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Mahong4d - The volumetric representation enables a more detailed view of a non-inflamed cholesteatoma (Figure 4). The dynamic mOCT empowers the visualization of specific and characteristic cellular and subcellular structures in the 3D cross-sectional images (Figure 4a). The cholesteatoma with its keratin-accumulated cells appears blue in the dynamic mOCT image, indicating that there is no intracellular motion detectable, which is in line with the presence of large amounts of aggregated keratin, absent or sparse organelles, and the absence of a nucleus. The top view (Figure 4b) clearly shows the irregular structure of the flat keratinized cells from above, which appear as bluish areas. Besides the cellular structures, small green spots are visible singularly and accumulated.

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Kohlfaerber, T.; Pieper, M.; Münter, M.; Holzhausen, C.; Ahrens, M.; Idel, C.; Bruchhage, K.-L.; Leichtle, A.; König, P.; Hüttmann, G.; et al. Dynamic microscopic optical coherence tomography to visualize the morphological and functional micro-anatomy of the airways. Biomed. Opt. Express 2022, 13, 3211–3223. [Google Scholar] [CrossRef]

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Kohlfaerber, T.; Pieper, M.; Münter, Mahong4d M.; Holzhausen, C.; Ahrens, M.; Idel, C.; Bruchhage, K.-L.; Leichtle, A.; König, P.; Hüttmann, G.; et al. Dynamic microscopic optical coherence tomography to visualize the morphological and functional micro-anatomy of the airways. Biomed. Opt. Express 2022, 13, 3211–3223. [Google Scholar] [CrossRef]

The dynamic mOCT volumes were acquired and processed as described in Kohlfaerber et al. [20]. A dynamic mOCT volume consisting of 512 × 512 × 1024 pixels covering a lateral FOV (field of view) of 0.5 × 0.5 mm

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Si, Y.; Chen, Y.B.; Chen, S.J.; Zheng, Y.Q.; Liu, X.; Liu, Y.; Jiang, H.L.; Xu, G.; Li, Z.H.; Huang, Q.H.; et al. TLR4 drives the pathogenesis of acquired cholesteatoma by promoting local inflammation and bone destruction. Sci. Rep. 2015, 5, 16683. [Google Scholar] [CrossRef][Green Version]

Compared to classical OCT, dynamic mOCT provides a much higher information content, which is essential for a reliable diagnosis. While histology also provides this information, it is laborious and time-consuming because the tissue must be sectioned and stained before being observed under a microscope; dynamic mOCT provides dynamic optical sections within a little more than a second. In addition, the tissue does not need to be stained to achieve sufficient contrast to evaluate tissue morphology. Therefore, dynamic mOCT holds the potential to be used to quickly assess excised tissue in the operating theatre without the need for frozen sections.

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Taking a closer look at the connective tissue, it appears as a cell-rich layer (Figure 5d). In addition to the immune cells, which appear as green dots, numerous different cell types of the connective tissue, most likely, fibroblasts, macrophages, and mast cells can be seen.

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PDF) Rice Os9BGlu31 Is a Transglucosidase with the Capacity to Equilibrate Phenylpropanoid, Flavonoid, and Phytohormone Glycoconjugates Figure 2. Comparison of conventional and dynamic mOCT in tissue samples extracted from the middle ear from a patient suffering from chronic otitis media (a,c) and a patient suffering from cholesteatoma (b,d). The different cells and tissue types are challenging to differentiate in the traditional mOCT images of otitis media mesotympanalis (a) and cholesteatoma (b). The dynamic mOCT empowers a significant increase in contrast, so that even individual cell types and tissue regions in the otitis media (c) as well as in the cholesteatoma (d) are visible.

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The identification of the entire cholesteatoma and its complete and residue-free removal poses a significant challenge, as it is crucial to avoid subsequent revision surgeries. So far, there are only a few imaging approaches for the localization of cholesteatoma in situ that go beyond the use of conventional surgical imaging methods.

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All articles published by MDPI are made immediately available worldwide under an open access license. No special permission is required to reuse all or part of the article published by MDPI, including figures and tables. For articles published under an open access Creative Common CC BY license, any part of the article may be reused without permission provided that the original article is clearly cited. For more information, please refer to papers represent the most advanced research with significant potential for high impact in the field. A Feature Paper should be a substantial original Article that involves several techniques or approaches, provides an outlook for future research directions and describes possible research applications.

We were able to demonstrate a marker-free contrast mechanism in the middle ear based on cellular metabolism by exploiting temporal fluctuations of the interferometric signals of dynamic mOCT. These data demonstrate the usefulness of dynamic mOCT in assessing different pathologies of middle ear diseases. Moreover, it opens up an outstanding possibility to study various processes in the middle ear, such as the movement of immune cells, which goes beyond pure morphology such as 3D real-time histology. The technology is promising, supportive of imaging modality, and can potentially increase intra-operative precision during and after otitis media surgery in the future.

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PDF) Scalable production of ultrafine polyaniline fibres for tactile organic electrochemical transistors Leichtle, A.; Klenke, C.; Ebmeyer, J.; Daerr, M.; Bruchhage, K.-L.; Hoffmann, A.S.; Ryan, A.F.; Wollenberg, B.; Sudhoff, H. NOD-Like Receptor Signaling in Cholesteatoma. BioMed Res. Int. 2015, 2015, 408169. [Google Scholar] [CrossRef] [PubMed][Green Version]

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Figure 4. Distinct first-time 3D image of cholesteatoma using dmOCT (a). En face representation (b) shows visualized surface structures with different cells and tissue types of the matrix and keratin layers. The B-scan (c) displays this characteristic picture of a non-inflamed cholesteatoma, confirming cell structures with minimal signal fluctuations. H&E-stained histological section of the cholesteatoma (d). An RGB image depicts the evaluated activity of the cellular scattering structures (red—fast motion (5–25 Hz), green—medium movements (0.5–5 Hz), and blue—slow motion frequencies (0–0.5 Hz)).

Mahong4D Situs Game Mahjong Slot amp Mahong Toto Online © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license A.; Penxova, Z.; Kempin, T.; Leffers, D.; Ahrens, M.; König, P.; Brinkmann, R.; Hüttmann, G.; Bruchhage, K.-L.; Schulz-Hildebrandt, H. Dynamic Microscopic Optical Coherence Tomography as a New Diagnostic Tool for Otitis Media.

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Pieper, M.; Schulz-Hildebrandt, H.; Mall, M.A.; Hüttmann, G.; König, P. Intravital microscopic optical coherence tomography imaging to assess mucus-mobilizing interventions for muco-obstructive lung disease in mice. Am. J. Physiol. Lung Cell. Mol. Physiol. 2020, 318, L518–L524. [Google Scholar] [CrossRef]

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Münter, M.; vom Endt, M.; Pieper, M.; Casper, M.; Ahrens, M.; Kohlfaerber, T.; Rahmanzadeh, R.; König, P.; Hüttmann, G.; Schulz-Hildebrandt, H. Dynamic contrast in scanning microscopic OCT. Opt. Lett. 2020, 45, 4766–4769. [Google Scholar] [CrossRef] [PubMed]

10, no. 6: 685. that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

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Core Oxidized Amorphous Cobalt Phosphide Nanostructures: An Advanced and Highly Efficient Oxygen Evolution Catalyst Immediately after image acquisition, the middle ear tissue samples were fixed and embedded in 4% paraformaldehyde, washed in PBS, and cryoprotected using 20% sucrose in PBS. Tissue was snap frozen in liquid nitrogen and stored at −20 °C to ensure tissue preservation until further processing. Later, 10 µm thick cryo-sections were cut and mounted on slides. The sections were stained with hematoxylin–eosin (H&E) for microscopical morphological tissue inspection.

Mitochondrial dysfunction induces ALK5 SMAD2 mediated hypovascularization and arteriovenous malformations in mouse retinas Jung, J.Y.; Lee, D.H.; Wang, E.W.; Nason, R.; Sinnwell, T.M.; Vogel, J.P.; Chole, R.A. P. aeruginosa infection increases morbidity in experimental cholesteatomas. Laryngoscope 2011, 121, 2449–2454. [Google Scholar] [CrossRef] [PubMed][Green Version]

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Early, S.; Saad, M.A.; Mallidi, S.; Mansour, A.; Seist, R.; Hasan, T.; Stankovic, K.M. A Fluorescent Photoimmunoconjugate for Imaging of Cholesteatoma. Sci. Rep. 2022, 12, 19905. [Google Scholar] [CrossRef]

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Musial, G.; Kohlfaerber, T.; Ahrens, M.; Schulz-Hildebrandt, H.; Steven, P.; Hüttmann, G. Dynamic Contrast Microscopic Optical Coherence Tomography As a Novel Method for Assessing Corneal Epithelium During Exposure to Benzalkonium Chloride. Transl. Vis. Sci. Technol. 2022, 11, 28. [Google Scholar] [CrossRef]

Recreate a Piece of Art in Cinema 4D Mittal, R.; Parrish, J.M.; Soni, M.; Mittal, J.; Mathee, K. Microbial otitis media: Recent advancements in treatment, current challenges and opportunities. J. Med. Microbiol. 2018, 67, 1417–1425. [Google Scholar] [CrossRef]

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Schulz-Hildebrandt, H.; Pieper, M.; Stehmar, C.; Ahrens, M.; Idel, C.; Wollenberg, B.; König, P.; Hüttmann, G. Novel endoscope with increased depth of field for imaging human nasal tissue by microscopic optical coherence tomography. Biomed. Opt. Express 2018, 9, 636–647. [Google Scholar] [CrossRef]

As shown in Figure 2, we can distinguish between the tissue, which was taken out during surgery from a patient suffering from chronic otitis media (Figure 2a) and from a patient suffering from cholesteatoma (Figure 2b).

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With dynamic mOCT, we so far only measured excised tissue, which is susceptible to potential changes in intracellular motion due to a decrease of the metabolism. However, in both longitudinal measurements and volumetric scans, we did not observe any alterations in coloration despite measurement times of up to 30 min. We therefore expect to be able to achieve similar results in vivo.

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Spatial distribution of assessment indicators for hazard of geological... In addition, dynamic mOCT can also be used to observe cell migration. In contrast to other imaging techniques able to visualize the cellular movement, such as fluorescence or multiphoton microscopy, dynamic mOCT does not need extra staining, as is the case for fluorescence microscopy. Furthermore, since dynamic mOCT uses low-intensity near-infrared light, the potential danger of tissue destruction is vastly reduced compared to autofluorescence-based multiphoton microscopy [38,39], which uses high-energy pulses to achieve the two-photon effect necessary for imaging.

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The work was supported by the German Ministry for Education and Research (BMBF) 13N15846 (R.B.) and 13N15847 (A.L.).

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Mahjong 4 Friends on the App Store Thereby, otitis media is characterized by a largely homogeneous scattering on mOCT images (left part of the panel). In this context, using mOCT, structural differences or individual cells are difficult to observe in chronic otitis media (Figure 2a), even though in cholesteatoma at least some individual cells can be distinguished in the extracellular matrix. However, the specific cells and tissue types between the tissue are challenging to differentiate and often impossible to see in the mOCT images. With the dynamic mOCT (right part of the panel), based on the evaluation of intracellular movements, it is possible to increase the contrast significantly.

Schürmann, M.; Oppel, F.; Shao, S.; Volland-Thurn, V.; Kaltschmidt, C.; Kaltschmidt, B.; Scholtz, L.-U.; Sudhoff, H. Chronic inflammation of middle ear cholesteatoma promotes its recurrence via a paracrine mechanism. Cell Commun. Signal. 2021, 19, 25. [Google Scholar] [CrossRef]

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Yung, M.; Tono, T.; Olszewska, E.; Yamamoto, Y.; Sudhoff, H.; Sakagami, M.; Mulder, J.; Kojima, H.; İncesulu, A.; Trabalzini, F.; et al. EAONO/JOS Joint Consensus Statements on the Definitions, Classification and Staging of Middle Ear Cholesteatoma. J. Int. Adv. Otol. 2017, 13, 1–8. [Google Scholar] [CrossRef] [PubMed]

FUNDC1 dependent mitochondria associated endoplasmic reticulum membranes are involved in angiogenesis and neoangiogenesis Fujimoto, J.G.; Pitris, C.; Boppart, S.A.; Brezinski, M.E. Optical coherence tomography: An emerging technology for biomedical imaging and optical biopsy. Neoplasia 2000, 2, 9–25. [Google Scholar] [CrossRef][Green Version]

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Thùng Khẩu trang 4D Yoki hàng 4 lớp công ty thùng khẩu trang kf94 50 hộp/30 túi The extracted human tissue samples were stored immediately in 0.9% NaCl solution and promptly investigated. The samples were placed in a sample holder and sealed with a cover glass before being placed under the dynamic mOCT. The ethics committee of the University of Lübeck approved all protocols (22-041). All clinical examinations were performed according to the highest ethical standards and principles of the Declaration of Helsinki (1964).

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Using dynamic mOCT for imaging an epitympanic polyp of a clinically diagnosed cholesteatoma provides a similar level of detail compared to the histological image (Figure 5). From the acquired 3D volume (Figure 5a), four en face planes are presented (Figure 5b–e). The upper cell layer consists of epithelial cells, which, in contrast to the investigated cholesteatoma in Figure 3, still have intracellular motion indicating cell metabolism (Figure 5b). While the peri-matrix area is depicted in the green channel, the matrix is visible in the blue channel showing little scatterer movements. In a direct comparison of the dynamic mOCT scan with histology of the same tissue (Figure 5g), the matrix (upper level including stratified squamous epithelium with keratinization) and the peri-matrix (lower level including connective tissue and immune cells), can be identified, as we know from cholesteatoma.

Leichtle, A.; Leffers, D.; Daerr, M.G.; Draf, C.; Kurabi, A.; Ryan, A.F.; Rupp, J.; Bruchhage, K.-L. Immunmodulation im Cholesteatom. Laryngorhinootologie 2022, 101, 310–319. [Google Scholar] [CrossRef]

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Mishiro, Y.; Sakagami, M.; Kitahara, T.; Kondoh, K.; Okumura, S. The investigation of the recurrence rate of cholesteatoma using Kaplan-Meier survival analysis. Otol. Neurotol. 2008, 29, 803–806. [Google Scholar] [CrossRef]

Figure 5. 3D image of an epitympanic polyp and visualization of cell migration using dmOCT (a). En face representations of the polyp (b–e) show visualized surface structures with different cells and tissue types of the matrix, peri-matrix, and keratin layers. In the time-series of B-scans (f), live cell movement was observed. H&E-stained histological section of the polyp (g). An RGB image depicts the evaluated cellular movement activity (red—fast motion (5–25 Hz), green—medium movements (0.5–5 Hz) and blue—slow motion frequencies (0–0.5 Hz)).

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Leichtle A, Penxova Z, Kempin T, Leffers D, Ahrens M, König P, Brinkmann R, Hüttmann G, Bruchhage K-L, Schulz-Hildebrandt H. Dynamic Microscopic Optical Coherence Tomography as a New Diagnostic Tool for Otitis Media.

In Figure 5b, bright green cells are visible. Based on the results of our previous work by Kohlfaerber et al. [20] and the fact that they are present in all layers, we can assume that these are immune cells that have migrated into the epithelium. The following en face plane (Figure 5c) cuts through the epithelium, the lamia propria, as well as the connective tissue. The fibrous structure in the lamia propria appears blue, indicating little activity of this tissue.

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4 PLAYER MAHJONG Free Mahjong Online Games This is even more evident in the cross-sectional image of the epitympanic polyp (Figure 5f), showing the epithelium displays stratified squamous epithelium (matrix) in the upper part of the image. Here, the dynamic mOCT B-scan of the epitympanic polyp shows distinct tissue layers of keratin, matrix, and peri-matrix, equally to our corresponding histological image (Figure 5g). Although the polyp looks macroscopically more like inflamed mucosal middle ear tissue (Figure 5b) than a non-inflamed cholesteatoma (Figure 5c), we do see by dynamic mOCT and by histology characteristic images of both inflammation and cholesteatoma, what we know from established histological pictures of inflammatory attached triggered cholesteatomas.

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Microscopically and macroscopically, the tissue looks like a regular inflamed mucosal polyp of otitis media mesotympanalis. Using 3D imaging with dynamic mOCT, it seems feasible to discriminate the epitympanic polyp from classical inflamed tissue.

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Brazaletes 3/4″ (pulseras de tyvek) Qin, J.; An, L. Optical Coherence Tomography for Ophthalmology Imaging. Adv. Exp. Med. Biol. 2021, 3233, 197–216. [Google Scholar] [CrossRef] [PubMed]

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PDF) Cost Effective Trap qPCR Approach to Evaluate Telomerase Activity: an Important Tool for Aging, Cancer, and Chronic Disease Research Interestingly, the polyp seems to trigger the cholesteatoma growth via inflammation, which can be followed by dmOCT as we can see in the surrounding tissue multiple migrated cells, which most likely are immune cells that have migrated into the epithelium, accordingly to our previous results [20]. We underline this theory since we discovered a live cell movement in the peri-matrix, Furthermore, this was assessed by the clinical histopathological evaluation, revealing stacks of keratin and epithelial cells decomposed with immune and inflammatory cells corresponding to the picture of a medium to high-grade inflammatory cholesteatoma. Several studies associate the interaction of the immune system with the cholesteatoma [6,25,26,27,28,29,36] and even specifically target innate immunoregulatory events [37]. Unfortunately, to date, there are no markers for OCT, which are able to label specific immune cells. For evaluating and detecting specific otitis media and cholesteatoma markers in dynamic mOCT compared to H&E staining, further studies with a higher number of tissue samples and quantitative analysis need to be performed and are under ongoing investigation.

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Akt1 dependent expression of angiopoietin 1 and 2 in vascular smooth muscle cells leads to vascular stabilization Apelian, C.; Harms, F.; Thouvenin, O.; Boccara, A.C. Dynamic full field optical coherence tomography: Subcellular metabolic contrast revealed in tissues by interferometric signals temporal analysis. Biomed. Opt. Express 2016, 7, 1511–1524. [Google Scholar] [CrossRef][Green Version]

Arguedas, A.; Kvaerner, K.; Liese, J.; Schilder, A.G.M.; Pelton, S.I. Otitis media across nine countries: Disease burden and management. Int. J. Pediatr. Otorhinolaryngol. 2010, 74, 1419–1424. [Google Scholar] [CrossRef] [PubMed]

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Aumann, S.; Donner, S.; Fischer, J.; Müller, F. High Resolution Imaging in Microscopy and Ophthalmology: New Frontiers in Biomedical Optics; Springer: Cham, Switzerland, 2019; ISBN 9783030166373. [Google Scholar]

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Mahongtoto Situs Judi Togel Casino Online Slot Online . 2023; 10(6):685. Anke, Zuzana Penxova, Thorge Kempin, David Leffers, Martin Ahrens, Peter König, Ralf Brinkmann, Gereon Hüttmann, Karl-Ludwig Bruchhage, and Hinnerk Schulz-Hildebrandt. 2023. "Dynamic Microscopic Optical Coherence Tomography as a New Diagnostic Tool for Otitis Media"

Using our new dynamic mOCT imaging modality, we demonstrated 3D marker-free images of a characteristic less inflamed cholesteatoma pathology. The images not only showed in the cholesteatoma keratin accumulating cells in white or blue but allowed us to evaluate overlying layers of keratin in an entire texture en face and in B-scan as a mesh, showing little detectable intracellular scatterer movement with less nuclei or organelles. These findings are supported by our histologic studies and by others [25].

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Figure 1. Schematic drawing of the mOCT setup showing the various components used for scanning and data acquisition. The setup includes a 50/50 fiber coupler (FC), collimators (C1/C2), galvanometer mirror scanners (XY scanner), beam expander lenses (L1, L2), a microscope objective (L3), dispersion compensation (DC), a retroreflector (RR), a data acquisition device (DAQ), and a computer (PC) for scanning control and data processing.

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PDF) The final piece of the Triangle of U: Evolution of the tetraploid Brassica carinata genome AWMF. 017-074l_S2k_Chronisch-Mesotympanale-Otitis-Media_2020-11_01. Available online: (accessed on 15 March 2023).

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Jual Perjuangan! 71jt Cair dari Bisnis Ini. Tag: Www Savegrab Online Pengeluaran Mahong 4D Html Harga Terbaik & Termurah September 2023 Editor’s Choice articles are based on recommendations by the scientific editors of MDPI journals from around the world. Editors select a small number of articles recently published in the journal that they believe will be particularly interesting to readers, or important in the respective research area. The aim is to provide a snapshot of some of the most exciting work published in the various research areas of the journal.

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Schilder, A.G.M.; Chonmaitree, T.; Cripps, A.W.; Rosenfeld, R.M.; Casselbrant, M.L.; Haggard, M.P.; Venekamp, R.P. Otitis media. Nat. Rev. Dis. Prim. 2016, 2, 16063. [Google Scholar] [CrossRef] [PubMed]

The study was conducted in accordance with the Declaration of Helsinki and approved by the Ethics Committee of University of Lübeck (22-041).

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The authors gratefully acknowledge Kathy Budler, Jasmin Koritke, and Petra Lau from the Institute of Anatomy at the University of Lübeck for their assistance and training with the H&E slides, histology, and microscopy.

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How to Play Mahjong Solitaire For Beginners: Rules & Strategies Before dynamic mOCT can be used in vivo, technical solutions and advanced image registration methods must be developed to reduce susceptibility to global motion. One promising approach is to perform dynamic mOCT calculations on volumes instead of single B-scans since registration can be carried out in all three dimensions [21]. After solving the current challenges of combining dynamic mOCT with mOCT endoscopy [40], the technology has a high potential for intra-operative assessment of the middle ear tissue during surgery.

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Oil Filter BMW 1 3 5 7 Series X1 X3 2.0 3.0 Diesel MAHLE OX361/4D 4009026511480 Early et al., investigated photosensitizer immunoconjugate (PIC) cetuximab benzoporphyrin derivative (Cet-BPD) for intraoperative localization of human cholesteatoma tissue and examined its potential in an animal study [30]. Although the results are promising, the ototoxicity of Cet-BPD in humans is not clear, and side effects such as skin rash are common. Another approach pursued by various research groups is the use of classical OCT. They were able to show that clinically distinct cholesteatomas can be identified based on the appearance of a greater signal intensity of the cholestatoma compared with the lower grayscale values of the mucosa [31]. They could only show that clinically distinct cholestatomas can be identified based on the appearance of a larger signal intensity of the cholestatoma compared with the lower grayscale values of the mucosa. Differentiation of moderately inflamed tissue from normal mucosa was not possible, and the resolution was not high enough to see individual cells. In contrast, dynamic OCT does not require external markers and uses harmless near-infrared light and intracellular motion to provide virtual coloration.

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In industrialized nations, otitis media (OM) is one of the diseases with a significantly high socio-economic burden on the healthcare system [1,2] due to an increased number of medical examinations, antibiotic prescriptions, and surgeries [3]. In developing countries, OM is a considerable health issue with a high infant mortality [1]. Chronic otitis media (COM) significantly affects the patient’s quality of life through ear discharge, hearing loss, and speech development delays and thus can result in complete deafness and dizziness [4,5]. This is even more evident in the course of a particularly aggressive form of COM, which is known as chronic otitis media epitympanalis. Untreated, it leads in most cases to a cholesteatoma. The cholesteatoma evades antibiotic therapy and behaves like a malignant tumor through progressive inflammatory destructive processes of the middle ear and the surrounding structures [6,7,8].

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Diversity and evolution of nitric oxide reduction Conceptualization, A.L., K.-L.B. and H.S.-H.; methodology, A.L., H.S.-H., G.H., T.K., P.K. and R.B.; software, M.A. and T.K.; validation, A.L., H.S.-H., T.K., Z.P. and P.K.; formal analysis, D.L.; investigation, A.L., Z.P., T.K., D.L. and H.S.-H.; resources, A.L., R.B., G.H., P.K. and K.-L.B.; writing—original draft preparation, A.L., H.S.-H. and K.-L.B.; writing—review and editing A.L., D.L., P.K., R.B., G.H., K.-L.B. and H.S.-H.; visualization, A.L. and H.S.-H.; supervision, A.L. and H.S.-H.; funding acquisition, A.L., K.-L.B. and R.B. All authors have read and agreed to the published version of the manuscript.

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Monroy, G.L.; Won, J.; Shi, J.; Hill, M.C.; Porter, R.G.; Novak, M.A.; Hong, W.; Khampang, P.; Kerschner, J.E.; Spillman, D.R.; et al. Automated classification of otitis media with OCT: Augmenting pediatric image datasets with gold-standard animal model data. Biomed. Opt. Express 2022, 13, 3601–3614. [Google Scholar] [CrossRef] [PubMed]

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Informed consent was obtained from all subjects involved in the study.

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Moving the en face plane further away from the epithelium, we can see that the connective tissue becomes increasingly difficult to discriminate (Figure 5e). However, despite being far from the optical focus, dynamic mOCT still allows us to obtain information such as the presence of inflammatory cells.

Despite intensive research, the only definitive therapy for progressive COM and cholesteatoma for decades has been surgical treatment with a complete removal of the diseased focus [9,10].

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Diverse physiological and pathological tissue samples of the middle ear were obtained from 12 patients who had undergone otologic surgery at the Otorhinolaryngology department of the University Medical Center Schleswig-Holstein in Lübeck (Germany). Eight patients were diagnosed clinically and radiologically with COM or cholesteatoma. Moreover, tissue of these pathologies was sent and evaluated by our pathologists according to the standard procedure during ear surgery. Healthy middle ear tissue from four patients was collected from a tympanoplasty and three cochlear implantations.

Structural analysis of rice Os4BGlu18 monolignol β glucosidase Here we demonstrate for the first time the use of marker-free virtual staining by dmOCT to observe multiple middle-ear diseases in their diverse pathologies.

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Wan, B.; Ganier, C.; Du-Harpur, X.; Harun, N.; Watt, F.M.; Patalay, R.; Lynch, M.D. Applications and future directions for optical coherence tomography in dermatology. Br. J. Dermatol. 2021, 184, 1014–1022. [Google Scholar] [CrossRef] [PubMed]

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Klenke, C.; Janowski, S.; Borck, D.; Widera, D.; Ebmeyer, J.; Kalinowski, J.; Leichtle, A.; Hofestädt, R.; Upile, T.; Kaltschmidt, C.; et al. Identification of novel cholesteatoma-related gene expression signatures using full-genome microarrays. PLoS ONE 2012, 7, e52718. [Google Scholar] [CrossRef] [PubMed][Green Version]

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Kretschmer, S.; Pieper, M.; Hüttmann, G.; Bölke, T.; Wollenberg, B.; Marsh, L.M.; Garn, H.; König, P. Autofluorescence multiphoton microscopy for visualization of tissue morphology and cellular dynamics in murine and human airways. Lab. Investig. 2016, 96, 918–931. [Google Scholar] [CrossRef][Green Version]

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Germain, R.N.; Miller, M.J.; Dustin, M.L.; Nussenzweig, M.C. Dynamic imaging of the immune system: Progress, pitfalls and promise. Nat. Rev. Immunol. 2006, 6, 497–507. [Google Scholar] [CrossRef] [PubMed]

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Thouvenin, O.; Boccara, C.; Fink, M.; Sahel, J.; Pâques, M.; Grieve, K. Cell Motility as Contrast Agent in Retinal Explant Imaging With Full-Field Optical Coherence Tomography. Investig. Ophthalmol. Vis. Sci. 2017, 58, 4605–4615. [Google Scholar] [CrossRef][Green Version]

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Identification of Active Sites on High Performance Pt/Al2O3 Catalyst for Cryogenic CO Oxidation Imaging via dynamic mOCT allows high-resolution imaging of living tissues to be obtained. This allows the differentiation of various middle ear pathologies that differ morphologically (Figure 3a), like an epitympanic polyp (Figure 3b), a cholesteatoma (Figure 3c), and a normal middle ear mucosa (Figure 3d).

Ligand Control of E/Z Selectivity in Nickel Catalyzed Transfer Hydrogenative Alkyne Semireduction While the histological image provides only static information, such as cell morphology, dynamic mOCT can also visualize long-timescale dynamics like cell migration. To demonstrate this, we acquired cross-sectional images at the same position over 10 min every 10 s (Figure 5f), following a modified imaging protocol of our previous work [20], which allowed the evaluation of the time series to track individual cells over time. In the supplemented video to Figure 5f, when zoomed in, it can be observed that a single cell moves upward and eventually disappears while the surrounding tissue remains unchanged.

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Haile, L.M.; Kamenov, K.; Briant, P.S.; Orji, A.U.; Steinmetz, J.D.; Abdoli, A.; Abdollahi, M.; Abu-Gharbieh, E.; Afshin, A.; Ahmed, H.; et al. Hearing loss prevalence and years lived with disability, 1990–2019: Findings from the Global Burden of Disease Study 2019. Lancet 2021, 397, 996–1009. [Google Scholar] [CrossRef] [PubMed]

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Using our microscopic optical coherence tomography imaging setup in Figure 2, we gained high-resolution images of living tissues, giving a good impression of the general morphology.

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Well known from the microscopic view during surgery, a cholesteatoma may present with white keratin layers surrounded by inflammatory tissue, representing different cells and tissue types of the matrix and peri-matrix. Thereby, the cholesteatoma varies from showing characteristics of minor to high inflammation, which are reflected in the presence or absence of inflammatory cells [25,26,27,28,29].

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Mahongtoto Daftar Login Wap So far, successful surgical removal of the diseased tissue is limited to the anatomical localization, extensiveness, and diffusiveness of the cholesteatoma; thereby, it represents a surgical challenge aggravated by the twisted anatomy of the middle ear and the petrous part of the temporal bone. Consequently, at worse-case scenario, it is necessary to create a radical cavity by removing the auditory ossicles, exposing the facial nerve, and sometimes even removing the inner ear and vestibular structures. This approach necessitates a comprehensive surgical procedure to gain the required overview and control in the middle ear and the petrous part of the temporal bone during surgery, entailing increased morbidity and mortality. In addition, a more prominent inflammatory component is a strong negative predictor [6] and becomes lethal when the cholesteatoma spreads to the brain tissue. Furthermore, a high tendency to recurrence of over 30% with an increased number of unreported cases has been estimated [11]. Additionally, more than 70% of patients require another ear operation within ten years, as a “second-look surgery” in order to exclude recurrences and, if necessary, to rebuild the auditory structures for a significant hearing improvement and a significant better quality of life [6]. A post-surgical lifelong state comprises clinical and radiological follow-ups, hospital stays, and potentially further surgeries. Developing new diagnostic and theragnostic surgical concepts is of urgent relevance concerning the high chronification and recurrence rate, the increased antibiotic resistance, further complication risks, and the enormous costs for the health care system. Thereby, a complete and safe resection of the middle ear pathology can only be guaranteed by the reliable in vivo identification of the destructive processes in distinction to the healthy middle ear mucosa and bone tissue. Nowadays, such distinguishing is predominantly based on the surgeons’ visual perception and experience. So far, supportive diagnostic imaging modalities have been little explored and have not found their way into clinical application. Finding imaging techniques that can identify inflamed/cholesteatoma tissue intraoperatively would present an important tool to support surgeons by identifying tissue that must be removed during surgery.

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Leung, H.M.; Wang, M.L.; Osman, H.; Abouei, E.; MacAulay, C.; Follen, M.; Gardecki, J.A.; Tearney, G.J. Imaging intracellular motion with dynamic micro-optical coherence tomography. Biomed. Opt. Express 2020, 11, 2768–2778. [Google Scholar] [CrossRef]

OCT is based on light scattering by subcellular and sub-resolution structures in biological tissues. Whereas layered structures and borders of tissue compartments with different average scattering can be visualized by OCT, even at high resolution it is difficult to discriminate cellular and subcellular structures. However, a greatly increased cellular contrast was achieved by evaluating the intracellular movement of the different scatterers and displaying the fluctuation frequencies in three frequency bands, where slow movements were coded in blue, medium movements in green, and fast movements in red [20]. Active cells, such as epithelium or inflammatory cells were displayed in the green and red channels. In contrast, inactive tissue structures such as keratinized cells in cholesteatoma, the fibrous structures in the connective tissue, or the extracellular matrix were displayed in blue. Although the biological origin of dynamic mOCT is not entirely understood, in this work it is a reasonable assumption that scattering fluctuations are influenced by cellular metabolism.

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and an axial depth of 890 µm was recorded with an A-scan rate of 100 kHz. A volume was acquired by taking 150 repetitive B-scans (cross-sectional view) at each cross-sectional scan position with a B-scan acquisition rate of 111 Hz (taking 1.35 s), thus the total volume acquisition time for 512 scan lines accounts for 11.52 min. MATLAB (MATLAB R2019b, The MathWorks, Inc., Natick, MA, USA) was used for the spectral processing and dynamic contrast enhancement of the data. The spectrometer’s raw data were linearized, Hann-windowed, and Fourier-transformed to generate the OCT images. Numerical compensation for residual dispersion was performed using a fifth-order polynomial to correct the spectral phase error. The coefficients of the polynomial were determined by optimizing image quality, assessed by the Shannon entropy measurement. Intracellular motion resulting in signal fluctuations was analyzed in a frequency range of up to 55.5 Hz by temporal Fourier transformation and summing up the integral amplitudes in three bands. Each channel was then normalized from 0 to 1. Next, a moving standard deviation image was generated with a window width of 25 images, and the histogram of each channel was matched to this image. The three frequency bands were stacked together into an RGB image where fast fluctuations (4.9–25 Hz) are coded in the red channel, medium motion (0.4–4.9 Hz) in the green channel and slow motion (<0.4 Hz) in the blue channel. The three frequency bands are based on an early work using FF-OCT [23] and matching well-known frequencies of intracellular motion [24]. ImageJ/Fiji (ImageJ-win64)was used for analyzing and visualizing the data.

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In this study, we could illustrate striking differences in dynamic mOCT images in otitis media mesotympanalis, and otitis media epitympanalis (cholesteatoma), including an epitympanic polyp, regarding tissue structures, subcellular structures, and the appearance of individual cells using temporal fluctuations of the light scattering structures. This is a main diagnostic advancement in comparison to studies using conventional/commercial OCT imaging systems in otitis media [32,33,34].

In this context, especially the observed difference between the cholesteatoma and the epitympanic polyp is of great interest since we know these polyps are named “signal polyps” of a cholesteatoma, often hidden behind a defect tympanum. In clinical practice, this belongs to one of the otoscopic signs of a possible cholesteatoma, even if the patient presents without perforation, white keratin mass in the epitympanic part, or other symptoms, such as otorrhea, hearing loss, or dizziness [9,35].

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PDF) All‐in‐One Compression and Encryption Engine Based on Flexible Polyimide Memristor Different tissue areas and cells can be identified in both chronic otitis media (Figure 2c), showing different tissue textures by cell movement depicted in green, yellow, and red and in cholesteatoma (Figure 2d), showing decreased intracellular movement depicted in blue-purple.

Figure 3. Overview of macroscopic tissue samples of the middle ear and their dmOCT image. Exemplary images of extracted tissue from the middle ear (a): epitympanic polyp (b), cholesteatoma (c), and middle ear mucosa (d). The different cells and tissue types can be clearly differentiated in the dmOCT images. The signal fluctuations inside the cells are represented and coded as an RGB image (red—fast motion (5–25 Hz), green—medium movements (0.5–5 Hz), and blue—slow motion frequencies (0–0.5 Hz)).

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While we know that these are particles in the surrounding media and on top of the cholesteatoma, we cannot prove if these are cells, bacteria, or debris due to tissue removal. The B-Scan with lamellar sheets of keratin (Figure 4c) shows low activity with a high structural similarity to the direct comparison with the histological section (Figure 4d).

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Membrane association of importin α facilitates viral entry into salivary gland cells of vector insects Chole, R.A.; Gagnon, P.M.; Vogel, J.P. Inactivation of specific Pseudomonas aeruginosa biofilm factors does not alter virulence in infected cholesteatomas. Otol. Neurotol. 2014, 35, 1585–1591. [Google Scholar] [CrossRef][Green Version]

Djalilian, H.R.; Rubinstein, M.; Wu, E.C.; Naemi, K.; Zardouz, S.; Karimi, K.; Wong, B.J.F. Optical coherence tomography of cholesteatoma. Otol. Neurotol. 2010, 31, 932–935. [Google Scholar] [CrossRef][Green Version]

Optical coherence tomography (OCT) is a non-contact, low coherence infrared light-based scanning technique that images optical structures in biological tissue [12,13]. Like ultrasound, OCT measures the echo time delay in tissues but uses light instead of sound. Since light travels significantly faster, interferometers are used to measure the time delays. It is frequently used in ophthalmology [14] and dermatology [15]. With a spatial resolution of around 10 µm to 30 µm, it is well suited for imaging layer structures like the retina and the skin. Unfortunately, it fails to provide a sufficient resolution for visualizing cellular structures. By extending OCT with broader light sources and high numerical aperture objectives, a resolution down to 1 µm can be reached [16]. While this microscopic OCT (mOCT) provides an excellent resolution to visualize cells, the lack of contrast between cells and surrounding connective tissue inhibits cell phenotyping. Therefore, its use as in situ optical histology is limited. While there are no contrast agents available for mOCT, recently remarkable contrast enhancement was achieved by evaluating signal fluctuations based on the intracellular motion [17] induced by ATP consumption. While it was demonstrated first with full-field OCT systems [17], which are quite challenging to translate to in vivo settings, it has since been shown to be feasible with scanning microscopic OCT systems in different tissue types [16,18,19,20].

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Hypothesis: Otitis media (OM) can be successfully visualized and diagnosed by dynamic microscopic optical coherence tomography (dmOCT). Background: OM is one of the most common infectious diseases and, according to the WHO, one of the leading health problems with high mortality in developing countries. Despite intensive research, the only definitive treatment of therapy-refractory OM for decades has been the surgical removal of inflamed tissue. Thereby, the intra-operative diagnosis is limited to the surgeon’s visual impression. Supportive imaging modalities have been little explored and have not found their way into clinical application. Finding imaging techniques capable of identifying inflamed tissue intraoperatively, therefore, is of significant clinical relevance. Methods: This work investigated a modified version of optical coherence tomography with a microscopic resolution (mOCT) regarding its ability to differentiate between healthy and inflamed tissue. Despite its high resolution, the differentiation of single cells with mOCT is often impossible. A new form of mOCT termed dynamic mOCT (dmOCT) achieves cellular contrast using micro-movements within cells based on their metabolism. It was used in this study to establish correlative measurements with histology. Results: Using dmOCT, images with microscopic resolution were acquired on ex vivo tissue samples of chronic otitis media and cholesteatoma. Imaging with dmOCT allowed the visualization of specific and characteristic cellular and subcellular structures in the cross-sectional images, which can be identified only to a limited extent in native mOCT. Conclusion: We demonstrated for the first time a new marker-free visualization in otitis media based on intracellular motion using dmOCT.

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Lui, C.G.; Kim, W.; Dewey, J.B.; Macías-Escrivá, F.D.; Ratnayake, K.; Oghalai, J.S.; Applegate, B.E. In vivo functional imaging of the human middle ear with a hand-held optical coherence tomography device. Biomed. Opt. Express 2021, 12, 5196–5213. [Google Scholar] [CrossRef]

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Different human tissue samples from the region of interest were surgically obtained and collected to simulate a situation similar to an optical biopsy during middle ear surgery due to progressive otitis media and cholesteatoma. Ex vivo examinations of tissue probes were carried out employing a prototype of a dynamic mOCT device. The acquired and reconstructed images were correlated with standard histological staining of the examined tissue.

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Ahmed, S.; Shapiro, N.L.; Bhattacharyya, N. Incremental health care utilization and costs for acute otitis media in children. Laryngoscope 2014, 124, 301–305. [Google Scholar] [CrossRef] [PubMed]

Münter, M.; Pieper, M.; Kohlfaerber, T.; Bodenstorfer, E.; Ahrens, M.; Winter, C.; Huber, R.; König, P.; Hüttmann, G.; Schulz-Hildebrandt, H. Microscopic optical coherence tomography (mOCT) at 600 kHz for 4D volumetric imaging and dynamic contrast. Biomed. Opt. Express 2021, 12, 6024–6039. [Google Scholar] [CrossRef]

Münter, M.; Pieper, Mahong4d M.; Kohlfaerber, T.; Bodenstorfer, E.; Ahrens, M.; Winter, C.; Huber, R.; König, P.; Hüttmann, G.; Schulz-Hildebrandt, H. Microscopic optical coherence tomography (mOCT) at 600 kHz for 4D volumetric imaging and dynamic contrast. Biomed. Opt. Express 2021, 12, 6024–6039. [Google Scholar] [CrossRef]

Dynamic Microscopic Optical Coherence Tomography as a New Diagnostic Tool for Otitis Media

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