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Abstract：
Assessing fish liver status is common in aquaculture nutrition assays. This often implies determining hepatocytes profile areas in routine thin (5-7 μm) histological sections. However, there are theoretical problems using planar morphometry in thin sections: inherent sampling cells biases, too small numbers of sampled cells, under/overestimation of size, measuring size as areas when cells are three-dimensional (3D) entities. The gold standard for assessing/validate cell size is stereology using thick sections (20-40 μm). Here, we estimated the volume of hepatocytes and their nuclei by the nucleator and optical disector stereological probes (in thick sections), and, innovatively, in thin sections too (using single-section disectors). The liver of common carp eating feed containing either low or high level of lipids was targeted. Results were compared with prior profile areas from planar morphometry using thin sections, and with profile areas estimated here with the two-dimensional (2D) nucleator. Ratios between nucleus and cell/cytoplasm (N/C) areas and volumes were calculated and compared. There was high positive correlation between volumes in thin and thick sections (r = .85 to .89; p < .001), empirically validating the single-section disector. Strong correlations existed between profile-derived versus 2D-nucleator areas (r = .74 to .83; p < .001). There was systematic underestimation of cells and nucleus size using planar morphometry. The N/C ratios derived from the 2D-nucleator data were higher than those from planar morphometry. Despite theoretical premises for using simple planar morphometry in thin sections are flawed, our results support that such morphometry on carp/fish hepatocytes may offer some valid biological conclusions. Anyway, we advanced guidelines for implementing proper methods.
摘要：
评估鱼肝状态在水产养殖营养测定中很常见。这通常意味着确定常规薄（5-7μm）组织学切片中的肝细胞分布区域。然而,在薄切片中使用平面形态测量法存在理论问题：固有的采样细胞偏差，采样的细胞数量太少了，低估/高估尺寸，当细胞是三维(3D)实体时，测量大小作为面积。评估/验证细胞大小的黄金标准是使用厚切片(20-40μm)的立体学。这里,我们通过成核剂和光学解剖体视学探针（厚切片）估计肝细胞及其细胞核的体积，and,创新,在薄切片中也是如此(使用单切片切片器)。目标是食用含有低或高脂质水平的鲤鱼饲料的肝脏。使用薄切片将结果与平面形态测量的先前轮廓区域进行比较，并在此处使用二维（2D）成核器估算轮廓面积。计算并比较细胞核与细胞/细胞质(N/C)面积和体积之间的比率。薄切片和厚切片的体积之间存在高度正相关(r=.85至.89；p<.001)，对单节数据进行了实证验证。轮廓衍生面积与2D成核面积之间存在很强的相关性（r=.74至.83；p<.001）。使用平面形态测量法系统地低估了细胞和细胞核的大小。从2D成核剂数据得出的N/C比高于平面形态计量学得出的N/C比。尽管在薄切片中使用简单平面形态计量学的理论前提存在缺陷，我们的结果支持，这种形态计量学对鲤鱼/鱼肝细胞可能提供一些有效的生物学结论。不管怎样,我们提出了实施适当方法的指导方针。
