Hi,
I'm working with a fungal specie wich have plurinuclear cells (conidia or hyphae or protoplasts have between 1 (< 5%) to 8 nuclei however majority have 4-6 nuclei ). I have made deletion mutants and my aim is to separate only cells with one nuclei (or enriching my cell population with mononuclear cells) to obtain rapidly a genetically pure strain. Monocuclear cells appear slightlys smaller than plurinuclear cells. I have tried to separate them with filters to retain plurinuclear cells but I always find a majority of plurinuclear cells after the filter.
I have readen things about density separation. Someone did already try a method like this? Did you know other separation/selection methods?
Tkank you,
Matthieu
PS: We have a pure GFP-histone strain to test differents propositions in the lab.
you should be able to separate nicely on the basis of nuclear content using FACS, if there is a sorting facility where you work. This the fasted, most accurate way to get distinct populations of high quantity. up to 100,000 events per second.
Here is a good reference:
- Badges
- Science topic
- Similar topics
- Control Systems Engineering
- Filtering