White H D, Walker M L, Trinick J
Department of Biochemistry, Eastern Virginia Medical School, Norfolk 23507, USA.
J Struct Biol. 1998;121(3):306-13. doi: 10.1006/jsbi.1998.3968.
Apparatus is described for the kinetic investigation of biological reactions by electron cryomicroscopy with time resolution on the order of milliseconds. This involves layering a grid with one reactant and then spraying on a second reactant immediately before freezing. Two-stage mixing can be achieved by mixing two solutions, holding them in a delay line for a preset interval, and then spraying the aged solution onto a grid carrying a third reactant. The individual steps of these procedures are under software control and can be adjusted independently. Spray-freezing is widely applicable since solutions of small molecules, proteins, and protein assemblies can be delivered as aerosols. Thus the method can be used to study both the effects of small molecules on macromolecules and for monitoring protein-protein interactions. It may also be useful in other situations, for instance in light microscopy.
描述了一种通过电子冷冻显微镜对生物反应进行动力学研究的装置，其时间分辨率可达毫秒级。这包括在网格上铺上一种反应物，然后在冷冻前立即喷洒第二种反应物。通过将两种溶液混合，在延迟线中保持预设时间间隔，然后将老化后的溶液喷洒到带有第三种反应物的网格上，可以实现两级混合。这些程序的各个步骤均由软件控制，并且可以独立调整。喷雾冷冻具有广泛的适用性，因为小分子、蛋白质和蛋白质组装体的溶液都可以以气溶胶形式输送。因此，该方法可用于研究小分子对大分子的影响以及监测蛋白质-蛋白质相互作用。它在其他情况下也可能有用，例如在光学显微镜中。
