Angéline Geiser, Susan Currie, Hadi Al-Hasani, Alexandra Chadt, Gail McConnell, Gwyn W Gould
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引用次数: 0
摘要
细胞异质性是公认的组织特征,包括光学成像在内的多种方法都揭示了细胞转录和代谢的多样性。然而,高分辨率成像所需的高倍物镜只能提供小层组织的信息,这可能导致细胞统计不准确。因此,人们对一种成像方式的需求尚未得到满足,这种成像方式可以提供三维完整组织样本中详细的分子和细胞信息。我们以 GFP 标记的 GLUT4 作为概念验证,在此介绍一种新颖的光学中间镜方法,它能在完整小鼠心脏的超厚切片(5 mm x 5 mm x 3 mm)中精确测量 GLUT4 在心肌特定解剖结构中的空间位置。我们揭示了不同的 GLUT4 在心肌壁上的分布模式,并强调了 GLUT4 表达水平在高脂饮食喂养下的特定变化,我们还发现了表达模式的性别差异。这种方法适用于任何可在光镜下标记的目标,也适用于需要同时考虑器官结构和细胞细节的其他复杂组织。
A novel 3D imaging approach for quantification of GLUT4 levels across the intact myocardium.
Cellular heterogeneity is a well-accepted feature of tissues, and both transcriptional and metabolic diversity have been revealed by numerous approaches, including optical imaging. However, the high magnification objective lenses needed for high-resolution imaging provides information from only small layers of tissue, which can result in poor cell statistics. There is therefore an unmet need for an imaging modality that can provide detailed molecular and cellular insight within intact tissue samples in 3D. Using GFP-tagged GLUT4 as proof of concept, we present here a novel optical mesoscopy approach that allows precise measurement of the spatial location of GLUT4 within specific anatomical structures across the myocardium in ultrathick sections (5 mm×5 mm×3 mm) of intact mouse heart. We reveal distinct GLUT4 distribution patterns across cardiac walls and highlight specific changes in GLUT4 expression levels in response to high fat diet-feeding, and we identify sex-dependent differences in expression patterns. This method is applicable to any target that can be labelled for light microscopy, and to other complex tissues when organ structure needs to be considered simultaneously with cellular detail.