Cassell M D, Mankovich N J, Gray T S, Williams T H
Peptides. 1982 May-Jun;3(3):283-90. doi: 10.1016/0196-9781(82)90088-2.
When viewed under dark-field illumination, peptidergic terminals in sections stained by the Sternberger PAP immunocytochemical method are seen as individual points of light. Under high magnification, the degree of brightness of various areas of immunoreactive terminals is seen to be a function of the density of terminals in these areas. By analyzying the relative brightness of the immunostained central nucleus of the amygdala (CNA) with an EyeCom II PDP-11/34 image analysis system, we have obtained a relative evaluation of the density distribution of neurotensin (NT)-, substance P (SP), VIP-, angiotensin II (AII), m-enkephalin (m-ENK) and somatostatin (SS)-immunoreactive terminals in terms of normal morphology and following a brain lesion. The EyeCom II system divides the presented image into 307200 picture elements (pixels) and assigns one of 256 grey values to the average brightness with each pixel. We have aggregated the grey level frequencies into 5 levels where level 1 corresponds to the highest terminal density and level 5 to the lowest density. At level 1, only NT- and VIP-immunoreactive terminals occupy a significant percentage of the cross-sectional area of the CNA (20%). About 15% of the area of the CNA has VIP terminals with level 5 density. The distributions of the top 20% of the terminal density range of NT, SP, AII and VIP support a classical medial/lateral division of the nucleus. The distribution of the same range of SS- and ENK terminals suggests a dorsoventral division of the CNA. A preliminary study indicates that comparison of grey level frequency histograms generated by image analysis from homologous lesioned and unlesioned sections of the CNA can yield useful information regarding post-lesion changes in the distribution of immunoreactive terminals.
在暗视野照明下观察，用Sternberger PAP免疫细胞化学方法染色的切片中的肽能终末呈现为一个个亮点。在高倍放大下，免疫反应性终末不同区域的亮度程度被视为这些区域终末密度的函数。通过使用EyeCom II PDP - 11/34图像分析系统分析杏仁核中央核（CNA）免疫染色的相对亮度，我们在正常形态以及脑损伤后，对神经降压素（NT）、P物质（SP）、血管活性肠肽（VIP）、血管紧张素II（AII）、甲硫氨酸脑啡肽（m - ENK）和生长抑素（SS）免疫反应性终末的密度分布进行了相对评估。EyeCom II系统将呈现的图像划分为307200个图像元素（像素），并为每个像素的平均亮度分配256个灰度值之一。我们将灰度级频率汇总为5个级别，其中1级对应最高的终末密度，5级对应最低密度。在1级，只有NT和VIP免疫反应性终末占据CNA横截面积的显著百分比（20%）。CNA区域约15%有5级密度的VIP终末。NT、SP、AII和VIP终末密度范围前20%的分布支持该核的经典内侧/外侧划分。SS和ENK终末相同范围的分布表明CNA存在背腹侧划分。一项初步研究表明，通过图像分析从CNA同源损伤和未损伤切片生成的灰度级频率直方图进行比较，可以获得有关损伤后免疫反应性终末分布变化的有用信息。
