*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Folks,

I need your advice on what to do with a manuscript. The story is, we have been working 
on a quantification protocol for a while with a student on her large-scale imaging project. 
We spent some brain power on it, so at the end, we have decided to publish it as a small 
methods paper. the novelty of our approach was applying the Nyquist sampling rate to 
the target object size, rather than the confocal system output AND adequate post-
processing. we have shown that 1)our suggested algorithm works well in terms of 
preserving the number of counts acquired, compared to higher sampling rates; and allows 
to  keep image size/sampling density/imaging time about several fold lower than you 
would do standard 2)if you neglect proper sampling rates (linked to the object size!) or 
skip processing, your results suck.

we have sent the paper to two journals, and received three sets of comments

reviewer 1: overall correct, but...nothing new .. AND(!!)... In many studies, 
photobleaching is a major determinant of the spatial sampling rate to use 

other journal

reviewer 2:
..a pipeline for speckle counting on the CellProfiler example page that seems relevant... 
and..the use of passive voice throughout makes it a difficult and dry read..

reviewer 3:

The authors fail to demonstrate that using this method increases the accuracy of their 
quantitation (We were aiming at preserving the accuracy and minimizing the effort!!). 

This method is not broadly applicable (???, almost  every lab has to quantify images).  

My main idea behind submitting the manuscript was, that its always nice to have an 
example of a working protocol, and sampling rate is something often neglected (see 
comments from reviewer 1). I have seen tons of very smart grad students, who need to 
do quantification, but end up performing manual counting on their images, since adapting 
existing protocols is beyond their available effort. On the other hand, I am personally not 
qualified to go deep into physics and math behind sampling according to the PSF of the 
system vs sampling based on object density. However, I know that sampling below 
Nyquist is hot in medical imaging field now.

I can not publish the full method within the main paper from the study. Quit? Try other 
microscopy journals? Publish on the Core's webpage?