Zeiss LSM 880 Operating Manual page 494

ZEISS Center Screen Area / Image Containers - Display and Image Analysis
Example: FRAP Performed in a Nucleus Expressing GFP Labeled Proteins
Display of the image series in the Mean ROI display mode: The drawing tools are used to define the ROI
to be analyzed (ROI 1), the background ROI (ROI 3), and the reference ROI (ROI 2) (see Fig. 680). The
reference ROI must be a neighboring cell which has been imaged with the same laser intensity over time
identical to the cell, which has been bleached to induce FRAP. Make sure the whole cell or cell
compartment of interest is imaged and therefore illuminated.
Use the time slider in the Dimensions view options control block of the general view options
group to display the first image after the bleach event. This makes it easy to choose the ROI for
analysis. The analysis region should be slightly smaller than the region that has been bleached.
The latter is listed in the Mean of ROIs list in the graphics view options control block.
• Define your ROIs for analysis in the Mean of ROI View tab.
Fig. 680 FRAP data displaying the first bleached image of an images series in the Mean of ROI View
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CHAPTER 1 - SYSTEM OPERATION
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LSM 880
10/2014 V_01