is sufficient to implement Hadamard microscopy. To demonstrate this, we designed a metasurface to generate an illumination pattern and conducted imaging simulations. Results showed that holographic metasurface-based Hadamard microscopy effectively suppressed scattering signals, resulting in clear fluorescent images. Furthermore, we demonstrated that hyperspectral imaging can be achieved with Hadamard microscopy using dispersive optical elements, as the orthogonality of the Hadamard pattern enables to resolve spectral information. The reconstructed hyperspectral images displayed a color distribution closely matching the synthetic hyperspectral images used as ground truth. Our findings suggest that optical sectioning and hyperspectral imaging can be accomplished without light modulation devices, a capability typically unattainable with standard wide-field microscopes. We showed that sophisticated metasurfaces have the potential to replace and enhance conventional optical components, and we anticipate that this study will contribute to advancements in metasurface-based optical microscopy.

RETRACTION: Decellularized Scaffolds with Double-Layer Aligned Microchannels Induce the Oriented Growth of Bladder Smooth Muscle Cells: Toward Urethral and Ureteral Reconstruction.

Q. Cheng, L. Zhang, J. Zhang, X. Zhou, B. Wu, D. Wang, T. Wei, M. Shafiq, S. Li, D. Zhi, Y. Guan, K. Wang, and D. Kong, "Decellularized Scaffolds with Double-Layer Aligned Microchannels Induce the Oriented Growth of Bladder Smooth Muscle Cells: Toward Urethral and Ureteral Reconstruction," Advanced Healthcare Materials 12, no. 26 (2023): 2300544, https://doi.org/10.1002/adhm.202300544. The above article, published online on 28 August 2023 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the authors; the journal Editor-in-Chief, Uta Goebel; and Wiley-VCH GmbH. The retraction has been agreed upon following an investigation into concerns raised by a third party, which revealed inappropriate duplication of image sections and/or formatting artefacts in Figures 2B, 5A, and 7C, as well as errors in Figure S8 and its quantifications. The authors have stated that these image alterations wereperformed to enhance the visual appearance of the images and do not impact the clinical and scientific findings outlined in the paper. Nevertheless, due to the number and the level of inconsistencies identified in the published figures, the editors considered the conclusions substantially compromised and decided to retract the article.

Prediction of both diffusive and hydraulic conductance in the pore network model extracted from 3D images using deep learning

Abstract Pore network modeling (PNM) with network extracted from 3D images is widely used for studying mass transport in porous media, with its accuracy heavily dependent on the precise determination of hydraulic and diffusive conductance values of local pore pairs. Deep learning (DL) prediction based on true pore geometry offers a promising approach for conductance estimation. However, due to arbitrarily orientation and the lack of explicit inlet and outlet surfaces in 3D extracted pore pair geometries, existing studies [24,25] have relied only on 2D cross section images for hydraulic conductance prediction. A custom finite difference method was recently developed, capable exclusively of estimating diffusion conductance from 3D pore geometry [26]. In this work, a convolutional neural network (CNN) was trained to estimate both diffusive and hydraulic conductance values, with reference ground truth values obtained by applying the Lattice Boltzmann Method (LBM) on 3D pore pair geometries. The predicted pore pair conductance values from the CNN closely matched the LBM results on unseen segmented image data. This study highlights the importance of using the full 3D pore pair geometry for accurate hydraulic conductance evaluation, rather than relying solely on 2D throat cross sections. To evaluate the generality of our method, we calculated the permeability and diffusivity of 3D porous media across a wide range of porosities using PNM with local conductance values predicted by our DL model. Comparisons with results of LBM indicate that while diffusivity was accurately predicted, permeability was underestimated. This underestimation is attributed to the over-segmentation of pore spaces, which resulted in an underestimated flow rate.

Pre-screening of endomysial microvessel density by fast random forest image processing machine learning algorithm accelerates recognition of a modified vascular network in idiopathic inflammatory myopathies

Biomarkers for discrimination among different subgroups of idiopathic inflammatory myopathies (IIM) are difficult to identify and may involve multiple laboratory tests and time-consuming procedures. We assessed the potential for artificial intelligence (AI) to extract features such as density of endomysial microvessels based on automatic analysis of the CD31+ vascular network on muscle biopsy images. We also assessed the potential of this technique to save time and its agreement rate with analyses based on the manual selection of microvessels from the same images. A total of 84 images from 84 patients with IIM, diagnosed between 2014 and 2020, were retrieved and analyzed using the Fast Random Forest (FRF) technique. We built a lightweight and explainable algorithm for calculating the pixel percentage of CD31+ endomysial capillaries. The FRF technique applied on images of CD31-stained muscle sections achieved a good performance in the recognition of microvessels by estimating their density over a standard area corresponding to a sample of microscope image. The time spent for this analysis was 90% less than the manual choice of microvessels (estimated time considering the computational time and the time spent to manually detecting the microvessels features). The good performance of the FRF demonstrates that the CD31 pixel percentage of endomysial capillaries is sufficient for a correct estimation. Finally, the paper proposes a procedure to integrate AI in the pre-screening process.

EXPRESSION OF CONCERN: Cannabinoid Receptors as Novel Targets for the Treatment of Melanoma.

C. Blázquez, A. Carracedo, L. Barrado, P. J. Real, J. L. Fernández-Luna, G. Velasco, M. Malumbres, and M. Guzmán, "Cannabinoid Receptors as Novel Targets for the Treatment of Melanoma," The FASEB Journal. 20, no. 14 (2006): 2633-2635, https://doi.org/10.1096/fj.06-6638fje This Expression of Concern is for the above article, published online on 25 October 2006 in Wiley Online Library (wileyonlinelibrary.com), and has been published by agreement between the journal Editor-in-Chief, Loren E. Wold; the Federation of American Societies for Experimental Biology; and Wiley Periodicals LLC. This Expression of Concern has been published due to concerns raised by a third party regarding duplicated image sections and undeclared splicing within the subpanels of Figure1A. The authors acknowledged the image compilation error, but due to the time elapsed since publication, the raw data was not available. The authors stated that their findings regarding melanoma cells expressing functional CB1 and CB2 receptors are supported by other experimental approaches shown in this article, and have been confirmed and published by other independent investigators. Therefore, the authors believe that the irregularities found in Figure1A do not affect the results and the conclusions of this publication. Nevertheless, without an explanation of the anomaly in the figure and in the absence of the original raw data, the journal team could not verify the authenticity of this figure. Therefore, the journal has decided to issue an Expression of Concern to inform and alert the readers.

Ex Vivo Fluorescence Confocal Microscopy for intraoperative evaluations of staple lines and surgical margins in specimens of the lung - a proof-of-concept study.

Intraoperative consultation is frequently used during the surgical treatment of lung tumors for the diagnosis of malignancy and the assessment of surgical margins. The latter is often problematic given the nature of the applied staple lines, which cannot be readily examined in frozen sections. Seventy-nine samples of surgical margins (71 staple lines and 8 open margins) from 52 lung specimens were examined using an ex vivo fluorescence confocal microscope (FCM). The diagnoses of the FCM scans were compared to the corresponding paraffin section images of the same material. The procedure provided intraoperative FCM imaging of the surgical margins and staple lines without having to remove the metal clips. Tumor-involved open margins (5/5) and tumor-involved staple lines (3/4) were correctly identified in the FCM images. The results also provided additional information to the conventional frozen sections. This is the first time staple lines of lung specimens have been visualized as preserved tissue using FCM. The method potentially provides an additional approach for intraoperative decisions when the margins in conventional frozen sections are unclear. Our promising results, however, need to be validated on a larger number of cases.

Long-range inputome of prefrontal GABAergic interneurons in the Alzheimer's disease mouse.

Alzheimer's disease (AD) is the most common neurodegenerative disease, characterized by damage to cortical circuits. However, the mechanisms underlying AD-associated changes in long-range circuits remain poorly understood. In this study, we used viral tracing and fluorescence micro-optical sectioning tomography (fMOST) imaging to investigate whole-brain changes in the input circuit of the frontal cortex of 5×FAD mice. Pathological axonal degeneration was widely observed in upstream regions, including the cortex, hippocampus, and thalamus, across all AD brains examined. The proportion of input neurons projecting to parvalbumin-expressing neurons, compared to those projecting to somatostatin-expressing neurons, decreased in the hippocampus and basal forebrain. This decline was closely related to mouse age and the cell type of the presynaptic input neurons. This study demonstrates the selective vulnerability of long-range circuits in the prelimbic area in AD at the mesoscopic level, thereby enhancing our understanding of circuit architecture degeneration across the brain. We used whole-brain imaging with single-cell resolution to generate brain-wide input maps of the Alzheimer's disease mouse model. The pathological changes in the input proportions showed relevance with the mouse age, distribution, and cell type of the presynaptic input neurons. Compared to the cell body and dendrites of the medial prefrontal cortex input neurons, the pathological changes in the axonal structure are more extensive.

Dissection of the long-range circuit of the mouse intermediate retrosplenial cortex

The retrosplenial cortex (RSP) is a complex brain region with multiple interconnected subregions that plays crucial roles in various cognitive functions, including memory, spatial navigation, and emotion. Understanding the afferent and efferent connectivity of the RSP is essential for comprehending the underlying mechanisms of its functions. Here, via viral tracing and fluorescence micro-optical sectioning tomography (fMOST), we systematically investigated the anatomical organisation of the upstream and downstream circuits of glutamatergic and GABAergic neurons in the dorsal and ventral RSP. The cortical connections of the RSP show laminar organisation in which the input neurons are distributed more in the deeper layers of the upstream cortex. Although different types of neurons have similar upstream circuits, GABAergic neurons show bidirectional connections with the hippocampus, whereas glutamatergic neurons only show unidirectional connections. Moreover, GABAergic neurons receive more inputs from the primary sensory cortex than from the prefrontal cortex and association cortex. The dorsal and ventral subregions have preferred circuits such that the dorsal RSP exhibits spatially topological connections with the dorsal visual cortex and lateral thalamus. The systematic study on long-range connections across RSP subregions and cell types may provide useful information for future revealing of RSP working mechanisms.

Unveiling Microscopic Variations during Photodynamic Therapy via Polarity-Responsive Fluorescence Lifetime Imaging.

Photodynamic therapy is a highly promising method for cancer adjuvant treatment. However, the current research on the microscopic changes during the photodynamic therapy process is still quite limited, which seriously impedes the deep understanding of the procedure. For this purpose, a novel polarity-responsive probe, MI-PPF, with excellent mitochondrial targeting and anchoring capabilities has been rationally designed and synthesized. Notably, MI-PPF has successfully realized the in situ detection of mitochondrial morphology and polarity alterations during the photodynamic therapy process in cancer cells through fluorescence lifetime imaging. The results showed that a series of phenomena such as deformation, shrinkage, vacuolation, and aggregation occurred in the mitochondrial morphology during photodynamic therapy. Concurrently, a decline in mitochondrial polarity is also noted, which may be closely linked to the mitochondrial oxidative stress response during this process. Furthermore, MI-PPF can be used for photodynamic therapy on tumor mouse models and has successfully achieved fluorescence lifetime imaging of tumor sections before and after photodynamic therapy, uncovering multifaceted changes in cell morphology, polarity, and polarity distribution within the mouse tumor model during the process. It is anticipated that this study will offer valuable insights and guidance to the research of mitochondrial-related fields and will boost the advancement of diagnostic and therapeutic areas for associated diseases.

Anatomical and Embryological Development of the Chick Cerebrum in Different Embryonic Periods.

The objective of this study is to assess the embryological and morphometric development of the chick cerebrum during specific incubation periods. The cerebrums of 24 Babcock White Leghorn chicks, six each from the 10th, 13th, 16th and 21st days of the incubation period, were used in the study. After removing the heads of fixed embryos from the upper edge of the atlas, the brains were taken out of the cranial cavity. Morphometric measurements were performed on the removed brains, and paraffin blocks were prepared following the routine histological procedure. Sections 5µm thick were taken from the blocks, with an additional 10µm thick section taken every 50th section. The slides were then stained using Crossmon's triple stain and Klüver-Barrera staining methods and photographed. The sectional images were transferred to the ImageJ programme, brain volume was calculated using stereological methods, and histological measurements were performed. The development of brain parts in selected embryonic periods was examined in detail, focusing on anatomical and histological aspects. According to the results, it was determined that all measured parameters, except the third ventricle width, increased and were statistically significant (p<0.05). It is believed that the findings of this study will enhance the understanding of the region's anatomy. The new morphometric data can serve as reference data in neurotoxicity and embryotoxicity studies.

Assessment of knee cartilage using accelerated 3T MRI: Evaluation of an isotropic 3D fast spin-echo sequence (CUBE) with compressed sensing technique.

Compressed Sensing (CS) is an emerging technique to accelerate MRI acquisitions. The aim of this study was to assess the reliability and accuracy of cartilage thickness measurements in the knee using a CS-enabled isotropic 3D Fast Spin-Echo (FSE) sequence on a 3-T MRI scanner. Twenty-eight tibial condyle sections were collected from 14 adult patients who underwent total knee arthroplasty. An isotropic 3D PDw FSE with variable flip-angle (CUBE) sequence with CS was used to acquire MR images of the tibial condyle sections. Minimum cartilage thickness measurements were independently performed by two experienced readers (R1 and R2) on MR images and compared to corresponding anatomical sections measurements. Intraclass correlation coefficients (ICCs) and Bland-Altman analyses, were used to assess agreement between MR and anatomical measurements. A total of 84 paired cartilage areas were analyzed [cartilage thickness measurements ranged from 0 to 3.40mm at anatomical evaluation (mean, 1.08mm±0.83)]. The agreements between MR and anatomical measurements were excellent (mean differences, 0.06±0.31mm for R1 and 0.03±0.43mm for R2) with respective ICC values of 0.93 and 0.88. Bland-Altman analyses revealed small differences between MR and anatomical measurements, with 95% Limit of Agreements values falling within clinically acceptable ranges (-0.54 to 0.66mm for R1, -0.87 to 0.80mm for R2). The 3D PDw FSE sequence with Compressed Sensing acceleration technique demonstrated reliable and accurate assessment of cartilage thickness of the tibial condyles within a timeframe suitable for routine clinical practice.

Oral delivery of MOMIPP lipid nanoparticles for methuosis-induced cancer chemotherapy.

Methuosis, a non-apoptotic pattern of cell death, triggers the accumulation of macropinosome-derived vacuoles in the cytoplasm. Through this novel mechanism, methuosis inducers possess great potential in fighting apoptosis-resistant cancer cells and offer a promising alternative for cancer treatment. However, the potent methuosis inducer, 3-(5-methoxy, 2-methyl-1H-indol-3-yl)-1-(4-pyridinyl)-2-propen-1-one (MOMIPP), faces an intractable issue of insolubility in most solvents, hindering in vivo dosing and compromising the validation of its antitumor efficacy. Few strategies have been developed to effectively deliver MOMIPP and achieve robust in vivo tumor inhibition since its first report in 2012. Here, a MOMIPP self-emulsifying drug delivery system (MOMIPP-SEDDS) was developed to substantially improve its oral bioavailability and achieve a favorable antitumor effect in a mouse xenograft tumor model. Our findings demonstrated that the MOMIPP-SEDDS was internalized into Caco-2 cells via the lipid raft/caveolae pathway and exhibited enhanced absorption in both cell monolayers and everted gut sacs. Compared with MOMIPP suspensions, MOMIPP-SEDDS showed a 13.3-fold increase in peak concentration and increased relative bioavailability by 19.98 times. By inducing methuosis, MOMIPP-SEDDS successfully retarded tumor progression in a subcutaneous HeLa mouse tumor model. Additionally, transmission electron microscopy (TEM) images of the tumor sections evidenced the occurrence of methuosis in the MOMIPP-SEDDS treatment group. This MOMIPP-SEDDS emerges as a promising lipid nanoparticle platform and high translational medicine for the oral delivery of MOMIPP to exert methuosis-induced tumor suppression for cancer treatment.

Regeneration of a Rabbit Segmental Defect with a New Bone Therapy: Autologous Blood Coagulum with Bone Morphogenetic Protein 6 and Synthetic Ceramics.

Segmental defects of long bones are among the most challenging and debilitating conditions in clinical medicine. Osteogrow-C is a novel osteoinductive device composed of recombinant human bone morphogenetic protein 6 (rhBMP6) delivered within autologous blood coagulum (ABC) with calcium phosphate ceramics that was proven efficacious in preclinical models of spinal fusion. This study aimed to evaluate the efficacy of Osteogrow-C in comparison to that of other osteoinductive therapies in a clinically relevant segmental defect model in rabbits. Segmental defects (15 mm) of rabbit ulna were treated with Osteogrow-C containing different synthetic ceramic particles (tricalcium phosphate [TCP] and TCP/hydroxyapatite 40%/60%), Osteogrow (rhBMP6/ABC), Infuse (rhBMP2/absorbable collagen sponge), and control implants without bone morphogenetic proteins. Defect healing was evaluated by invivo x-ray scans 4, 8, and 17 weeks after the surgery, and animals were killed after 17 weeks for further radiographical and histological assessment. Evaluation of x-ray images, micro-computed tomography, and histological sections revealed that both Osteogrow-C formulations as well as Osteogrow and Infuse promoted healing of the ulnar segmental defect. However, radiographic scores were higher in animals treated with Osteogrow-C than those for the other used therapies. Moreover, evaluation of invivo x-ray scans revealed that Osteogrow-C with TCP ceramics induced the most rapid defect bridging. On the other hand, control implants (ABC/TCP and ABC/biphasic calcium phosphate) promoted limited osteogenesis without defect bridging. The findings of this study suggest that Osteogrow-C is a promising safe therapeutic solution for the treatment of large bone defects, providing relief to millions of patients suffering from this debilitating condition.

SAR Image Colorization with a Deep Learning Model for All-Inclusion Understanding (H)

The procedure for enhancing halftone photographs with color to create eye-catching pigmented pictures is known as image colorization. This involves the application of colors in line with the intended purpose of the image. However, traditional methods of colorization available today often include the need for end user’s intervention in the form providing color points and doodle drawings, as well as use of color images for reference in which case colors are transferred to the target image or using multiple color images to predict what the colored result would look like; however, the majority of colorizes developed tend to produce colorized images that are not realistic in most instances due to unskilled users who attempt to apply the devices, wrong color transfer or limited color image library. In this paper, to overcome these weaknesses, a new method that consists of two modules is proposed to colorize certain regions of an image by fusing semantic segmentation and seamless region filling. In the first module, input image foreground and background regions masks and categories are extracted each extracted region's reference image is taken from a per-sorted color image database using a Mask R-CNN model. Using colorization approaches based on the VGG and U-Net models, respectively, the second module entails coloring the image's backdrop and other areas. Afterward, the final, fully colored image is created by combining the photographs using the poission editing process. The tests show that our method enables not only the proper reference image selection based on the semantic information of various image components, but also incorporation of the colored results to create a believable colored image. Proposing to use different CNNbased models in a combination where each is used according to its own strength, In contrast to other methods now in use, our scheme gives a deeper creative visual impression while avoiding the disadvantage of single-step techniques' failure frequency. Keywords: Mask R-CNN, Input Photograph, Image Sections, Coloured Photographs, Conceptual Data, Visuals, Black and White Images, Semantic Pix elation, Reference Picture, Base Image, Images Stockpile, Outlying Areas, Image Library, Segregation Methods, Doodle, Conventional Neural Network Based Designs, U-Net Architecture, One-shot Approach, Strategy Present in This Manuscript, Zonal Regions, Principal Zones, Large Image, Color space, Competitive Half toning Algorithm, Zonal Context, Focus Area, Artistic Incorporation, Partially Automatic Process, Cutting Out Section of the Picture, Instance Level Segmentation

Correlation of age to classification of vertical relationship of maxillary sinus and maxillary first molar root by cone-beam computed tomography: a cross-sectional study

Objectives: The maxillary first molar has a close relationship with the base of the maxillary sinus floor. Cone-beam Computed Tomography (CBCT) provides coronal, sagittal, occlusal, and 3D sectional images of maxillofacial structures without causing distortion. Thus, CBCT allows for a comprehensive analysis of the position of the maxillary first molar about the maxillary sinus. This study aims to determine the correlation between age and the classification of vertical relationship between the maxillary sinus and the roots of the maxillary first molar using CBCT. Materials and Methods: The research design was the analytical observational research used a cross-sectional design. The study population includes all CBCT radiographs from patients aged 20-50 years who used CBCT at RSGMP Universitas Jenderal Achmad Yani. The total sampling technique was used to include all CBCT radiograph data comforms to the inclusion and exclusion criteria. Results: The study resulted in 60 CBCT radiographs, with 54 data for the right maxillary first molar and 49 data for the left maxillary first molar. Data analysis using Spearman correlation test showed r = -0.191 with a p-value of 0.166 for the right maxillary first molar and r = -0.167 with a p-value of 0.252 for the left maxillary first molar. Conclusion: There was no correlation between age and the classification of vertical relationship between the maxillary sinus and the maxillary first molar tooth root (p &gt; 0.05). This is because the volume of the maxillary sinus decreases with age, leading to an increased distance between the maxillary sinus and the tooth roots.

Intuitionistic Fuzzy Set Guided Fast Fusion Transformer for Multi-Polarized Petrographic Image of Rock Thin Sections

The fusion of multi-polarized petrographic images of rock thin sections involves the fusion of feature information from microscopic images of rock thin sections illuminated under both plane-polarized and orthogonal-polarized light. During the fusion process of rock thin section images, the inherent high resolution and abundant feature information of the images pose substantial challenges in terms of computational complexity when dealing with massive datasets. In engineering applications, to ensure the quality of image fusion while meeting the practical requirements for high-speed processing, this paper proposes a novel fast fusion Transformer. The model leverages a soft matching algorithm based on intuitionistic fuzzy sets to merge redundant tokens, effectively mitigating the negative effects of asymmetric dependencies between tokens. The newly generated artificial tokens serve as brokers for the Query (Q), forming a novel lightweight fusion strategy. Both subjective visual observations and quantitative analyses demonstrate that the Transformer proposed in this paper is comparable to existing fusion methods in terms of performance while achieving a notable enhancement in its inference efficiency. This is made possible by the attention paradigm, which is equivalent to a generalized form of linear attention, and the newly designed loss function. The model has been experimented on with multiple datasets of different rock types and has exhibited robust generalization capabilities. It provides potential for future research in diverse geological conditions and broader application scenarios.

Development of an H&E on-block staining technique for collagen detection in cryo-fluorescence tomography imaging of frozen breast tissue samples.

Hematoxylin and eosin (H&E) staining is widely considered to be the gold-standard diagnostic tool for histopathology evaluation. However, the fatty nature of some tissue types, such as breast tissue, presents challenges with cryo-sectioning, often resulting in artifacts that can make histopathologic interpretation and correlation with other imaging modalities virtually impossible. We present an optimized on-block H&E staining technique that improves contrast for identifying collagenous stroma during cryo-fluorescence tomography (CFT) sectioning. In this prospective study, we embedded four breast specimens with confirmed ligaments from a bilateral mastopexy in an optimal cutting temperature block. Two of the samples were processed on a CFT imager and stained with our on-block staining protocol. In this protocol, hematoxylin was applied to the block face before being washed with deionized water. Eosin was then applied and washed with 95% ethanol. We then applied mounting medium and acquired images with a stereo-dissecting microscope and camera. Prior to staining, GFP fluorescence and white-light images were acquired with the CFT system to serve as a validation metric. The other two samples were sectioned on a standard cryostat and stained according to gold-standard H&E protocol. The resulting microscope slides were imaged with a digital slide scanner and viewed with Leica Imagescope software. An experienced pathologist evaluated both sets of images for qualitative comparisons. Pathologist evaluation confirmed that striations from on-block staining were qualitatively comparable with collagen tracks identified in gold-standard histology images. Furthermore, GFP images captured collagen autofluorescence, which aligned with the same structures identified by our on-block staining protocol. Our on-block staining technique shows comparable visualization of collagenous structures at the mesoscopic level for fresh breast tissue samples. This technique improves tissue contrast and region of interest selection for histology during CFT imaging for analysis of the stromal architecture of the breast.

Multiphoton Microscopy Assessment of Healing From Tendon Laceration and Microthermal Coagula in a Rat Model.

To study the healing response of rat Achilles tendon when lacerated or treated with intense therapeutic ultrasound (ITU) via utilization of multiphoton microscopy (MPM) imaging and histology. The right Achilles tendon of each Sprague Dawley rat within a cohort was partially lacerated. 1 to 2 days post-surgery, each rat received ITU treatment of the Achilles tendon on either the right or left leg. Rats were euthanized in groups at 1, 3, 7, 14, or 28 days posttreatment and their tendons were explanted, formalin fixed, paraffin embedded, sectioned, and placed on slides for imaging. Slides from each time point were imaged using a laboratory built MPM with a 780 nm Ti:Sapphire laser. The resulting second harmonic generation (SHG) and two-photon excited fluorescence (2PEF) signals were captured, assessed, and compared to brightfield microscopy images of the same section subsequently stained with hematoxylin and eosin. At early timepoints, 2PEF images show the presence of red blood cells, infiltration of inflammatory cells and formation of a fibrin clot at laceration sites, and attraction of fibroblasts to ITU coagula. SHG images indicate an absence of organized collagen in both types of lesions. At later timepoints, new organized collagen can be seen at the laceration sites, and the concentration of inflammatory cells has noticeably decreased. Automated detection of red blood cells and infiltrative cells, as well as analysis of SHG signal intensity and homogeneity was performed at laceration locations. Results show that all quantities except SHG signal intensity approach normal values by day 28. Thus, combined analysis of 2PEF and SHG images elucidates tendon healing processes that align with and complement histological findings. These results indicate that multiphoton imaging can effectively visualize the healing response to mechanical (laceration) and thermal (ITU) injury, including the organization of new collagen which is more difficult to visualize with histology.

Automatic stitching of panoramas for geological images of polished sections

Abstract. This paper addresses the challenge of constructing panoramic images from a set of overlapping microscopic images of geological polished sections.This task is valuable both for automatic mineral detection and for the creating of advanced virtual educational collections. We present LumenStone P1, a new dataset of polished section images for panorama stitching, along with an automatic method for constructing panoramas of these images. Comparisons with existing approaches, including commercial ones, demonstrate that our method is comparable in stitching quality. The numerical value of panorama stitching quality is 0.33 by RMSE for our dataset. The results demonstrate the effectiveness of the proposed method and its practical applicability for the preparation of series of images and further computer analysis of panoramic geological images of polished sections. The LumenStone P1 dataset intended for panorama creation, as well as the source code of the proposed method, are publicly available.