To the flow list, A colleague of mine asked kindly if I could post this question on her behalf. Dear All, We are trying to determine DNA content of cells (sperm) from an animal that only produces a few hundred at at time. At this stage we are using the Propidium Iodide method using chick erythrocyte nuclei (CEN) as the internal standard. The difficulty is that we can not determine the nuclei number with a haemaocytometer and have just tried to reduce the number of CEN. As a result from run to run we do not have exactly the same number of nuclei and the data differ. I expect this is because of the relationship between PI and nuclei with staining differing among trials. When we did this experiment many years ago with a UV source and DAPI as the stain we did not seem to have this problem. The reason that we went to fluorescence is that we no longer had a UV flow cytotmeter. So my question is- should we give up trying to use PI and go back DAPI in the hopes that we can accomplish what we need to do and if we can find a UV machine. Any advice is greatly appreciated. Best wishes Maria Byrne Cheers, Tara McDonald ******************************************************** Tara McDonald (BSc. Hons) Flow Cytometry Facility Centenary Institute of Cancer Medicine and Cell Biology Locked Bag No. 6 NEWTOWN NSW 2042 AUSTRALIA Tel: 612 9565 6140 Fax: 612 9565 6105 ********************************************************
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:08 EST