Leif R C, Ingram D, Clay C, Bobbitt D, Gaddis R, Leif S B, Nordqvist S
J Histochem Cytochem. 1977 Jul;25(7):538-43. doi: 10.1177/25.7.894002.
In order to monitor the development of a cell dissociation technique, it was essential to utilize the Centrifugal Cytology rotor to produce glutaraldehyde-fixed even cellular dispersions. The Cytology rotor has been improved to insure rapid alignment with the centrifugal field during both acceleration and deceleration, and the fixative is now delivered to the surface of the slide. The dissociation of the cells results in a loss of their adhesion to glass slides. Three bonding agents were tested: (a) Poly-L-Lysine; (b) Mayer's albumin fixative; (c) positively charging the slides with a silicone coating. The results with 65% albumin-coated slides were clearly superior to the other two. The addition of a postfixation step of 95% ethanol/4% polyethylene glycol did not significantly affect the recovery of the cells, but did eliminate some unevenness in the Centrifugal Cytology preparations, flattened the cells and expedited the procedure.
为了监测细胞解离技术的发展,利用离心细胞学转子来制备戊二醛固定的均匀细胞悬液至关重要。细胞学转子已得到改进,以确保在加速和减速过程中都能快速与离心场对齐,并且现在固定剂被输送到载玻片表面。细胞解离导致它们与载玻片的附着力丧失。测试了三种粘合剂:(a) 聚-L-赖氨酸;(b) 迈耶氏白蛋白固定剂;(c) 用硅酮涂层对载玻片进行正电荷处理。65%白蛋白包被的载玻片的结果明显优于其他两种。添加95%乙醇/4%聚乙二醇的后固定步骤对细胞回收率没有显著影响,但确实消除了离心细胞学制备中的一些不均匀性,使细胞变平并加快了操作过程。