Amenomori T, Tomonaga M, Matsuo T, Yoshida Y, Kuriyama K, Sadamori N, Ichimaru M
Int J Cell Cloning. 1985 May;3(3):133-42. doi: 10.1002/stem.5530030301.
We report a micromethod for chromosome preparation from individual hematopoietic colonies cultured in methylcellulose. The entire process was carried out on poly-Lysine (PL)-coated slides. Individual colonies were transferred into 10 microliter of 0.075 M KCl and placed on PL-coated slides. After hypotonic treatment of the colony cells and their attachment to the slides, the cells were fixed by a three-step procedure as follows: addition of a 30% fixative (3:1 methanol:acetic acid) diluted with the hypotonic solution, addition of 20% ethanol, and subsequent immersion of the slides in a 100% fixative. The slides were flame dried and Giemsa stained. Q- and G-banding techniques also were used. These procedures provided analyzable chromosome preparations, even from colonies containing fewer than 50 cells.
我们报告了一种从在甲基纤维素中培养的单个造血集落制备染色体的微量方法。整个过程在涂有聚赖氨酸（PL）的载玻片上进行。将单个集落转移到10微升0.075 M KCl中，并置于涂有PL的载玻片上。在对集落细胞进行低渗处理并使其附着在载玻片上后，通过以下三步程序固定细胞：加入用低渗溶液稀释的30%固定剂（3:1甲醇：乙酸），加入20%乙醇，随后将载玻片浸入100%固定剂中。将载玻片火焰干燥并进行吉姆萨染色。还使用了Q带和G带技术。即使是来自细胞少于50个的集落，这些程序也能提供可分析的染色体标本。
