Contemporaneous 3D characterization of acute and chronic myocardial I/R injury and response

Merz, Simon F.; Korste, Sebastian; Bornemann, Lea; Michel, Lars; Stock, Pia; Squire, Anthony; Soun, Camille; Engel, Daniel R.; Detzer, Julia; Lörchner, Holger; Hermann, Dirk M.; Kamler, Markus; Klode, Joachim; Hendgen‐Cotta, Ulrike B.; Rassaf, Tienush; Gunzer, Matthias; Totzeck, Matthias

doi:10.1038/s41467-019-10338-2

Cited by 67 publications

(85 citation statements)

References 48 publications

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“…By combining imaging modalities, our method could visualise and analyse spatiotemporal differences in 3D, and would allow identification of injury-induced changes remote from injury site, especially for large animal tissues [47]. These approaches have already been attempted in renal and cardiac injuries in adult mice, demonstrating that 3D approaches can inform regional differences in tissue repair [48,49]. Hence, our proposed protocol could be scaled down for smaller samples, such as mouse Achilles tendon, to investigate regional differences in cell-ECM interactions in murine models of tendon injury using 3D whole-mount preparation.…”

Section: Discussionmentioning

confidence: 99%

Bimodal Whole-Mount Imaging of Tendon Using Confocal Microscopy and X-ray Micro-Computed Tomography

et al. 2020

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Background: Three-dimensional imaging modalities for optically dense connective tissues such as tendons are limited and typically have a single imaging methodological endpoint. Here, we have developed a bimodal procedure utilising fluorescence-based confocal microscopy and x-ray micro-computed tomography for the imaging of adult tendons to visualise and analyse extracellular sub-structure and cellular composition in small and large animal species. Results: Using fluorescent immunolabelling and optical clearing, we visualised the expression of the novel crossspecies marker of tendon basement membrane, laminin-α4 in 3D throughout whole rat Achilles tendons and equine superficial digital flexor tendon 5 mm segments. This revealed a complex network of laminin-α4 within the tendon core that predominantly localises to the interfascicular matrix compartment. Furthermore, we implemented a chemical drying process capable of creating contrast densities enabling visualisation and quantification of both fascicular and interfascicular matrix volume and thickness by x-ray micro-computed tomography. We also demonstrated that both modalities can be combined using reverse clarification of fluorescently labelled tissues prior to chemical drying to enable bimodal imaging of a single sample. Conclusions: Whole-mount imaging of tendon allowed us to identify the presence of an extensive network of laminin-α4 within tendon, the complexity of which cannot be appreciated using traditional 2D imaging techniques. Creating contrast for x-ray micro-computed tomography imaging of tendon using chemical drying is not only simple and rapid, but also markedly improves on previously published methods. Combining these methods provides the ability to gain spatio-temporal information and quantify tendon substructures to elucidate the relationship between morphology and function.

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Section: Discussionmentioning

confidence: 99%

Bimodal Whole-Mount Imaging of Tendon Using Confocal Microscopy and X-ray Micro-Computed Tomography

et al. 2020

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“…Among the various techniques available, immunolabeling-enabled three-dimensional imaging of solvent-cleared organs (iDISCO) has emerged as a simple and rapid method for achieving good antibody penetration and tissue clearing (Liebmann et al, 2016; Renier et al, 2014; Renier et al, 2016). iDISCO–LSFM is particularly useful for CNS research as the low brain anatomical variation between individual mice allows scanned brains to be mapped onto a common coordinated framework (CCF) for subsequent unbiased quantitative image analysis of immune-labeled molecular targets (Detrez et al, 2019; Hasegawa et al, 2019; Henning et al, 2019; Kjaergaard et al, 2019; Liebmann et al, 2016; Merz et al, 2019; Mzinza et al, 2018; Renier et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Quantitative whole-brain 3D imaging of tyrosine hydroxylase-labeled neuron architecture in the mouse MPTP model of Parkinson's disease

Roostalu

Salinas

Thorbek

et al. 2019

Disease Models &Amp; Mechanisms

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Parkinson's disease (PD) is a basal ganglia movement disorder characterized by progressive degeneration of the nigrostriatal dopaminergic system. Immunohistochemical methods have been widely used for characterization of dopaminergic neuronal injury in animal models of PD, including the MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) mouse model. However, conventional immunohistochemical techniques applied to tissue sections have inherent limitations with respect to loss of 3D resolution, yielding insufficient information on the architecture of the dopaminergic system. To provide a more comprehensive and non-biased map of MPTP-induced changes in central dopaminergic pathways, we used iDISCO immunolabeling, light-sheet fluorescence microscopy (LSFM) and deep-learning computational methods for whole-brain three-dimensional visualization and automated quantitation of tyrosine hydroxylase (TH)-positive neurons in the adult mouse brain. Mice terminated 7 days after acute MPTP administration demonstrated widespread alterations in TH expression. Compared to vehicle controls, MPTP-dosed mice showed a significant loss of TH-positive neurons in the substantia nigra pars compacta and ventral tegmental area. Also, MPTP dosing reduced overall TH signal intensity in basal ganglia nuclei, i.e. the substantia nigra, caudate-putamen, globus pallidus and subthalamic nucleus. In contrast, increased TH signal intensity was predominantly observed in limbic regions, including several subdivisions of the amygdala and hypothalamus. In conclusion, mouse whole-brain 3D imaging is ideal for unbiased automated counting and densitometric analysis of TH-positive cells. The LSFM–deep learning pipeline tracked brain-wide changes in catecholaminergic pathways in the MPTP mouse model of PD, and may be applied for preclinical characterization of compounds targeting dopaminergic neurotransmission.

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“…Importantly in the cardiovascular field, Merz et al . successfully imaged a sample of human atrium at single-cell resolution 38 . One consideration during imaging of human tissue is the sample size that can be successfully cleared and fit into the sample chamber.…”

Section: Discussionmentioning

confidence: 99%

Light Sheet Fluorescence Microscopy as a New Method for Unbiased Three-Dimensional Analysis of Vascular Injury

Buglak

Lucitti

Ariel

et al. 2020

Preprint

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25Aims. Assessment of preclinical models of vascular disease are paramount in the successful 26 translation of novel treatments. The results of these models have traditionally relied on 2-D 27 histological methodologies. Light sheet fluorescence microscopy (LSFM) is an imaging platform 28 that allows for 3-D visualization of whole organs and tissues. In this study, we describe an 29 improved methodological approach utilizing LSFM for imaging of preclinical vascular injury 30 models while minimizing analysis bias. 31Methods and Results. The rat carotid artery segmental pressure-controlled balloon injury and 32 mouse carotid artery ligation injury were performed. Arteries were harvested and processed for 33 LSFM imaging and 3-D analysis, as well as for 2-D area histological analysis. Artery processing 34 for LSFM imaging did not induce vessel shrinkage or expansion, and was reversible by 35 rehydrating the artery, allowing for subsequent sectioning and histological staining a posteriori. 36By generating a volumetric visualization along the length of the arteries, LSFM imaging provided 37 different analysis modalities including volumetric, area, and radial parameters. Thus, LSFM-38 imaged arteries provided more precise measurements compared to classic histological analysis. 39Furthermore, LSFM provided additional information as compared to 2-D analysis in demonstrating 40 remodeling of the arterial media in regions of hyperplasia and periadventitial neovascularization 41 around the ligated mouse artery. 42 Conclusions. LSFM provides a novel and robust 3-D imaging platform for visualizing and 43quantifying arterial injury in preclinical models. When compared with classic histology, LSFM 44 outperformed traditional methods in precision and quantitative capabilities. LSFM allows for more 45 comprehensive quantitation as compared to traditional histological methodologies, while 46 minimizing user bias associated with area analysis of alternating, 2-D histological artery cross-47 Translational Perspective 49A more reproducible and robust quantitation of vascular pathology in preclinical models is 50 necessary to accelerate translational discovery. Current methodology to assess vascular disease 51 has significant limitations. The methodology described herein employs a modern imaging 52 modality, light sheet fluorescence microscopy (LSFM), to improve assessment of established 53 preclinical vascular injury models. LSFM provides more comprehensive and precise analysis 54 capabilities than classical histological approaches. Hence, LSFM applied to vascular research 55 has the potential to drive new basic discoveries, and ultimately translation of novel therapies. 56 4 I. Introduction 57Cardiovascular disease (CVD) is the leading cause of death and disability in the world 1, 2 . 58Atherosclerosis is the major underlying cause of most CVD. Severe, symptomatic atherosclerosis 59 is treated by percutaneous or surgical revascularization, and the long-term success of both 60 approaches is limited by arterial restenosis 3 . Despite ...

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Contemporaneous 3D characterization of acute and chronic myocardial I/R injury and response

Cited by 67 publications

References 48 publications

Bimodal Whole-Mount Imaging of Tendon Using Confocal Microscopy and X-ray Micro-Computed Tomography

Bimodal Whole-Mount Imaging of Tendon Using Confocal Microscopy and X-ray Micro-Computed Tomography

Quantitative whole-brain 3D imaging of tyrosine hydroxylase-labeled neuron architecture in the mouse MPTP model of Parkinson's disease

Light Sheet Fluorescence Microscopy as a New Method for Unbiased Three-Dimensional Analysis of Vascular Injury

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