Karyotype analysis in my research project

Good day everyone. I am new to molecular cytogenetics that I need your help in my karyotype analysis. My objective is to construct the karyotype of my species (2n=32) using 5s and 45s rDNA probes shown in red and green signals, respectively. I am struggling with finding the pair for the chromosomes by merely checking the morphology and centromere location. Literature has shown that the species I am working on has a symmetric karyotype. Is there a way by image analysis that I could pair my chromosomes? Let us say intensity of each chromosome? How shall I tackle this problem? Do chromosome pairs have almost the same intensity when viewed under the fluorescence microscope? How could I better enhance this image? Thank you very much for your help. Normal with signals 0409-7.tif (4.1 MB)