Transfection_Efficiency_Tool
The tool helps to measure the transfection efficiency. It reports the percentage of transfected cells in the image. It has tools to manually correct the segmented nuclei (merge and split).
The source code in git-hub can be found here.
To install the tool save the file transfection_efficiency_tool.ijm into the folder macros/toolsets of your FIJI installation.
Select the "transfection_efficiency_tool" toolset from the >> button of the ImageJ launcher.
- The first button (the one with the image) opens this help page
- Press the d-button to detect the nuclei
- Press the m-button to activate the merge-tool for manual editing
- Make a line selection and press the s-button to split a nucleus
- Press the a-button to get the sorted intensity measurements of all nuclei
- Select a line in the results table and press the b-button to highlight the nuclei with a mean intensity above the selected one
- Press the c-button to count and report the transfected nuclei
Select the channel of your image, that contains the DAPI-staining of the nuclei. Press the d-button. You are asked to manually adjust the threshold. When done, press the ok-button on the *Action Required"-dialog.
- You can select the auto-thresholding method used before manual correction, when the threshold-tool is opened
Press the m-button to activate the merge tool. Click into one part of a split nucleus. The selected roi will be displayed in green. Now click into the second part of the nucleus. The two parts will be merged.
Volker Bäcker