the colours that paint almost all my chromosomes are violet blue and green. Is there any problem with the procedure or something wronge with the programm-microscope settings? Moreover when i want to make karyotyping (i use cytovision softwarware), i open the fluomap and scan the chromosomes i find piecies of differrent chromosomes (in normal sample) why is happend that?
Hi,
from these information it is hard to say. From these information, there are some possibilities. Maybe if you know, that your microscope can separate different spectra (check, that you are you using three color filter with different emission spectrum), maybe check quality of your probe or concentration for NICK translation. If some pieces of chromosomes occured (I suppose you see small pieces, that you dont want), try additional washing steps (e.g.-2xSSC or you can use ssDNA for decreasing backround and cutted pieces of chromosome). Incerasing of temperature for denaturation is helping too, but be careful for ovedenaturating.
Dear Katerine,
remembering your question, I found one nice review. Maybe it is familiar to you, anyway good luck.
Vaclav
http://www.hh.um.es/pdf/Vol_19/19_1/Liehr-19-229-237-2004.pdf
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