Morphology

形态学
  • 文章类型: Journal Article
    多年来,图像分析应用的经常被认为的局限性阻碍了此类系统作为第一线表征工具的广泛应用。图像分析,然而,在制药业中经历了显着的复苏,这得益于系统能力的发展以及科学家更充分地表征其颗粒的形态性质的愿望。颗粒形状以及尺寸的重要性现在被广泛认可。随着建模和模拟的使用越来越多,通过机器学习和人工智能的集成,在所获得数据的丰富性的推动下,图像分析的效用显着增加。这样的数据集提供了一种手段来规避依赖信息较少的描述符的要求,并能够朝着使用整体分布的方向发展。将改进的粒度和形状测量和描述与建模和模拟的进步相结合,可以改进方法来阐明颗粒和散装粉末性质之间的联系。除了改进描述输入材料的能力外,表征多组分系统中单一组分的方法为科学家提供了理解其材料在制造过程中如何变化的手段,从而提供了一种将最终剂型的行为与作用点处的颗粒性质联系起来的手段。目的是提供图像分析的概述,并通过小分子领域的其他方法的创新和能力更新读者。我们还将描述使用AI进行图像分析的改进分析。
    The often-perceived limitations of application of image analysis have for many years impeded the widespread application of such systems as first line characterisation tools. Image analysis has, however, undergone a notable resurgence in the pharmaceutical industry fuelled by developments system capabilities and the desire of scientists to characterize the morphological nature of their particles more adequately. The importance of particle shape as well as size is now widely acknowledged. With the increasing use of modelling and simulations, and ongoing developments though the integration of machine learning and artificial intelligence, the utility of image analysis is increasing significantly driven by the richness of the data obtained. Such datasets provide means to circumvent the requirement to rely on less informative descriptors and enable the move towards the use of whole distributions. Combining the improved particle size and shape measurement and description with advances in modelling and simulations is enabling improved means to elucidate the link between particle and bulk powder properties. In addition to improved capabilities to describe input materials, approaches to characterize single components within multicomponent systems are providing scientists means to understand how their material may change during manufacture thus providing a means to link the behaviour of final dosage forms with the particle properties at the point of action. The aim is to provide an overview of image analysis and update readers with innovations and capabilities to other methods in the small molecule arena. We will also describe the use of AI for the improved analysis using image analysis.
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  • 文章类型: Journal Article
    使用溶液燃烧程序来产生一系列Na3LaxSm1-x(PO4)2(x=0.01-0.15mol)纳米晶体,这些纳米晶体会产生温暖的深微红光。HR-TEM和X射线衍射检查均用于检查形态和结晶相分析。能量色散X射线分析(EDAX)批准元素检查。在近紫外线照射下,发光光谱在700nm波长处表现出良好的红橙色发射,与电子跃迁4G5/2→6H11/2对齐。根据Dexter的想法,在Sm3+离子组成达到6mol%之后发生的浓度猝灭是最近的相邻互连的原因。此外,辐射寿命的详细评估(0.7519ms),量子效率(77%),非辐射率(307.40),色温(3170K),色纯度(99.2%)和色坐标(0.652,0.338)。已经观察到的光学特性表明Sm3+掺杂的Na3La(PO4)2磷光体可以是改善WLED效率和颜色质量的良好选择。
    Solution combustion procedure was used to create a succession of Na3LaxSm1 - x(PO4)2 (x = 0.01-0.15 mol) nanocrystals that generate a warm deep reddish light. Both HR-TEM and X-ray diffraction examinations were used to examine the morphology and crystalline phase analysis. Energy-dispersive X-ray analysis (EDAX) approves the elemental examination. The luminescence spectrum exhibits a decent reddish-orange emission at 700 nm wavelength upon near-UV illumination, which aligns with the electronic transition 4G5/2 → 6H11/2. According to Dexter\'s idea, nearest neighbor interlinkages are responsible for the concentration quenching that occurs after the Sm3+ ion composition reaches 6 mol%. Additionally, a detailed evaluation of the radiative lifespan (0.7519 ms), quantum efficiency (77%), Non radiative rate (307.40), color temperature (3170 K), color purity (99.2%) and color coordinates (0.652, 0.338) was conducted. The optical characteristics that have been observed indicate that Sm3+ doped Na3La(PO4)2 phosphors could be a good option for improving WLED efficiency and color quality.
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  • 文章类型: Journal Article
    准确估计隐藻生物量,包括苔藓植物和地衣,对于理解它们的生态意义至关重要。该估计是基于密码的质量和体积之间的强相关性进行的。然而,由于它们的形态差异,质量-体积相关性在密码学之间有所不同。可以使用考虑基于形态相似性对密码进行分类的生命形式的模型来解决此问题。在这项研究中,我们调查了生命体模型是否提高了密码生物量估计的准确性。使用贝叶斯线性模型估计了每种生命体的密码质量-体积相关性。不同生命形式的线性模型的系数和截距不同,这归因于每种生命形式的形态特征。因此,生命形态模型可以通过结合形态差异来提高估计模型的准确性。然而,只考虑分类学差异的分类学模型(苔藓植物与地衣)表现出比生命形式模型更好的整体估计,可能是因为分类学模型能够捕获苔藓植物和地衣之间的系统差异。此外,这些模型可以减轻与形态变异相关的估计误差,而这些变异不能被生命形态类型充分表示.基于这些结果,我们建议适当使用估计模型。
    Accurate estimation of cryptogam biomass, encompassing bryophytes and lichens, is crucial for understanding their ecological significance. This estimation is conducted based on the strong correlations between mass and volume of cryptogams. However, mass-volume correlations vary among cryptogams because of their morphological differences. This problem can be solved using models that consider life forms that classify cryptogams based on morphological similarities. In this study, we investigated whether life form models improve cryptogam biomass estimation accuracy. The cryptogam mass-volume correlation of each life form was estimated using Bayesian linear models. The coefficients and intercepts of linear models differed between life forms, which was attributed to the morphological characteristics of each life form. Therefore, life form models can improve the accuracy of estimation models by incorporating morphological differences. However, taxonomic models that consider only the taxonomic difference (bryophytes vs lichens) demonstrated better overall estimation than the life form models, probably because of the ability of taxonomic models to capture systematic differences between bryophytes and lichens. Furthermore, these models may mitigate estimation errors related to morphological variations that cannot be adequately represented by life form types. Based on these results, we propose the appropriate use of estimation models.
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  • 文章类型: Journal Article
    长期饮酒如何导致持续性疼痛病理的机制尚不清楚。了解短期饮酒的早期事件如何降低非疼痛刺激的阈值,被描述为异常性疼痛可能被证明是谨慎的,以了解重要的启动机制。以前,我们观察到短期低剂量酒精摄入导致女性特异性异常性疼痛和脊髓背角小胶质细胞活化增加.其他文献描述了慢性乙醇暴露如何激活Toll样受体4(TLR4)以引发炎症反应。TLR4在许多细胞类型上表达,我们的目的是研究在短期/低剂量酒精模式下,小胶质细胞上的TLR4是否足以增强异常性疼痛。我们的研究使用了一种新的遗传模型,其中TLR4的表达通过引入一个浮动的转录阻断剂(TLR4-null背景(TLR4LoxTB))从整个身体中去除,然后通过用Cx3CR1:CreERT2动物饲养TLR4LoxTB动物来限制小胶质细胞。正如以前报道的那样,单独服用乙醇14天后,我们没有观察到疼痛行为增加。然而,我们在野生型和小胶质细胞-TLR4限制性雌性小鼠足底注射亚阈值剂量前列腺素E2(PGE2)后3小时观察到显著的启动效应.我们还观察到女性特异性向促炎表型的显着转变以及腰背角小胶质细胞的形态变化。对与疼痛引发相关的神经元亚型的研究表明,雌性小鼠背角PKCγ中间神经元中c-Fos和FosB活性的增加直接对应于小胶质细胞活性的增加。这项研究揭示了TLR4在非病理性饮酒者疼痛诱导性二态中的细胞和女性特异性作用。
    The mechanisms of how long-term alcohol use can lead to persistent pain pathology are unclear. Understanding how earlier events of short-term alcohol use can lower the threshold of non-painful stimuli, described as allodynia could prove prudent to understand important initiating mechanisms. Previously, we observed that short-term low-dose alcohol intake induced female-specific allodynia and increased microglial activation in the spinal cord dorsal horn. Other literature describes how chronic ethanol exposure activates Toll-like receptor 4 (TLR4) to initiate inflammatory responses. TLR4 is expressed on many cell types, and we aimed to investigate whether TLR4 on microglia is sufficient to potentiate allodynia during a short-term/low-dose alcohol paradigm. Our study used a novel genetic model where TLR4 expression is removed from the entire body by introducing a floxed transcriptional blocker (TLR4-null background (TLR4LoxTB)), then restricted to microglia by breeding TLR4LoxTB animals with Cx3CR1:CreERT2 animals. As previously reported, after 14 days of ethanol administration alone, we observed no increased pain behavior. However, we observed significant priming effects 3 h post intraplantar injection of a subthreshold dose of prostaglandin E2 (PGE2) in wild-type and microglia-TLR4 restricted female mice. We also observed a significant female-specific shift to pro-inflammatory phenotype and morphological changes in microglia of the lumbar dorsal horn. Investigations in pain priming-associated neuronal subtypes showed an increase of c-Fos and FosB activity in PKCγ interneurons in the dorsal horn of female mice directly corresponding to increased microglial activity. This study uncovers cell- and female-specific roles of TLR4 in sexual dimorphisms in pain induction among non-pathological drinkers.
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  • 文章类型: Journal Article
    细菌的生长和分裂依赖于形态发生复合物的复杂调节来重塑细胞包膜而不损害包膜完整性。近年来,在了解细胞壁代谢酶的调节方面取得了重大进展。然而,其他细胞包膜成分也在形态发生中起作用。在低渗透压环境中保护包膜完整性所需的主要因素是OpgH,渗透调节周质葡聚糖(OPGs)的合酶。这里,我们证明OpgH在α-蛋白细菌中必不可少。出乎意料的是,耗尽OpgH或尝试用催化死亡的OpgH变体进行补充导致显著的不对称膨胀和细胞裂解。这些形状缺陷伴随着细胞壁合成的减少和形态发生复合物的错误定位。有趣的是,CenKR双组分系统的过度激活与α-蛋白细菌表型中的细胞包膜应激稳态有关,与OpgH耗竭相关的形态发生缺陷。这些扰动中的每一个都导致延长体蛋白水平的增加,MreB,以及分裂体蛋白FtsZ和MipZ以及OpgH的水平降低,本身。在CenKR过度激活期间,OpgH的组成型产生可防止细胞膨胀,但是细胞仍然表现出形态发生缺陷。我们建议OPG消耗激活CenKR,导致细胞包膜相关基因表达的变化,但是OPGs也对形态发生产生不依赖CenKR的影响。我们的数据确立了OpgH同系物在形态发生中的令人惊讶的功能,并揭示了OpgH在维持杆菌属细胞形态中的重要作用。重要细菌必须以严格调节的方式合成和强化细胞包膜,以协调生长和适应。渗透调节的周质葡聚糖(OPGs)是重要的,但却知之甚少,革兰氏阴性细胞包膜的成分,有助于包膜完整性并防止渗透应激。这里,我们确定OPG合酶OpgH发挥了令人惊讶的作用,在结球菌形态发生中的重要作用。OpgH的丢失通过对形态发生复合物的定位和活性的错误调节而导致不对称的细胞膨胀和裂解。CenKR双组分系统的过度激活参与包膜稳态表型的OpgH耗竭,表明OpgH的消耗激活了CenKR。因为细胞包膜的完整性对细菌的生存至关重要,了解OpgH活性如何有助于形态发生和包膜完整性的维持可能有助于抗生素疗法的开发。
    Bacterial growth and division rely on intricate regulation of morphogenetic complexes to remodel the cell envelope without compromising envelope integrity. Significant progress has been made in recent years towards understanding the regulation of cell wall metabolic enzymes. However, other cell envelope components play a role in morphogenesis as well. A primary factor required to protect envelope integrity in low osmolarity environments is OpgH, the synthase of osmoregulated periplasmic glucans (OPGs). Here, we demonstrate that OpgH is essential in the α-proteobacterium Caulobacter crescentus. Unexpectedly, depletion of OpgH or attempted complementation with a catalytically dead OpgH variant results in striking asymmetric bulging and cell lysis. These shape defects are accompanied by reduced cell wall synthesis and mislocalization of morphogenetic complexes. Interestingly, overactivation of the CenKR two-component system that has been implicated in cell envelope stress homeostasis in α-proteobacteria phenocopies the morphogenetic defects associated with OpgH depletion. Each of these perturbations leads to an increase in the levels of the elongasome protein, MreB, and decreases in the levels of divisome proteins FtsZ and MipZ as well as OpgH, itself. Constitutive production of OpgH during CenKR overactivation prevents cell bulging, but cells still exhibit morphogenetic defects. We propose that OPG depletion activates CenKR, leading to changes in the expression of cell envelope-related genes, but that OPGs also exert CenKR-independent effects on morphogenesis. Our data establish a surprising function for an OpgH homolog in morphogenesis and reveal an essential role of OpgH in maintaining cell morphology in Caulobacter.IMPORTANCEBacteria must synthesize and fortify the cell envelope in a tightly regulated manner to orchestrate growth and adaptation. Osmoregulated periplasmic glucans (OPGs) are important, but poorly understood, constituents of Gram-negative cell envelopes that contribute to envelope integrity and protect against osmotic stress. Here, we determined that the OPG synthase OpgH plays a surprising, essential role in morphogenesis in Caulobacter crescentus. Loss of OpgH causes asymmetric cell bulging and lysis via misregulation of the localization and activity of morphogenetic complexes. Overactivation of the CenKR two-component system involved in envelope homeostasis phenocopies OpgH depletion, suggesting that depletion of OpgH activates CenKR. Because cell envelope integrity is critical for bacterial survival, understanding how OpgH activity contributes to morphogenesis and maintenance of envelope integrity could aid in the development of antibiotic therapies.
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  • 文章类型: Journal Article
    双酚S(BPS)广泛用于制造产品,增加心血管疾病的风险。肥胖和BPS之间的关联对心脏预后的影响仍然未知。将雄性C57BL/6小鼠分为标准饮食(SC;15kJ/g),标准饮食+BPS(SCB),高脂饮食(HF;21kJ/g),和高脂饮食+BPS(HFB)。超过12周,两组通过饮用水(剂量:25μg/Kg/天)和/或HF饮食暴露于BPS。我们评估:体重(BM),总胆固醇,收缩压(SBP),左心室(LV)质量,和心脏重塑。在SCB组中,BM,总胆固醇,与SC组相关,SBP增加。在HF和HFB组中,这些参数高于SC和SCB组.左心室质量和壁厚增加证明了心肌肥厚。与SC组相比,所有组的ANP蛋白表达。只有HFB组比SCB和HF组有更厚的LV壁,与SC和SCB组相比,心肌细胞面积增加。关于心脏纤维化,SCB,HF,和HFB组呈现较高的间质胶原面积,TGFβ,而α-SMA蛋白表达高于SC组。仅HF和HFB组的血管周围胶原面积比SC组增加。较高的IL-6,TNFα,和CD11c卵白表达在一切组均比SC组证明的炎症。与SC组相比,所有组的CD36和PPARα蛋白表达均升高,但只有HF和HFB组比SC组促进心脏脂肪变性,perilipin5蛋白表达增加。单独BPS暴露促进病理性同心性肥大的心脏重塑,纤维化,和炎症。当与BPS相关时,饮食诱导的重塑会加剧,有明显的肥大,除了纤维化,炎症,和脂质积累。
    Bisphenol S (BPS) is widely used in the manufacture products and increase the risk of cardiovascular diseases. The effect of the association between obesity and BPS on cardiac outcomes is still unknown. Male C57BL/6 mice were divided into standard chow diet (SC; 15 kJ/g), standard chow diet + BPS (SCB), high-fat diet (HF; 21 kJ/g), and high-fat diet + BPS (HFB). Over 12 weeks, the groups were exposed to BPS through drinking water (dose: 25 μg/Kg/day) and/or a HF diet. We evaluated: body mass (BM), total cholesterol, systolic blood pressure (SBP), left ventricle (LV) mass, and cardiac remodeling. In the SCB group, BM, total cholesterol, and SBP increase were augmented in relation to the SC group. In the HF and HFB groups, these parameters were higher than in the SC and SCB groups. Cardiac hypertrophy was evidenced by augmented LV mass and wall thickness, and ANP protein expression in all groups in comparison to the SC group. Only the HFB group had a thicker LV wall than SCB and HF groups, and increased cardiomyocyte area when compared with SC and SCB groups. Concerning cardiac fibrosis, SCB, HF, and HFB groups presented higher interstitial collagen area, TGFβ, and α-SMA protein expression than the SC group. Perivascular collagen area was increased only in the HF and HFB groups than SC group. Higher IL-6, TNFα, and CD11c protein expression in all groups than the SC group evidenced inflammation. All groups had elevated CD36 and PPARα protein expression in relation to the SC group, but only HF and HFB groups promoted cardiac steatosis with increased perilipin 5 protein expression than the SC group. BPS exposure alone promoted cardiac remodeling with pathological concentric hypertrophy, fibrosis, and inflammation. Diet-induced remodeling is aggravated when associated with BPS, with marked hypertrophy, alongside fibrosis, inflammation, and lipid accumulation.
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  • 文章类型: Journal Article
    本研究探索了一维系统中光化学析氢催化剂的最佳形态。在精确定义的CdSe@CdS点中棒上进行金属尖端的系统工程,以控制形态,composition,和两个因素。结果产生了一个优化的配置,具有粗糙Pt表面的Au-Pt核-壳结构(Au@r-Pt),量子效率显着提高了五倍,在455nm下达到86%,在可见光和AM1.5G辐射条件下具有优异的氢析出率,并具有延长的稳定性。使用光电化学和时间分辨测量的动力学研究表明,尖端上的电荷分离状态的更大程度和更长的寿命,以及高能表面上的快速水还原动力学。这种方法揭示了助催化剂在混合光催化体系中实现高性能的关键作用。
    This study explores the optimal morphology of photochemical hydrogen evolution catalysts in a one-dimensional system. Systematic engineering of metal tips on precisely defined CdSe@CdS dot-in-rods is conducted to exert control over morphology, composition, and both factors. The outcome yields an optimized configuration, a Au-Pt core-shell structure with a rough Pt surface (Au@r-Pt), which exhibits a remarkable fivefold increase in quantum efficiency, reaching 86% at 455 nm and superior hydrogen evolution rates under visible and AM1.5G irradiation conditions with prolonged stability. Kinetic investigations using photoelectrochemical and time-resolved measurements demonstrate a greater extent and extended lifetime of the charge-separated state on the tips as well as rapid water reduction kinetics on high-energy surfaces. This approach sheds light on the critical role of cocatalysts in hybrid photocatalytic systems for achieving high performance.
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  • 文章类型: Journal Article
    非血栓性髂静脉病变(NIVL)是左下肢慢性静脉功能不全(CVI)和左下肢静脉曲张治疗后症状复发的重要原因。本研究的目的是探讨NIVL患者髂静脉的血流动力学和形态学特征。狭窄的左髂总静脉(LCIV)段的尾端压力,局部血流速度,狭窄段的时间平均壁切应力与临床CVI分类呈正相关(R=0.92,p<0.001;R=0.94,p<0.001;R=0.87,p<0.001),相对保留时间呈负相关(R=-0.94,p<0.001)。狭窄段两端的压差(ΔP)和狭窄段与尾端的速度差(ΔV)与临床分型呈正相关(R=0.92,p<0.001;R=0.9,p<0.001)。狭窄LCIV节段的横截面积狭窄率和长度与临床分型呈正相关(R=0.93,p<0.001;R=0.63,p<0.001)。结果表明,对LCIV狭窄段的血流动力学评估可以有效地描述血流紊乱,可能反映髂静脉狭窄的程度。血流动力学指标与临床CVI症状的严重程度相关。
    Nonthrombotic iliac vein lesions (NIVLs) are significant causes of chronic venous insufficiency (CVI) in the left lower limb and symptom recurrence following left lower limb varicose vein treatment. The goal of this study was to explore the haemodynamic and morphological characteristics of iliac veins in patients with NIVLs. Pressure at the caudal end of the stenotic left common iliac vein (LCIV) segment, local blood flow velocity, and time-averaged wall shear stress in the stenotic segment exhibited positive correlations with the clinical CVI classification (R = 0.92, p < 0.001; R = 0.94, p < 0.001; R = 0.87, p < 0.001), while the relative retention time showed a negative correlation (R = -0.94, p < 0.001). The pressure difference (∆P) between the two ends of the stenotic segment and the velocity difference (∆V) between the stenotic segment and the caudal end were positively correlated with the clinical classification (R = 0.92, p < 0.001; R = 0.9, p < 0.001). The cross-sectional area stenosis rate and length of the stenotic LCIV segment were positively correlated with the clinical classification (R = 0.93, p < 0.001; R = 0.63, p < 0.001). The results suggest that haemodynamic assessment of the iliac vein could effectively portray blood flow disturbances in stenotic segments of the LCIV, potentially reflecting the degree of iliac vein stenosis. Haemodynamic indicators are correlated with the severity of clinical CVI symptoms.
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  • 文章类型: Journal Article
    Several species of the worldwide distributed genus Calliphora Robineau-Desvoidy (Insecta, Diptera, Calliphoridae) are medically important vectors and agents of myiasis. Furthermore, these flies are relevant in forensics because they are found in corpses. Information regarding the taxonomy, bionomics and distribution of Calliphora species endemic to South America, including Calliphora lopesi Mello, is scarce. To expand knowledge on C. lopesi, this study presents descriptions of eggs, larvae, puparia and developmental data at 14, 17, 20, 23 and 26 ± 1°C for the first time. Adult flies were collected from the field and kept in the laboratory to obtain samples for morphological and biological studies. Immatures were examined using light and scanning electron microscopy. To assess the growth rate, 10 specimens from each temperature group were randomly removed from the diet and weighed every 24 h from larval hatching until pupation. The minimum developmental threshold, thermal constant and linear development-rate equations were calculated for each stage. Considering weight gain records and survival rates, the optimum temperature for the development of C. lopesi ranges from 23 to 26°C. A key to third-instar larvae of known Neotropical species of Calliphora was also provided to assist in identification. The information provided in this study should be useful in expanding knowledge about Neotropical Calliphoridae species of forensic importance.
    Várias espécies do gênero Calliphora Robineau‐Desvoidy (Insecta, Diptera, Calliphoridae), distribuídas mundialmente, são vetores e agentes causadoras de miíases clinicamente importantes. Além disso, tais moscas são relevantes no âmbito forense porque são encontradas em cadáveres. Informações sobre a taxonomia, bionomia e distribuição de espécies de Calliphora endêmicas da América do Sul, incluindo Calliphora lopesi Mello, ainda são escassas. Para ampliar o conhecimento sobre a espécie C. lopesi, este estudo apresenta pela primeira vez as descrições de ovos, larvas e pupários, assim como dados sobre o desenvolvimento de imaturos a 14, 17, 20, 23 e 26 ± 1°C. Moscas adultas foram coletadas no campo e mantidas em laboratório para obtenção de amostras para estudos morfológicos e biológicos. Os imaturos foram examinados utilizando microscopia óptica e eletrônica de varredura. Para avaliar a taxa de crescimento, 10 espécimes de cada grupo de temperatura foram retirados aleatoriamente do substrato em que se alimentavam e pesados a cada 24 horas, desde a eclosão das larvas até a pupariação. O limiar mínimo de desenvolvimento, a constante térmica e as equações lineares da taxa de desenvolvimento foram calculados para cada estágio. Levando em consideração os registros de ganho de peso e taxas de sobrevivência, a temperatura ótima para o desenvolvimento de C. lopesi varia de 23 a 26°C. Uma chave para larvas de terceiro estádio de espécies neotropicais conhecidas de Calliphora também está sendo disponibilizada para auxiliar na identificação. Espera‐se que as informações fornecidas neste estudo possam ser úteis para ampliar o conhecimento sobre espécies Neotropicais de Calliphoridae de importância forense.
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  • 文章类型: Journal Article
    神经元包含三个隔室,索马,长轴突,和树突,具有不同的能量和生化要求。线粒体的特征在所有区室和调节神经元的活动和生存,包括能量产生和钙缓冲以及其他作用,包括促凋亡信号和类固醇合成。它们的动态性使它们能够响应不断变化的能量和生化需求而经历不断的融合和裂变事件。这些事件,称为线粒体动力学,影响它们的形态和各种三维(3D)形态存在于神经元线粒体网络中。在衰老和常见的神经退行性疾病中,形态学特征的扭曲以及线粒体功能障碍可能始于神经元体细胞。然而,3D形态学不能在平面中全面检查,二维(2D)图像。这突出了需要在体积数据内分割线粒体,以提供支持健康和患病神经元内线粒体动力学和线粒体自噬的过程的代表性快照。自动高分辨率体积成像方法的出现,如串行块扫描电子显微镜(SBF-SEM)以及图像软件包的范围允许执行这一点。我们描述并评估了一种方法,该方法用于从SBF-SEM图像堆栈中随机生成的神经元体细胞感兴趣区域中随机采样线粒体并手动分割其整个形态。然后可以使用这些3D重建来生成关于线粒体和细胞形态的定量数据。我们进一步描述了宏的使用,该宏自动解剖体细胞并将3D线粒体定位到创建的子区域中。
    Neurons contain three compartments, the soma, long axon, and dendrites, which have distinct energetic and biochemical requirements. Mitochondria feature in all compartments and regulate neuronal activity and survival, including energy generation and calcium buffering alongside other roles including proapoptotic signaling and steroid synthesis. Their dynamicity allows them to undergo constant fusion and fission events in response to the changing energy and biochemical requirements. These events, termed mitochondrial dynamics, impact their morphology and a variety of three-dimensional (3D) morphologies exist within the neuronal mitochondrial network. Distortions in the morphological profile alongside mitochondrial dysfunction may begin in the neuronal soma in ageing and common neurodegenerative disorders. However, 3D morphology cannot be comprehensively examined in flat, two-dimensional (2D) images. This highlights a need to segment mitochondria within volume data to provide a representative snapshot of the processes underpinning mitochondrial dynamics and mitophagy within healthy and diseased neurons. The advent of automated high-resolution volumetric imaging methods such as Serial Block Face Scanning Electron Microscopy (SBF-SEM) as well as the range of image software packages allow this to be performed.We describe and evaluate a method for randomly sampling mitochondria and manually segmenting their whole morphologies within randomly generated regions of interest of the neuronal soma from SBF-SEM image stacks. These 3D reconstructions can then be used to generate quantitative data about mitochondrial and cellular morphologies. We further describe the use of a macro that automatically dissects the soma and localizes 3D mitochondria into the subregions created.
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