Mollenhauer H H, Totten C
J Cell Biol. 1971 Feb;48(2):387-94. doi: 10.1083/jcb.48.2.387.
Several fixation procedures were studied to determine those most suitable for preservation of seeds during late stages of development and early stages of germination. These are the periods when the tissues are partially dehydrated and are most difficult to fix for electron microscopy. It was found that a prefixation with a mixture of glutaraldehyde, reconstituted formaldehyde (i.e. paraformaldehyde), and acrolein, followed by a postfixation in OsO(4) or KMnO(4), gives very acceptable images. The results also indicate that glutaraldehyde is necessary for preservation of cell shape, paraformaldehyde for stabilization of reserve proteins, and acrolein for rapid penetration of tissues. Phosphate, cacodylate, and collidine are all acceptable buffers, although collidine gives the most consistent results.
研究了几种固定程序,以确定最适合在种子发育后期和萌发早期保存种子的方法。在这些时期,组织会部分脱水,并且最难为电子显微镜固定。结果发现,先用戊二醛、重构甲醛(即多聚甲醛)和丙烯醛的混合物进行预固定,然后用四氧化锇(OsO₄)或高锰酸钾(KMnO₄)进行后固定,能得到非常理想的图像。结果还表明,戊二醛对于保持细胞形状是必要的,多聚甲醛对于储备蛋白的稳定是必要的,而丙烯醛对于组织的快速渗透是必要的。磷酸盐、二甲胂酸盐和可力丁都是合适的缓冲剂,不过可力丁给出的结果最稳定。