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Lysosomes serve dual antagonistic functions in cells by mediating anabolic growth signaling and the catabolic turnover of macromolecules. How these janus-faced activities are regulated in response to cellular nutrient status is poorly understood. We show here that lysosome morphology and function are reversibly controlled by a nutrient-regulated signaling lipid switch that triggers the conversion between peripheral motile mTOR complex 1 (mTORC1) signaling-active and static mTORC1-inactive degradative lysosomes clustered at the cell center. Starvation-triggered relocalization of phosphatidylinositol 4-phosphate (PI(4)P)-metabolizing enzymes reshapes the lysosomal surface proteome to facilitate lysosomal proteolysis and to repress mTORC1 signaling. Concomitantly, lysosomal phosphatidylinositol 3-phosphate (PI(3)P), which marks motile signaling-active lysosomes in the cell periphery, is erased. Interference with this PI(3)P/PI(4)P lipid switch module impairs the adaptive response of cells to altering nutrient supply. Our data unravel a key function for lysosomal phosphoinositide metabolism in rewiring organellar membrane dynamics in response to cellular nutrient status.
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Lisosomas , Transducción de Señal , Lisosomas/metabolismo , Diana Mecanicista del Complejo 1 de la Rapamicina/metabolismo , Nutrientes , Fenómenos Fisiológicos CelularesRESUMEN
Images document scientific discoveries and are prevalent in modern biomedical research. Microscopy imaging in particular is currently undergoing rapid technological advancements. However, for scientists wishing to publish obtained images and image-analysis results, there are currently no unified guidelines for best practices. Consequently, microscopy images and image data in publications may be unclear or difficult to interpret. Here, we present community-developed checklists for preparing light microscopy images and describing image analyses for publications. These checklists offer authors, readers and publishers key recommendations for image formatting and annotation, color selection, data availability and reporting image-analysis workflows. The goal of our guidelines is to increase the clarity and reproducibility of image figures and thereby to heighten the quality and explanatory power of microscopy data.
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Lista de Verificación , Edición , Reproducibilidad de los Resultados , Procesamiento de Imagen Asistido por Computador , MicroscopíaRESUMEN
Phosphoinositides are amphipathic lipid molecules derived from phosphatidylinositol that represent low abundance components of biological membranes. Rather than serving as mere structural elements of lipid bilayers, they represent molecular switches for a broad range of biological processes, including cell signaling, membrane dynamics and remodeling, and many other functions. Here, we focus on the molecular mechanisms that turn phosphoinositides into molecular switches and how the dysregulation of these processes can lead to disease.
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Enfermedad , Fosfatidilinositoles , Transducción de Señal , Membrana Celular/metabolismo , Fosfatidilinositoles/metabolismo , HumanosRESUMEN
Protein kinase B/Akt regulates cellular metabolism, survival, and proliferation in response to hormones and growth factors. Hyperactivation of Akt is frequently observed in cancer, while Akt inactivation is associated with severe diabetes. Here, we investigated the molecular and cellular mechanisms that maintain Akt activity proportional to the activating stimulus. We show that binding of phosphatidylinositol-3,4,5-trisphosphate (PIP3) or PI(3,4)P2 to the PH domain allosterically activates Akt by promoting high-affinity substrate binding. Conversely, dissociation from PIP3 was rate limiting for Akt dephosphorylation, dependent on the presence of the PH domain. In cells, active Akt associated primarily with cellular membranes. In contrast, a transforming mutation that uncouples kinase activation from PIP3 resulted in the accumulation of hyperphosphorylated, active Akt in the cytosol. Our results suggest that intramolecular allosteric and cellular mechanisms cooperate to restrict Akt activity to cellular membranes, thereby enhancing the fidelity of Akt signaling and the specificity of downstream substrate phosphorylation.
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Membrana Celular/metabolismo , Fosfatidilinositoles/metabolismo , Proteínas Proto-Oncogénicas c-akt/química , Proteínas Proto-Oncogénicas c-akt/metabolismo , Regulación Alostérica , Sitios de Unión , Regulación de la Expresión Génica , Células HeLa , Humanos , Células MCF-7 , Mutación , Fosforilación , Unión Proteica , Proteínas Proto-Oncogénicas c-akt/genética , Especificidad por SustratoRESUMEN
Lysosomes serve as dynamic regulators of cell and organismal physiology by integrating the degradation of macromolecules with receptor and nutrient signaling. Previous studies have established that activation of the transcription factor EB (TFEB) and transcription factor E3 (TFE3) induces the expression of lysosomal genes and proteins in signaling-inactive starved cells, that is, under conditions when activity of the master regulator of nutrient-sensing signaling mechanistic target of rapamycin complex 1 is repressed. How lysosome biogenesis is triggered in signaling-active cells is incompletely understood. Here, we identify a role for calcium release from the lumen of the endoplasmic reticulum in the control of lysosome biogenesis that is independent of mechanistic target of rapamycin complex 1. We show using functional imaging that calcium efflux from endoplasmic reticulum stores induced by inositol triphosphate accumulation upon depletion of inositol polyphosphate-5-phosphatase A, an inositol 5-phosphatase downregulated in cancer and defective in spinocerebellar ataxia, or receptor-mediated phospholipase C activation leads to the induction of lysosome biogenesis. This mechanism involves calcineurin and the nuclear translocation and elevated transcriptional activity of TFEB/TFE3. Our findings reveal a crucial function for inositol polyphosphate-5-phosphatase A-mediated triphosphate hydrolysis in the control of lysosome biogenesis via TFEB/TFE3, thereby contributing to our understanding how cells are able to maintain their lysosome content under conditions of active receptor and nutrient signaling.
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Autofagia , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice , Calcio , Retículo Endoplásmico , Lisosomas , Polifosfatos , Autofagia/fisiología , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Calcineurina/metabolismo , Calcio/metabolismo , Retículo Endoplásmico/metabolismo , Inositol/metabolismo , Lisosomas/metabolismo , Diana Mecanicista del Complejo 1 de la Rapamicina/metabolismo , Polifosfatos/metabolismoRESUMEN
BACKGROUND: Anastomotic leak is one of the most feared complications of colorectal surgery, and probably linked to poor blood supply to the anastomotic site. Several technologies have been described for intraoperative assessment of bowel perfusion. This systematic review and meta-analysis aimed to evaluate the most frequently used bowel perfusion assessment modalities in elective colorectal procedures, and to assess their associated risk of anastomotic leak. Technologies included indocyanine green fluorescence angiography, diffuse reflectance spectroscopy, laser speckle contrast imaging, and hyperspectral imaging. METHODS: The review was preregistered with PROSPERO (CRD42021297299). A comprehensive literature search was performed using Embase, MEDLINE, Cochrane Library, Scopus, and Web of Science. The final search was undertaken on 29 July 2022. Data were extracted by two reviewers and the MINORS criteria were applied to assess the risk of bias. RESULTS: Some 66 eligible studies involving 11 560 participants were included. Indocyanine green fluorescence angiography was most used with 10 789 participants, followed by diffuse reflectance spectroscopy with 321, hyperspectral imaging with 265, and laser speckle contrast imaging with 185. In the meta-analysis, the total pooled effect of an intervention on anastomotic leak was 0.05 (95 per cent c.i. 0.04 to 0.07) in comparison with 0.10 (0.08 to 0.12) without. Use of indocyanine green fluorescence angiography, hyperspectral imaging, or laser speckle contrast imaging was associated with a significant reduction in anastomotic leak. CONCLUSION: Bowel perfusion assessment reduced the incidence of anastomotic leak, with intraoperative indocyanine green fluorescence angiography, hyperspectral imaging, and laser speckle contrast imaging all demonstrating comparable results.
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Fuga Anastomótica , Procedimientos Quirúrgicos del Sistema Digestivo , Humanos , Fuga Anastomótica/etiología , Fuga Anastomótica/prevención & control , Fuga Anastomótica/epidemiología , Verde de Indocianina , Anastomosis Quirúrgica/efectos adversos , Anastomosis Quirúrgica/métodos , Procedimientos Quirúrgicos del Sistema Digestivo/efectos adversos , Procedimientos Quirúrgicos del Sistema Digestivo/métodos , PerfusiónRESUMEN
Motion correction is an essential preprocessing step in functional Magnetic Resonance Imaging (fMRI) of the fetal brain with the aim to remove artifacts caused by fetal movement and maternal breathing and consequently to suppress erroneous signal correlations. Current motion correction approaches for fetal fMRI choose a single 3D volume from a specific acquisition timepoint with least motion artefacts as reference volume, and perform interpolation for the reconstruction of the motion corrected time series. The results can suffer, if no low-motion frame is available, and if reconstruction does not exploit any assumptions about the continuity of the fMRI signal. Here, we propose a novel framework, which estimates a high-resolution reference volume by using outlier-robust motion correction, and by utilizing Huber L2 regularization for intra-stack volumetric reconstruction of the motion-corrected fetal brain fMRI. We performed an extensive parameter study to investigate the effectiveness of motion estimation and present in this work benchmark metrics to quantify the effect of motion correction and regularised volumetric reconstruction approaches on functional connectivity computations. We demonstrate the proposed framework's ability to improve functional connectivity estimates, reproducibility and signal interpretability, which is clinically highly desirable for the establishment of prognostic noninvasive imaging biomarkers. The motion correction and volumetric reconstruction framework is made available as an open-source package of NiftyMIC.
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Artefactos , Imagen por Resonancia Magnética , Encéfalo/diagnóstico por imagen , Feto/diagnóstico por imagen , Humanos , Procesamiento de Imagen Asistido por Computador/métodos , Imagen por Resonancia Magnética/métodos , Movimiento (Física) , Reproducibilidad de los ResultadosRESUMEN
The removal of misfolded, ubiquitinated proteins is an essential part of the protein quality control. The ubiquitin-proteasome system (UPS) and autophagy are two interconnected pathways that mediate the degradation of such proteins. During autophagy, ubiquitinated proteins are clustered in a p62-dependent manner and are subsequently engulfed by autophagosomes. However, the nature of the protein substrates targeted for autophagy is unclear. Here, we developed a reconstituted system using purified components and show that p62 and ubiquitinated proteins spontaneously coalesce into larger clusters. Efficient cluster formation requires substrates modified with at least two ubiquitin chains longer than three moieties and is based on p62 filaments cross-linked by the substrates. The reaction is inhibited by free ubiquitin, K48-, and K63-linked ubiquitin chains, as well as by the autophagosomal marker LC3B, suggesting a tight cross talk with general proteostasis and autophagosome formation. Our study provides mechanistic insights on how substrates are channeled into autophagy.
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Autofagia/fisiología , Agregación Patológica de Proteínas/prevención & control , Proteínas de Unión al ARN/metabolismo , Proteínas Ubiquitinadas/metabolismo , Autofagosomas/fisiología , Línea Celular Tumoral , Humanos , Proteínas Asociadas a Microtúbulos/metabolismo , Agregación Patológica de Proteínas/patología , Pliegue de Proteína , Ubiquitina/metabolismoRESUMEN
OBJECTIVE: Clinical outcomes in multiple sclerosis (MS) are highly variable. We aim to determine the long-term clinical outcomes in MS, and to identify early prognostic features of these outcomes. METHODS: One hundred thirty-two people presenting with a clinically isolated syndrome were prospectively recruited between 1984 and 1987, and followed up clinically and radiologically 1, 5, 10, 14, 20, and now 30 years later. All available notes and magnetic resonance imaging scans were reviewed, and MS was defined according to the 2010 McDonald criteria. RESULTS: Clinical outcome data were obtained in 120 participants at 30 years. Eighty were known to have developed MS by 30 years. Expanded Disability Status Scale (EDSS) scores were available in 107 participants, of whom 77 had MS; 32 (42%) remained fully ambulatory (EDSS scores ≤3.5), all of whom had relapsing-remitting MS (RRMS), 3 (4%) had RRMS and EDSS scores >3.5, 26 (34%) had secondary progressive MS (all had EDSS scores >3.5), and MS contributed to death in 16 (20%). Of those with MS, 11 received disease-modifying therapy. The strongest early predictors (within 5 years of presentation) of secondary progressive MS at 30 years were presence of baseline infratentorial lesions and deep white matter lesions at 1 year. INTERPRETATION: Thirty years after onset, in a largely untreated cohort, there was a divergence of MS outcomes; some people accrued substantial disability early on, whereas others ran a more favorable long-term course. These outcomes could, in part, be predicted by radiological findings from within 1 year of first presentation. ANN NEUROL 2020;87:63-74.
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Enfermedades Desmielinizantes/epidemiología , Esclerosis Múltiple/epidemiología , Esclerosis Múltiple/patología , Adulto , Encéfalo/patología , Comorbilidad , Evaluación de la Discapacidad , Femenino , Estudios de Seguimiento , Humanos , Imagen por Resonancia Magnética , Masculino , Esclerosis Múltiple/diagnóstico , Neuroimagen , Valor Predictivo de las Pruebas , Pronóstico , Factores de Tiempo , Reino Unido/epidemiología , Adulto JovenRESUMEN
PURPOSE: A retrospective study was performed to study the effect of fetal surgery on brain development measured by MRI in fetuses with myelomeningocele (MMC). METHODS: MRI scans of 12 MMC fetuses before and after surgery were compared to 24 age-matched controls without central nervous system abnormalities. An automated super-resolution reconstruction technique generated isotropic brain volumes to mitigate 2D MRI fetal motion artefact. Unmyelinated white matter, cerebellum and ventricles were automatically segmented, and cerebral volume, shape and cortical folding were thereafter quantified. Biometric measures were calculated for cerebellar herniation level (CHL), clivus-supraocciput angle (CSO), transverse cerebellar diameter (TCD) and ventricular width (VW). Shape index (SI), a mathematical marker of gyrification, was derived. We compared cerebral volume, surface area and SI before and after MMC fetal surgery versus controls. We additionally identified any relationship between these outcomes and biometric measurements. RESULTS: MMC ventricular volume/week (mm3/week) increased after fetal surgery (median: 3699, interquartile range (IQR): 1651-5395) compared to controls (median: 648, IQR: 371-896); P = 0.015. The MMC SI is higher pre-operatively in all cerebral lobes in comparison to that in controls. Change in SI/week in MMC fetuses was higher in the left temporal lobe (median: 0.039, IQR: 0.021-0.054), left parietal lobe (median: 0.032, IQR: 0.023-0.039) and right occipital lobe (median: 0.027, IQR: 0.019-0.040) versus controls (P = 0.002 to 0.005). Ventricular volume (mm3) and VW (mm) (r = 0.64), cerebellar volume and TCD (r = 0.56) were moderately correlated. CONCLUSIONS: Following fetal myelomeningocele repair, brain volume, shape and SI were significantly different from normal in most cerebral layers. Morphological brain changes after fetal surgery are not limited to hindbrain herniation reversal. These findings may have neurocognitive outcome implications and require further evaluation.
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Meningomielocele , Disrafia Espinal , Encéfalo/diagnóstico por imagen , Encéfalo/cirugía , Feto , Humanos , Imagen por Resonancia Magnética , Meningomielocele/diagnóstico por imagen , Meningomielocele/cirugía , Estudios RetrospectivosRESUMEN
High-resolution volume reconstruction from multiple motion-corrupted stacks of 2D slices plays an increasing role for fetal brain Magnetic Resonance Imaging (MRI) studies. Currently existing reconstruction methods are time-consuming and often require user interactions to localize and extract the brain from several stacks of 2D slices. We propose a fully automatic framework for fetal brain reconstruction that consists of four stages: 1) fetal brain localization based on a coarse segmentation by a Convolutional Neural Network (CNN), 2) fine segmentation by another CNN trained with a multi-scale loss function, 3) novel, single-parameter outlier-robust super-resolution reconstruction, and 4) fast and automatic high-resolution visualization in standard anatomical space suitable for pathological brains. We validated our framework with images from fetuses with normal brains and with variable degrees of ventriculomegaly associated with open spina bifida, a congenital malformation affecting also the brain. Experiments show that each step of our proposed pipeline outperforms state-of-the-art methods in both segmentation and reconstruction comparisons including expert-reader quality assessments. The reconstruction results of our proposed method compare favorably with those obtained by manual, labor-intensive brain segmentation, which unlocks the potential use of automatic fetal brain reconstruction studies in clinical practice.
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Encéfalo/diagnóstico por imagen , Feto/diagnóstico por imagen , Hidrocefalia/diagnóstico por imagen , Procesamiento de Imagen Asistido por Computador/métodos , Imagen por Resonancia Magnética , Espina Bífida Quística/diagnóstico por imagen , Aprendizaje Profundo , Femenino , Terapias Fetales , Edad Gestacional , Humanos , Redes Neurales de la Computación , Embarazo , Espina Bífida Quística/cirugíaRESUMEN
PURPOSE: Magnetic resonance (MR) cholangiopancreatography (MRCP) is an established specialist method for imaging the upper abdomen and biliary/pancreatic ducts. Due to limitations of either MR image contrast or low through-plane resolution, patients may require further evaluation with contrast-enhanced computed tomography (CT) images. However, CT fails to offer the high tissue-ductal-vessel contrast-to-noise ratio available on T2-weighted MR imaging. METHODS: MR super-resolution reconstruction (SRR) frameworks have the potential to provide high-resolution visualizations from multiple low through-plane resolution single-shot T2-weighted (SST2W) images as currently used during MRCP studies. Here, we (i) optimize the source image acquisition protocols by establishing the ideal number and orientation of SST2W series for MRCP SRR generation, (ii) optimize post-processing protocols for two motion correction candidate frameworks for MRCP SRR, and (iii) perform an extensive validation of the overall potential of upper abdominal SRR, using four expert readers with subspeciality interest in hepato-pancreatico-biliary imaging. RESULTS: Obtained SRRs show demonstrable advantages over traditional SST2W MRCP data in terms of anatomical clarity and subjective radiologists' preference scores for a range of anatomical regions that are especially critical for the management of cancer patients. CONCLUSIONS: Our results underline the potential of using SRR alongside traditional MRCP data for improved clinical diagnosis.
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Abdomen/diagnóstico por imagen , Pancreatocolangiografía por Resonancia Magnética/métodos , Interpretación de Imagen Asistida por Computador/métodos , Adulto , Encéfalo/diagnóstico por imagen , Femenino , Voluntarios Sanos , Humanos , Aumento de la Imagen/métodos , Imagenología Tridimensional/métodos , Masculino , Proyectos PilotoRESUMEN
Lysosomes are the main degradative compartments of mammalian cells and serve as platforms for cellular nutrient signaling and sterol transport. The diverse functions of lysosomes and their adaptation to extracellular and intracellular cues are tightly linked to the spatiotemporally controlled synthesis, turnover and interconversion of lysosomal phosphoinositides, minor phospholipids that define membrane identity and couple membrane dynamics to cell signaling. How precisely lysosomal phosphoinositides act and which effector proteins within the lysosome membrane or at the lysosomal surface recognize them is only now beginning to emerge. Importantly, mutations in phosphoinositide metabolizing enzyme cause lysosomal dysfunction and are associated with numerous diseases ranging from neurodegeneration to cancer. Here, we discuss the phosphoinositides and phosphoinositide metabolizing enzymes implicated in lysosome function and homeostasis and outline perspectives for future research.
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Homeostasis , Lisosomas/metabolismo , Fosfatidilinositoles/metabolismo , Animales , Autofagosomas/metabolismo , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Transporte Biológico , Humanos , Membranas Intracelulares/metabolismo , Diana Mecanicista del Complejo 1 de la Rapamicina/metabolismo , Esteroles/metabolismoRESUMEN
Hospitals often hold historical MR image data printed on films without being able to make it accessible to modern image processing techniques. Having the possibility to recover geometrically consistent, volumetric images from scans acquired decades ago will enable more comprehensive, longitudinal studies to understand disease progressions. In this paper, we propose a consistent framework to reconstruct a volumetric representation from printed films holding thick single-slice brain MR image acquisitions dating back to the 1980's. We introduce a flexible framework based on semi-automatic slice extraction, followed by automated slice-to-volume registration with inter-slice transformation regularisation and slice intensity correction. Our algorithm is robust against numerous detrimental effects being present in archaic films. A subsequent, isotropic total variation deconvolution technique revitalises the visual appearance of the obtained volumes. We assess the accuracy and perform the validation of our reconstruction framework on a uniquely long-term MRI dataset where a ground-truth is available. This method will be used to facilitate a robust longitudinal analysis spanning 30 years of MRI scans.
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Algoritmos , Encéfalo/diagnóstico por imagen , Interpretación de Imagen Asistida por Computador/métodos , Imagenología Tridimensional/métodos , Neuroimagen/métodos , Conjuntos de Datos como Asunto , Humanos , Aumento de la Imagen/métodos , Estudios Longitudinales , Imagen por Resonancia Magnética , Película para Rayos XRESUMEN
Introduction: Hyperspectral imaging (HSI) has shown promise in the field of intra-operative imaging and tissue differentiation as it carries the capability to provide real-time information invisible to the naked eye whilst remaining label free. Previous iterations of intra-operative HSI systems have shown limitations, either due to carrying a large footprint limiting ease of use within the confines of a neurosurgical theater environment, having a slow image acquisition time, or by compromising spatial/spectral resolution in favor of improvements to the surgical workflow. Lightfield hyperspectral imaging is a novel technique that has the potential to facilitate video rate image acquisition whilst maintaining a high spectral resolution. Our pre-clinical and first-in-human studies (IDEAL 0 and 1, respectively) demonstrate the necessary steps leading to the first in-vivo use of a real-time lightfield hyperspectral system in neuro-oncology surgery. Methods: A lightfield hyperspectral camera (Cubert Ultris ×50) was integrated in a bespoke imaging system setup so that it could be safely adopted into the open neurosurgical workflow whilst maintaining sterility. Our system allowed the surgeon to capture in-vivo hyperspectral data (155 bands, 350-1,000 nm) at 1.5 Hz. Following successful implementation in a pre-clinical setup (IDEAL 0), our system was evaluated during brain tumor surgery in a single patient to remove a posterior fossa meningioma (IDEAL 1). Feedback from the theater team was analyzed and incorporated in a follow-up design aimed at implementing an IDEAL 2a study. Results: Focusing on our IDEAL 1 study results, hyperspectral information was acquired from the cerebellum and associated meningioma with minimal disruption to the neurosurgical workflow. To the best of our knowledge, this is the first demonstration of HSI acquisition with 100+ spectral bands at a frame rate over 1Hz in surgery. Discussion: This work demonstrated that a lightfield hyperspectral imaging system not only meets the design criteria and specifications outlined in an IDEAL-0 (pre-clinical) study, but also that it can translate into clinical practice as illustrated by a successful first in human study (IDEAL 1). This opens doors for further development and optimisation, given the increasing evidence that hyperspectral imaging can provide live, wide-field, and label-free intra-operative imaging and tissue differentiation.
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Purpose: Hyperspectral imaging shows promise for surgical applications to non-invasively provide spatially resolved, spectral information. For calibration purposes, a white reference image of a highly reflective Lambertian surface should be obtained under the same imaging conditions. Standard white references are not sterilizable and so are unsuitable for surgical environments. We demonstrate the necessity for in situ white references and address this by proposing a novel, sterile, synthetic reference construction algorithm. Approach: The use of references obtained at different distances and lighting conditions to the subject were examined. Spectral and color reconstructions were compared with standard measurements qualitatively and quantitatively, using ΔE and normalized RMSE, respectively. The algorithm forms a composite image from a video of a standard sterile ruler, whose imperfect reflectivity is compensated for. The reference is modeled as the product of independent spatial and spectral components, and a scalar factor accounting for gain, exposure, and light intensity. Evaluation of synthetic references against ideal but non-sterile references is performed using the same metrics alongside pixel-by-pixel errors. Finally, intraoperative integration is assessed though cadaveric experiments. Results: Improper white balancing leads to increases in all quantitative and qualitative errors. Synthetic references achieve median pixel-by-pixel errors lower than 6.5% and produce similar reconstructions and errors to an ideal reference. The algorithm integrated well into surgical workflow, achieving median pixel-by-pixel errors of 4.77% while maintaining good spectral and color reconstruction. Conclusions: We demonstrate the importance of in situ white referencing and present a novel synthetic referencing algorithm. This algorithm is suitable for surgery while maintaining the quality of classical data reconstruction.
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Images document scientific discoveries and are prevalent in modern biomedical research. Microscopy imaging in particular is currently undergoing rapid technological advancements. However for scientists wishing to publish the obtained images and image analyses results, there are to date no unified guidelines. Consequently, microscopy images and image data in publications may be unclear or difficult to interpret. Here we present community-developed checklists for preparing light microscopy images and image analysis for publications. These checklists offer authors, readers, and publishers key recommendations for image formatting and annotation, color selection, data availability, and for reporting image analysis workflows. The goal of our guidelines is to increase the clarity and reproducibility of image figures and thereby heighten the quality and explanatory power of microscopy data is in publications.
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Hyperspectral imaging is one of the most promising techniques for intraoperative tissue characterisation. Snapshot mosaic cameras, which can capture hyperspectral data in a single exposure, have the potential to make a real-time hyperspectral imaging system for surgical decision-making possible. However, optimal exploitation of the captured data requires solving an ill-posed demosaicking problem and applying additional spectral corrections. In this work, we propose a supervised learning-based image demosaicking algorithm for snapshot hyperspectral images. Due to the lack of publicly available medical images acquired with snapshot mosaic cameras, a synthetic image generation approach is proposed to simulate snapshot images from existing medical image datasets captured by high-resolution, but slow, hyperspectral imaging devices. Image reconstruction is achieved using convolutional neural networks for hyperspectral image super-resolution, followed by spectral correction using a sensor-specific calibration matrix. The results are evaluated both quantitatively and qualitatively, showing clear improvements in image quality compared to a baseline demosaicking method using linear interpolation. Moreover, the fast processing time of 45 ms of our algorithm to obtain super-resolved RGB or oxygenation saturation maps per image for a state-of-the-art snapshot mosaic camera demonstrates the potential for its seamless integration into real-time surgical hyperspectral imaging applications.
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Neuro-oncology surgery would benefit from detailed intraoperative tissue characterization provided by noncontact, contrast-agent-free, noninvasive optical imaging methods. In-depth knowledge of target tissue optical properties across a wide-wavelength spectrum could inform the design of optical imaging and computational methods to enable robust tissue analysis during surgery. We adapted a dual-beam integrating sphere to analyse small tissue samples and investigated ex vivo optical properties of five types of human brain tumour (meningioma, pituitary adenoma, schwannoma, low- and high-grade glioma) and nine different types of healthy brain tissue across a wavelength spectrum of 400 to 1800 nm. Fresh and frozen tissue samples were analysed. All tissue types demonstrated similar absorption spectra, but the reduced scattering coefficients of tumours show visible differences in the obtained optical spectrum compared to those of surrounding normal tissue. These results underline the potential of optical imaging technologies for intraoperative tissue characterization.