摘要
Data collection with microcrystals at synchrotron radiation facilities is challenging because it is difficult to harvest and locate microcrystals. Moreover,microcrystals are sensitive to radiation damage; thus, typically, a complete data set cannot be obtained with a single microcrystal. Herein, we report a new method for data collection with multiple microcrystals having a crystal size ranging from 1 to 30 lm. This method is suitable for not only low-temperature(100 K) data collection but also room-temperature data collection. Thin Kapton membranes were used to capture multiple crystals simultaneously. The microcrystals were visible under an optical microscope and easily located because the membrane was transparent and sufficiently thin. The film was fixed to a bracket that was prepared using a three-dimensional printer. The bracket was fixed on a magnetic base via screwing and employed by the goniometer system for data collection. Multiple data sets of fatty acid-binding protein 4(FABP4) and lysozyme microcrystals were collected using this novel designed device. Finally, the structures of protein FABP4 and lysozyme were obtained from these data via the molecule replacement method. The data statistics reveal that this method provides a comparable result to traditional methods such as a nylon loop.
Data collection with microcrystals at synchrotron radiation facilities is challenging because it is difficult to harvest and locate microcrystals. Moreover,microcrystals are sensitive to radiation damage; thus, typically, a complete data set cannot be obtained with a single microcrystal. Herein, we report a new method for data collection with multiple microcrystals having a crystal size ranging from 1 to 30 lm. This method is suitable for not only low-temperature(100 K) data collection but also room-temperature data collection. Thin Kapton membranes were used to capture multiple crystals simultaneously. The microcrystals were visible under an optical microscope and easily located because the membrane was transparent and sufficiently thin. The film was fixed to a bracket that was prepared using a three-dimensional printer. The bracket was fixed on a magnetic base via screwing and employed by the goniometer system for data collection. Multiple data sets of fatty acid-binding protein 4(FABP4) and lysozyme microcrystals were collected using this novel designed device. Finally, the structures of protein FABP4 and lysozyme were obtained from these data via the molecule replacement method. The data statistics reveal that this method provides a comparable result to traditional methods such as a nylon loop.
基金
supported by the Strategic Priority Research program of the Chinese Academy of Sciences(No.XDB08030101)
