PT - JOURNAL ARTICLE AU - Herbert J. Bernstein AU - Lawrence C. Andrews AU - James Foadi AU - Martin R. Fuchs AU - Jean Jakoncic AU - Sean McSweeney AU - Dieter K. Schneider AU - Wuxian Shi AU - John Skinner AU - Alexei Soares AU - Yusuke Yamada TI - Serial Crystallography with Multi-stage Merging oi 1000’s of Images AID - 10.1101/141770 DP - 2017 Jan 01 TA - bioRxiv PG - 141770 4099 - http://biorxiv.org/content/early/2017/05/25/141770.short 4100 - http://biorxiv.org/content/early/2017/05/25/141770.full AB - KAMO and Blend provide particularly effective tools to automatically manage the merging of large numbers of data sets from serial crystallography. The requirement for manual intervention in the process can be reduced by extending Blend to support additional clustering options to increase the sensitivity to differences in unit cell parameters and to allow for clustering of nearly complete datasets on the basis of intensity or amplitude differences. If datasets are already sufficiently complete to permit it, apply KAMO once, just for reflections. If starting from incomplete datasets, one applies KAMO twice, first using cell parameters. In this step either the simple cell vector distance of the original Blend is used, or the more sensitive NCDist, to find clusters to merge to achieve sufficient completeness to allow intensities or amplitudes to be compared. One then uses KAMO again using the correlation between the reflections at the common HKLs to merge clusters in a way sensitive to structural differences that may not perturb the cell parameters sufficiently to make meaningful clusters.Many groups have developed effective clustering algorithms that use a measurable physical parameter from each diffraction still or wedge to cluster the data into categories which can then be merged to, hopefully, yield the electron density from a single protein iso-form. What is striking about many of these physical parameters is that they are largely independent from one another. Consequently, it should be possible to greatly improve the efficacy of data clustering software by using a multi-stage partitioning strategy. Here, we have demonstrated one possible approach to multi-stage data clustering. Our strategy was to use unit-cell clustering until merged data was of sufficient completeness to then use intensity based clustering. We have demonstrated that, using this strategy, we were able to accurately cluster data sets from crystals that had subtle differences.