Microscope detection options for colorless protein crystals grown in lipidic cubic phases.
Microscope detection options for colorless protein crystals grown in lipidic cubic phases.
复制标题
用于在脂质立方相中生长的无色蛋白质晶体的显微镜检测选项。
DOI：
10.1107/s0021889803013724
复制
发表时间：
2003
影响因子：
6.1
通讯作者：
Nollert,Peter
中科院分区：
文献类型：
--
作者：
Nollert,Peter
使用无规立方相作为晶体成核和生长基质正变得流行，并且已经产生可溶性和膜蛋白质的晶体。到目前为止，通过这种方法结晶的所有膜蛋白都被着色。这一特点有助于在最初的筛选中检测经常遇到的微晶。实际上，由于蛋白质晶体和周围的无定形立方相的折射率的小差异，小的无色蛋白质晶体具有差的光学对比度。虽然一个完美的制备立方相是透明的和光学各向同性的，在结晶设置，它经常掩盖晶体由于裂纹，夹杂物，表面变形和相边界。在这里，几个专门的显微镜技术和照明条件进行了比较，它被发现，足够的对比度产生的交叉偏振显微镜和霍夫曼调制对比度显微镜检测无色蛋白质晶体。
The use of lipidic cubic phases as crystal nucleation and growth matrices is becoming popular and has yielded crystals of soluble and membrane proteins. So far, all of the membrane proteins crystallized by this method have been colored. This feature has facilitated the detection of the often encountered microcrystals in initial screening rounds. Indeed, small colorless protein crystals have poor optical contrast as a result of the small differences in refractive index of the protein crystal and the surrounding lipidic cubic phase. While a perfect preparation of a lipidic cubic phase is transparent and optically isotropic, in a crystallization setup it frequently disguises crystals due to cracks, inclusions, surface distortions and phase boundaries. Here, several specialized microscopic techniques and illumination conditions are compared and it is found that sufficient contrast is generated by cross polarization microscopy and by Hoffman modulation contrast microscopy for the detection of colorless protein crystals.