Confining protein crystallization to a millimetre size was achieved within 0.5 h after stopping 1 h intense trapping laser irradiation, which shows excellent performance in spatial and temporal controllability compared to spontaneous nucleation. A continuous-wave near-infrared laser beam is tightly focused into a glass/solution interfacial layer of a supersaturated buffer solution of hen egg-white lysozyme (HEWL). The crystallization is not observed during laser trapping, but initiated by stopping the laser irradiation. The generated crystals are localized densely in a circular area with a diameter of a few millimetres around the focal spot and show specific directions of the optical axes of the HEWL crystals. To interpret this unique crystallization, we propose a mechanism that nucleation and the subsequent growth take place in a highly concentrated domain consisting of HEWL liquid-like clusters after turning off laser trapping.

We propose the dynamics and mechanism of laser trapping-induced crystal growth of hen egg-white lysozyme (HEWL). A continuous-wave near-infrared laser beam is used as a trapping light source and focused at a point 10 μm away from a target tetragonal HEWL crystal that is spontaneously generated in solution. Laser trapping of HEWL liquid-like clusters in solution increases local concentration in the focus, where the free motion and orientation of the clusters are strongly restricted, and the clusters show high rigidity and ordering. The cluster association and reorientation at the micrometer-sized focus is evolved to a large highly concentrated domain of the clusters, where the specific target crystal is grown. Initially, the high rigidity and ordering of the clusters strongly suppress the crystal grow rate compared to spontaneous crystal growth. Continuous laser trapping at the focus of the initially formed domain, however, leads to the transition to another domain with different concentration, rigidity, and ordering of the clusters, which surprisingly enhances the crystal growth rate. More interestingly, the clusters in both domains have anisotropic features reflecting the laser polarization direction, which also contributes to the crystal growth.

On page 16, left column, second paragraph, first line, of this paper, the notation of sample of HEWL (Wako, >99.0%) is incorrect. The correct notation is as follows: HEWL (Wako, for Biochemistry).