Goldberg Stanley
Glen Mills Inc., 220 Delawanna Avenue, Clifton, NJ, 07014, USA,
Methods Mol Biol. 2015;1295:1-20. doi: 10.1007/978-1-4939-2550-6_1.
This chapter covers the various methods of Mechanical Cell Disruption and Tissue Homogenization that are currently commercially available for processing minute samples (<1 mL) to larger production quantities. These mechanical methods of lysing do not introduce chemicals or enzymes to the system. However, the energies needed when using these "harsh" methods can be high and destroy the very proteins being sought.The destruction of cell membranes and walls by these "harsh" methods is effected by subjecting the cells (1) to shearing by liquid flow, (2) to exploding by pressure differences between inside and outside of cell, (3) to collision forces by impact of beads or paddles, or (4) a combination of these forces. Practical suggestions to optimize each method, where to acquire such equipment, and links to reference sources are included.
本章介绍了目前市面上可用于处理微量样本(<1 mL)至更大产量的各种机械细胞破碎和组织匀浆方法。这些机械裂解方法不会向系统中引入化学物质或酶。然而,使用这些“严苛”方法所需的能量可能很高,会破坏正在寻找的蛋白质。这些“严苛”方法对细胞膜和细胞壁的破坏是通过使细胞(1)受到液体流动的剪切作用,(2)受到细胞内外压力差导致的爆裂作用,(3)受到珠子或桨叶撞击产生的碰撞力作用,或(4)这些力的组合来实现的。文中还包括了优化每种方法的实用建议、获取此类设备的途径以及参考资料来源链接。