González Santander R, Martínez Cuadrado G, González-Santander Martínez M, Monteagudo M, Martínez Alonso F J, Toledo Lobo M V
Department of Morphological Sciences and Surgery, Faculty of Medicine, University of Alcalá de Henares, Madrid, Spain.
Microsc Res Tech. 1997 Feb 1;36(3):151-8. doi: 10.1002/(SICI)1097-0029(19970201)36:3<151::AID-JEMT2>3.0.CO;2-S.
The effects of different fixatives, dehydration procedures, and embedding media on the structural and ultrastructural preservation of young chick embryos (Hamburger and Hamilton stages 18-24) have been studied by means of light and electron microscopy techniques. Under the light microscope, the results obtained with the use of Bouin, glutaraldehyde, or glutaraldehyde-paraformaldehyde mixtures, followed by partial dehydration of the samples and the embedding with two different polar resins (Historesin and Unicryl), were compared with the results obtained using conventional paraffin-embedding methods. Cell and tissue shrinkage was determined by comparing blood cells from those embryos embedded in either of resins with those embedded in paraffin. Samples were also compared with blood smears, either methanol-fixed or unfixed, obtained from embryos at the same Hamburger and Hamilton stages. The results obtained when Unicryl and Araldite were used for electron microscopy have also been compared. When ultrastructural images from glutaraldehyde-tannic acid/osmium tetroxide fixed, Unicryl embedded samples were compared with those from araldite embedded samples, the same good results were observed with either of the resins. Araldite embedding requires a complete dehydration of the samples, while Unicryl allows the embedding of partially dehydrated embryos with optimal ultrastructural results. We suggest that these polar resins can be considered as complementary tools for embedding delicate embryonic tissues, allowing partial dehydration of the specimens with an excellent cell and tissue preservation.
利用光学显微镜和电子显微镜技术,研究了不同固定剂、脱水程序和包埋介质对雏鸡胚胎(汉伯格和汉密尔顿第18 - 24阶段)结构和超微结构保存的影响。在光学显微镜下,将使用波因氏液、戊二醛或戊二醛 - 多聚甲醛混合物,随后对样品进行部分脱水并使用两种不同极性树脂(Historesin和Unicryl)包埋所获得的结果,与使用传统石蜡包埋方法获得的结果进行比较。通过比较用这两种树脂包埋的胚胎中的血细胞与石蜡包埋的胚胎中的血细胞,来确定细胞和组织收缩情况。还将样品与来自相同汉伯格和汉密尔顿阶段胚胎的甲醇固定或未固定的血涂片进行比较。也比较了使用Unicryl和Araldite进行电子显微镜观察时获得的结果。当将戊二醛 - 单宁酸/四氧化锇固定、Unicryl包埋的样品的超微结构图像与Araldite包埋的样品的超微结构图像进行比较时,两种树脂都观察到了同样良好的结果。Araldite包埋需要对样品进行完全脱水,而Unicryl允许对部分脱水的胚胎进行包埋,并获得最佳的超微结构结果。我们建议,这些极性树脂可被视为包埋脆弱胚胎组织的补充工具,允许对标本进行部分脱水,同时能出色地保存细胞和组织。