Change Details

= 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) {F10627035} - 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) as you would want them - Do not use averaging or Z stacks === Prepare for acquisition === - Check the {nav Regions} tool in ZEN and create a rectangle in a region of your image that contains signal - In the {nav Regions} tool, uncheck **Bleach**, but keep **Acquisition** & **Analysis** - Check **Fit frame size to bounding rectangle of resions** - Check {nav Time Series} - Use 100 timepoints (or more if you want) === Run Acquisition === - Use {nav Start Experiment } - Switch to the **Mean ROI** tab on the left of the image === Copy the data and run the macro === - On the table, {nav right click} and select **Copy Data** - Open Fiji and open the macro `Assess Bleaching.ijm` (Drag & Drop the file into Fiji) {F10627035} - Run the macro. It will use the data that is currently on the Windows clipboard and return the results === See the results === {F10627023, size=full} There will be a 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) readyas you would want them - Do not use averaging or Z stacks === Prepare for acquisition === - Using- Check the "{nav Regions"} tool in ZEN, and create a rectangle in a region of your image withthat contains signal - In the {nav Regions menu} tool, uncheck "**Bleach"**, but keep "**Acquisition" & "** & **Analysis"** - Check "Use**Fit frame size to bounding rectangle blablabla"of resions** - Check "{nav Time Series"} - Use 100 timepoints (or more if you want) {F10627033} TODO, illustrate image better === Run Acquisition === - Use {nav Start Experiment } - Switch to the "**Mean ROI"** tab on the left of the image === Copy the data and run the macro === - On the table, {nav right click} and select "**Copy Data"** - Open Fiji and open the macro "`Assess Bleaching.ijm"` (Drag & Drop the file into Fiji) {F10627035} - Run the macro. It will use the data that is currently on the Windows clipboard and return the results === See the results === {F10627023, size=full} MThere will be a 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.