Pegg D E, Diaper M P, Scholey S E, Coombs R R
J Immunol Methods. 1982 Nov 12;54(3):331-41. doi: 10.1016/0022-1759(82)90317-9.
Red cells, trypsin-treated to render them more agglutinable and coupled with antiglobulin reagents, may be preserved by droplet freezing in liquid nitrogen. A 2% cell suspension in 0.45% w/v sodium chloride, 5% w/v sucrose and 10% w/v dextran 40 was used. After thawing the frozen pellets in phosphate-buffered saline at 40 degrees C, more than 80% cell recovery was obtained. Sheep and ox red cells preserved in this way were as satisfactory in antiglobulin and in reverse passive haemagglutination tests as unfrozen indicator red cells. Trypsin-treated human red cells coupled with anti-IgE could likewise be frozen and on reconstitution used to assay IgE in human serum. Reconstituted ox red cells were slightly less efficient in rosetting than cells which had not been frozen.
经胰蛋白酶处理使其更易于凝集并与抗球蛋白试剂结合的红细胞,可通过液氮滴冻法保存。使用的是在0.45%(w/v)氯化钠、5%(w/v)蔗糖和10%(w/v)右旋糖酐40中的2%细胞悬液。在40℃的磷酸盐缓冲盐水中解冻冷冻的小丸后,细胞回收率超过80%。以这种方式保存的绵羊和牛红细胞在抗球蛋白试验和反向被动血凝试验中与未冷冻的指示红细胞一样令人满意。经胰蛋白酶处理并与抗IgE结合的人红细胞同样可以冷冻,复溶后用于检测人血清中的IgE。复溶后的牛红细胞在形成玫瑰花结方面的效率略低于未冷冻的细胞。