@@ -22,42 +22,42 @@ Once the site has been added, updating Fiji should cause it to download and inst
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@@ -22,42 +22,42 @@ Once the site has been added, updating Fiji should cause it to download and inst
## How it works?
## How it works?
1. Open Fiji
### Step 1. Open Fiji
2. Open *Plugins -> Eigen_PSF -> Eigen_PSF*
### Step 2. Open *Plugins -> Eigen_PSF -> Eigen_PSF*


3.**Drag and drop the images** you want to open in the left pane.
### Step 3. **Drag and drop the images** you want to open in the left pane.


4.**Display the selected images** by clicking on the eye (show) in the left pane.
### Step 4. **Display the selected images** by clicking on the eye (show) in the left pane.


5. Click on the parameters button and set the parameters. In particular, pay attention to :
### Step 5. Click on the parameters button and set the parameters. In particular, pay attention to :
-**Set 2D/3D option**.
-**Set 2D/3D option**.
-**Set the PSF width**. You can also do it by drawing a square around a PSF and by clicking on get_ROI.
-**Set the PSF width**. You can also do it by drawing a square around a PSF and by clicking on get_ROI.
-**Set the number of EigenPSFs** (this can be done afterwards as well).
-**Set the number of EigenPSFs** (this can be done afterwards as well).


6. Click on *Detect beads*. This may take some time. When finished, you can click on the image pane, and ROI should appear around each detected microbead.
### Step 6. Click on *Detect beads*. This may take some time. When finished, you can click on the image pane, and ROI should appear around each detected microbead.


7. The PSFs that are set as *valid* are displayed in green. Those in red are considered as *invalid* because, they are out of the distribution of observed beads. You can change the *valid*/*invalid* status by *ctrl + shift + click* or by clicking on the table at right. Note: the table contains the main beads properties. The lower the bead quality, the more likely it is for it to be a correct bead (i.e. no overlap with other beads, out of distribution features,...).
### Step 7. The PSFs that are set as *valid* are displayed in green. Those in red are considered as *invalid* because, they are out of the distribution of observed beads. You can change the *valid*/*invalid* status by *ctrl + shift + click* or by clicking on the table at right. Note: the table contains the main beads properties. The lower the bead quality, the more likely it is for it to be a correct bead (i.e. no overlap with other beads, out of distribution features,...).
8. Click on Preprocess Patches. This will do a fine PSF registration on the detected patches and remove the background.
### Step 8. Click on Preprocess Patches. This will do a fine PSF registration on the detected patches and remove the background.


9. Click on Compute EigenPSF. You can also show them. You should get a result like the one below. Here we made a montage with 5 eigenPSFs extracted from the valid microbeads.
### Step 9. Click on Compute EigenPSF. You can also show them. You should get a result like the one below. Here we made a montage with 5 eigenPSFs extracted from the valid microbeads.


10. Finally, you can display the processed PSFs after registration, background removal and projection on the eigenPSF basis.
### Step 10. Finally, you can display the processed PSFs after registration, background removal and projection on the eigenPSF basis.