= Quickly Assessing Fluorescence Bleaching in LSM700 Microscopes with ZEN and Fiji= == Protocol == === Initial Settings === - On the LSM700, make sure you have all settings (Laser, Gain, Pixel Size, Speed) ready - Do not use averaging or Z stacks === Prepare for acquisition === - Using the "Regions" tool in ZEN, create a rectangle in a region of your image with signal - In the Regions menu, uncheck "Bleach", but keep "Acquisition" & "Analysis" - Check "Use bounding rectangle blablabla" - Check "Time Series" - Use 100 timepoints (or more if you want) === Run Acquisition === - Use Start Experiment - Switch to the "Mean ROI" tab on the left of the image === Copy the data and run the macro === - On the table, right click and select "Copy Data" - Open Fiji and open the macro "Assess Bleaching.ijm" (Drag & Drop the file into Fiji) - Run the macro. It will use the data that is currently on the Windows clipboard === See the results ===

Document migrated to https://wiki-biop.epfl.ch/en/Teaching/PCB/assess-bleach