COUNTING OF MONO- AND BINUCLEATED CELLS IN TISSUE SECTIONS
COUNTING OF MONO- AND BINUCLEATED CELLS IN TISSUE SECTIONS
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DOI：
10.1002/ar.1091420402
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发表时间：
1962-01-01
影响因子：
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通讯作者：
PATTERSON, D
中科院分区：
文献类型：
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作者：
CARRIERE, R;PATTERSON, D
对组织切片中的双核细胞进行计数可能会出现错误，因为切片可能会导致它们看起来是单核的，并且带有切片细胞核的单核或双核细胞可能会在两个连续切片的每一个中看到。使用大鼠肝脏研究了这一技术问题，其中单核细胞和双核细胞都可以含有二倍体或多倍体细胞核。材料的初始制备和组织计量程序必须严格标准化，以尽量减少各个切片之间的细胞学差异。推导出校正公式；第一个与 Pfuhl (30) 提出的相同，但确定其常数的程序不同，并且仅校正了双核细胞的数量；通过第二个公式处理单核细胞的数量。公式中使用的常数之一是 2q 或平均截面厚度。实际上，这总是比切片机设置指示的要大一些，并且因子 2q 可能会在获得的结果中产生小误差。然而，每个实验组有 6-8 个切片，通过仔细的切片技术可以将这种误差减少到几乎可以忽略不计的比例。测量核大小以获得所使用的两个常数，并在计数细胞时对不同倍性的核进行分类。描述了这些测量的程序，并说明了概率坐标纸上核尺寸分布的分析。值得注意的是，组织切片中二倍体和四倍体大小的细胞核的相对数量的测定是可靠的，通常包括2-3%量级的组织计量误差。
Counting binucleated cells in histological sections is subject to error because sectioning can cause them to appear mononucleated, and because mono- or binucleated cells with sectioned nuclei may be seen in each of two consecutive sections. This technical problem was investigated using rat liver, where both mono- and binucleated cells can contain diploid or polyploid nuclei. The initial preparation of the material and the histometric procedures must be rigidly standardized to minimize cytological variations among individual sections. Correction formulae were derived; the first was identical to that proposed by Pfuhl (30), but the procedure for determining its constants differed, and this corrected the numbers of binucleated cells only; the numbers of mononucleated cells were treated by means of a second formula. One of the constants used in the formulae is 2q or average section thickness. In practice, this is always somewhat greater than indicated by the microtome setting, and the factor 2q can produce a small error in the results obtained. However, with 6-8 sections per experimental group, this error can be reduced to near negligible proportions by careful sectioning technique. Nuclear sizes were measured to obtain two of the constants used, and to classify nuclei of different ploidy while counting cells. The procedure for these measurements was described, and the analysis of nuclear size distributions on probability graph paper was illustrated. It was noted that the determination of the relative numbers of diploid- and tetraploid-sized nuclei in tissue sections is reliable, usually comprising a histometric error of the order of 2-3%.