PT - JOURNAL ARTICLE AU - Karen L. Anderson AU - Christopher Page AU - Mark F. Swift AU - Dorit Hanein AU - Niels Volkmann TI - Accurate alignment between correlated light, cryo-light, and electron cryo-microscopy data using sample support features AID - 10.1101/099077 DP - 2017 Jan 01 TA - bioRxiv PG - 099077 4099 - http://biorxiv.org/content/early/2017/01/07/099077.short 4100 - http://biorxiv.org/content/early/2017/01/07/099077.full AB - Combining fluorescence microscopy with electron cryo-tomography allows, in principle, spatial localization of tagged macromolecular assemblies and structural features within the cellular environment. To allow precise localization and scale integration between the two disparate imaging modalities, accurate alignment procedures are needed. Here, we describe a marker-free method for aligning images from light or cryo-light fluorescence microscopy and from electron cryo-microscopy that takes advantage of sample support features, namely the holes in the carbon film. We find that the accuracy of this method, as judged by prediction errors of the hole center coordinates, is better than 100 nm.