Gounon P, Rolland J P
Institut Pasteur, Station Centrale de Microscopie Electronique, Paris, France.
Micron. 1998 Aug;29(4):293-6. doi: 10.1016/s0968-4328(98)00013-4.
We introduced a slight modification of Unicryl for fast polymerization at low temperature (in the range-20 degree C to-35 degree C) without modification of both physical properties and immunocytochemical yield. An UV initiator benzoin ethyl ether (BEE) is added at 0.05% concentration in neat resin during infiltration and final polymerization at-35 degree C. Immunogold labeling of amylase of mouse pancreas embedded in standard conditions recommended by the manufacturer or with our modification were compared by counting colloidal gold particles over zymogen granules using MACS, a specialized software dedicated to this operation. We noticed that addition of 0.05% BEE in Unicryl reduces the curing time to 16 h, which can be extended up to a maximum of 48 h at-35 degree C without noticeable modification of ultrastructural morphology, physical properties and immunocytochemical yield.
我们对Unicryl进行了轻微改良,使其能在低温(-20摄氏度至-35摄氏度)下快速聚合,同时不改变其物理性质和免疫细胞化学产率。在渗透过程中,于纯树脂中添加浓度为0.05%的紫外线引发剂安息香乙醚(BEE),并在-35摄氏度下进行最终聚合。通过使用专门用于此操作的软件MACS,对包埋在制造商推荐的标准条件下或经我们改良的条件下的小鼠胰腺淀粉酶进行免疫金标记,通过计算酶原颗粒上的胶体金颗粒来进行比较。我们注意到,在Unicryl中添加0.05%的BEE可将固化时间缩短至16小时,在-35摄氏度下最长可延长至48小时,而不会对超微结构形态、物理性质和免疫细胞化学产率产生明显影响。