= 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 ===

= 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) {F10627033} TODO, illustrate image better === 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 === {F10627023, size=full} Message with number of frames before 50% of intensity is lost. This is also reflected in a new line in a Table called Bleach Assessment Finally, you can observe the plot to see if the exponential fit worked

= 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) {F10627033} TODO, illustrate image better === 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 === {F10627023, size=full} Message with number of frames before 50% of intensity is lost. This is also reflected in a new line in a Table called Bleach Assessment Finally, you can observe the plot to see if the exponential fit worked