Hello dear Mario. 

I use ImageJ to count particles, it is a program with open access and easy to use. There is a link to download the program.

Best regards

https://imagej.nih.gov/ij/

Images from tangential view would be more informative if taken against a dark screen. An alternative is removing spines by a fine trimming shear and take the record relative to area of clatode and conditioning. structure and bending relative to a force can then be recorded with density.

Can't give you the name of a software to count, but some hints about preprocessing:

* a sheet of black paper would have been able to mask the background.

* And you may 'fiddle' a bit with the angle of illumination to get the spines even

   brighter.

Hope this helps

Hi Mario,

Thanks for posting the image! A very challenging problem indeed. I definitely agree with U. Dreher's comments on illumination and imaging setup care. Fortunately, with the current image, I was able to use our new software MIPAR (available at http://MIPAR.us) to quickly build a recipe which quite nicely found and counted your spines. There are a few errors, but you can easily clean those up manually if needed (even if you have multiple images) using MIPAR's manual editing tools. I have attached:

Please let me know of any questions. In total I counted 330 spines.

Cheers,

John

Hello to all

Thank you very much for your guidance. Let me try all of this options and I'll answer what works best for my investigation.

I think that all of them are useful for the purpose. I'll accept another ideas, if there are.

Regards.

Mario Sandoval