Perfusion staining methods for visualization of the intact microvascular networks in whole mount skeletal muscle preparations.

IF 1.8 4区医学 Q3 PERIPHERAL VASCULAR DISEASE Journal of Vascular Research Pub Date : 2024-11-19 DOI:10.1159/000542663

Barbara M Hyde-Lay, Mackenzie E Charter, Coral L Murrant

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Abstract

Introduction: Visualization of the intact microvascular network in skeletal muscle requires labeling the entire network in whole mount preparations where muscle fibre length can be set to near optimal but the tools to do this are not clear.

Methods: We intravascularly injected CD-1 mice with different fluorescently labelled lectins (fluorescent isolectin GS-IB4 (ISO), wheat germ agglutinin (WGA), lycopersicon esculentum (tomato) (LYCO) or FITC-labelled gel. Soleus, extensor digitorum longus, diaphragm, gluteus maximus and cremaster muscles were excised, pinned at optimal sarcomere length and viewed using fluorescence microscopy.

Results: WGA and LYCO were the effective at labeling the entire vascular network with WGA labeling capillaries more bright. ISO labelled the arteriolar vasculature and early segments of the capillaries but not the full length of the capillaries or the venular network. FITC-labelled gel was effective at labelling the microvascular network but not all small vessels were consistently labelled. The pattern of staining for each labelling method was similar across all muscle fibre-types tested.

Conclusions: WGA was optimal for perfusion labeling and visualization of the intact microvascular network in whole mount skeletal muscle preparations and can be used in combination with ISO to distinguish the arteriolar and venous sides of the network.

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灌注染色法，用于观察整块骨骼肌制备物中完整的微血管网络。

简介：骨骼肌中完整微血管网络的可视化需要在整个装片制备过程中标记整个网络，在此过程中肌肉纤维长度可设置为接近最佳值，但实现这一目标的工具尚不明确：骨骼肌中完整微血管网络的可视化需要在整体装片制备中标记整个网络，在整体装片制备中，肌肉纤维长度可设置为接近最佳值，但实现这一目标的工具尚不明确：我们向 CD-1 小鼠血管内注射了不同的荧光标记凝集素（荧光异选择素 GS-IB4 (ISO)、小麦胚芽凝集素 (WGA)、番茄凝集素 (LYCO) 或 FITC 标记凝胶）。切除腓肠肌、伸肌、膈肌、臀大肌和绉肌，将其固定在最佳肌节长度处，并使用荧光显微镜观察：结果：WGA 和 LYCO 能有效标记整个血管网络，WGA 标记的毛细血管更亮。ISO 能标记动脉血管和毛细血管的早期部分，但不能标记整个毛细血管或静脉网络。FITC 标记的凝胶能有效标记微血管网，但并非所有小血管都能被一致标记。在测试的所有肌纤维类型中，每种标记方法的染色模式都相似：结论：WGA 是灌注标记和观察整装骨骼肌制备中完整微血管网络的最佳方法，可与 ISO 结合使用，以区分网络的动脉侧和静脉侧。

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来源期刊

Journal of Vascular Research 医学-生理学

CiteScore

3.40

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： The ''Journal of Vascular Research'' publishes original articles and reviews of scientific excellence in vascular and microvascular biology, physiology and pathophysiology. The scope of the journal covers a broad spectrum of vascular and lymphatic research, including vascular structure, vascular function, haemodynamics, mechanics, cell signalling, intercellular communication, growth and differentiation. JVR''s ''Vascular Update'' series regularly presents state-of-the-art reviews on hot topics in vascular biology. Manuscript processing times are, consistent with stringent review, kept as short as possible due to electronic submission. All articles are published online first, ensuring rapid publication. The ''Journal of Vascular Research'' is the official journal of the European Society for Microcirculation. A biennial prize is awarded to the authors of the best paper published in the journal over the previous two years, thus encouraging young scientists working in the exciting field of vascular biology to publish their findings.