Select the GFP Channel Corresponding to the Brightfield Best Focus Slice¶
Using the Stack Reduction Tool users can select the best focus brightfield channel and corresponding slices of other color channels. In this example, we select the best focus slice of the brightfield channel and corresponding slice for the green GFP channel. Selecting the best focus slice using the laplacian projection from each channel separately can result in slices representing different planes of the sample.
Step |
Function |
|---|---|
1 |
Open an acquired dataset in the MCAM™ Viewersoftware. |
2 |
Navigate to Tools > Stack Reduction. A panel will open on the right. |
3 |
On the brightfield channel, select the Laplacian focus selection method. |
4 |
Select the “Target for Cropping” option checkbox for the brightfield channel. This means that all channels will align around the best-in-focus plane identified in the brightfield channel. |
5 |
As the “Target for Cropping” was set for the brightfield channel, the GFP channel will align to the same Z-plane that was selected as the best focus from the brightfield. |
6 |
The results are saved in a folder named “projection_results_timestamp” in the provided dataset’s folder with the metadata and the Tiff files containing the processed images. |