| Imaging and Measurement of Nanomechanical Properties within Primary Xylem Cell Walls of Broadleaves
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Author:
Date:
2014-12-20
[Abstract] A technique of atomic force microscopy (AFM) called PeakForce quantitative nanomechanical mapping (PeakForce QNM) is an efficient tool for the quantitative mechanobiological imaging of fibrillar aggregate, human epidermal cell and woody plant cell wall topography (Sweers et al., 2011; Heu et al., 2012; Ďurkovič et al., 2012; Ďurkovič et al., 2013). Here, we describe a detailed protocol for the measurement of nanomechanical properties of primary xylem cell walls in woody plants, for the determination of reduced Young’s modulus of elasticity (MOE), adhesion, deformation, and energy dissipation (Figure 1). This new technique provides direct control of the maximum loading force and the deformation depth in cell wall samples keeping indentations small, while ...
[摘要] 称为PeakForce定量纳米机械映射（PeakForce QNM）的原子力显微镜（AFM）技术是用于纤维聚集体，人表皮细胞和木本植物细胞壁形貌的定量机械生物学成像的有效工具（Sweers等人， 2011; Heu等人，2012;Ďurkovič等人，2012;Ďurkovič等人，2013年）。在这里，我们描述一个详细的协议，用于测量木本植物中主要木质部细胞壁的纳米机械性能，用于确定降低的杨氏弹性模量（MOE），粘附，变形和能量耗散（图1）。这种新技术提供了对细胞壁样品中的最大负载力和变形深度的直接控制，保持压痕小，同时消除损伤侧向力，以便保持AFM尖端和植物样品。高分辨率和非破坏性成像为木质植物细胞壁的结构生物学提供了新的定量机械的见解。该程序还可适用于具有变化的刚度范围的其他生物样品。
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| Maize Kernels – Fixation in FAA, Embedding, Sectioning and Feulgen Staining
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Author:
Date:
2013-08-05
[Abstract] The protocol describes preparation of young developing maize kernels for microscopical analysis of nuclei in tissue sections. The fixative FAA (formaldehyde, acetic acid and ethanol) is suitable for preservation of nuclear morphology and allows for quantitative staining of DNA with Schiff reagent in Feulgen staining. The fixation and embedding protocol may be used also for various other histology staining procedures, but care must be taken as the cytoplasm usually shrinks a bit using this procedure. The protocol was used for analysis of seed development of various maize lines, mutants and maize relatives (Vilhar et al., 2002; Kladnik et al., 2006; Dermastia et al., 2009; Bernardi et al., 2012).
[摘要] 该协议描述了用于组织切片中核的显微镜分析的年轻发育的玉米籽粒的制备。 固定剂FAA（甲醛，乙酸和乙醇）适合保存核形态，并允许在Feulgen染色中用希夫试剂定量染色DNA。 固定和包埋方案也可以用于各种其他组织学染色程序，但是必须小心，因为细胞质通常使用该程序略微收缩。 该方案用于分析各种玉米品系，突变体和玉米亲缘种的种子发育（Vilhar等人，2002; Kladnik等人，2006; Dermastia >等人，2009; Bernardi等人，2012）。
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