I am using a Live\ dead staining protocol. (Link to Manuals & protocols: https://www.thermofisher.com/order/catalog/product/L7007)
Briefly, This assay stain Live bacteria in Green as well as dead bacteria in Red.
I am interested in finding the actual number of all red bacteria (dead) and Live (green) bacteria.
Please see the attached pictures as my representative pictures for analysis.
I am using a fluorescence microscopy (Nikon - Dual Band Excitation Filter Sets) in order to get this RGB pictures.
I really tried everything. but I couldn’t find or design a protocol that will count all
My starting point is to press “Color – > split channels” ( this way I can discard the blue channel that I am not interested in).
Moreover, please note that bacteria size in about 2 microns ( which is equal to 13 pixels in this picture) and their shape can vary from ellipse to rings.
one last thing - bacteria can generate 3D structures and assemble together (one bacteria above other bac
I mostly interested in counting those individual bacteria, but if there will be a protocol that I can count those 3D structures of bacteria, in addition, it will be the best.
I welcome all suggestions from the forum members,
Please help me