Apgar J M, Juarranz A, Espada J, Villanueva A, Cañete M, Stockert J C
Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Cantoblanco, Spain.
J Microsc. 1998 Jul;191(Pt 1):20-7. doi: 10.1046/j.1365-2818.1998.00348.x.
The fluorescence pattern induced by haematoxylin-eosin (HE) and Masson's trichrome (MT) staining methods on paraffin sections of rat embryos (from 13 to 18 days old) has been studied. Using optimal excitation (green light, 545 nm), HE- or MT-stained sections showed a selective red emission of the acidophilic tissue components, which was due to eosin Y in the case of HE and to acid fuchsin and/or xylidine ponceau in the case of MT. The fluorescence intensity induced by these anionic dyes was variable and related to the substrate nature and the embryo age. Whereas in young embryos only the immature red blood cells showed a noticeable fluorescence, in the oldest embryos there were also other tissue components that selectively fluoresced with these dyes, in particular fibre lens cells, elastic fibres, zymogen granules and muscle cells. Spectrofluorometric studies on free dyes and densitometric analysis of protein blots confirmed microscopical observations. Our results indicate that the standard HE and MT staining methods can be used in recognizing the appearance of specific protein structures in embryonic tissues by means of fluorescence microscopy.
研究了苏木精-伊红（HE）和马松三色（MT）染色方法对大鼠胚胎（13至18日龄）石蜡切片诱导的荧光模式。使用最佳激发光（绿光，545nm），HE或MT染色切片显示嗜酸性组织成分有选择性的红色发射，HE染色情况下是由于伊红Y，MT染色情况下是由于酸性品红和/或二甲苯红紫。这些阴离子染料诱导的荧光强度是可变的，并且与底物性质和胚胎年龄有关。在年轻胚胎中，只有未成熟红细胞显示出明显的荧光，而在最老的胚胎中，也有其他组织成分用这些染料选择性地发出荧光，特别是晶状体纤维细胞、弹性纤维、酶原颗粒和肌肉细胞。对游离染料的荧光光谱研究和蛋白质印迹的光密度分析证实了显微镜观察结果。我们的结果表明，标准的HE和MT染色方法可用于通过荧光显微镜识别胚胎组织中特定蛋白质结构的外观。
