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. Author manuscript; available in PMC: 2015 Mar 15.
Published in final edited form as: Methods. 2013 Aug 28;66(2):139–152. doi: 10.1016/j.ymeth.2013.08.017

Figure 1. Schematic representation of background substraction, SBT correction and ROI selection FRET analysis protocol.

Figure 1

Seven steps are required for the FRET analysis protocol: 1) Zeiss format lsm image opening, 2) background removal, 3) SBT correction (PFRET software), 4) ROI selection, 5) overlapping ROI removal, 6) Extract and processing of FRET related ROI data, and 7) plot charts and perform statistical analysis. Three of those steps in the Image J ROI selection macro are described in detail, including background removal (panel a), ROI selection (panel b) and removal of overlapping ROIs (panel c).