On Sun, 15 Jan 1995, k.a. rogers wrote:
 
> I have recently run into some rather obnoxious comments from a reviewer
> of one of my manuscripts, who implied that we had been less than honest
> in our presentation of our images.  So I put the question to you all.
> How much enhancement is too much?  What is acceptable?
>
> In this particular case, we had a double labelled FISH sample, using
> propidium iodide and FITC.  The image was enhanced by setting all
> green pixels below ca. 40 to 0 (to remove backgound FITC) and then enhancing
> contrast by mapping the remaining pixel values over the entire 256 range for
> each channel.  No other manuipulation was carried out.
>
> If we are out of line with this approach, I would appreciate some
> guidance.  If the approach is appropriate, I would like to see your
> comments. I hope to get back to the editor by weeks end.
>
 
 
Sounds pretty reasonable to me.  My opinion is that if you detail your
manipulations in the manuscript, you are pretty much covered.
Thresholding and contrast stretching are standard enhancement methods; if
the reviewer has a specific criticism in their application in your
specific result because of some particular situation, then you can address
them specifically.  If the comment is a general disparagement of image
enhancement, then I would suggest you include references to instances
where this technique was applied without problems in similar
circumstances.
 
A general disparagement of image processing in confocal microscopy is a
bit unsettling, since all 3D visualizations are rather heavily processed...
 
 
 
billo
 
(William R Oliver, MD
 Digital Image Processing Laboratory
 Division of Quantitative Pathology
 Department of Cellular Pathology
 Armed Forces Institute of Pathology
)