Three-Dimensional Photoactivated Localization Microscopy with Genetically Expressed Probes

We describe methods for converting a 2D PALM into a system capable of acquiring such 3D images, with a particular emphasis on instrumentation that is compatible with choosing relatively dim, genetically expressed photoactivatable fluorescent proteins (PA-FPs) as PALM probes. After reviewing the basics of 2D PALM, we detail astigmatic and multiphoton imaging approaches well suited to working with PA-FPs. We also discuss the use of open-source localization software appropriate for 3D PALM.

Source: Springer protocols feed by Imaging/Radiology - November 13, 2014 Category: Radiology Source Type: news