Dear all We are trying to sort early gastrula cells (stage 10) from xenopus laevis embryos. We know that these cells are 70-100mcm in diameter and very fragile especially if they come to an air/water interface (they grow happily in liquid). We are using a BD FACSVantageSE with a 200mcm nozzle, a sheath pressure of 3psi and a sample differential of less than 1psi. The problem is that the "cells" appear to have similar scatter properties to debris and when we sort onto a slide we can only see debris and cell fragments. We suspect that the shear pressure at the nozzle is causing the cells to break up. We would be most grateful for comments from anyone who has successfully sorted this type of cell or any general comments for protecting fragile cells from the sorter. Thanks in advance Ian Ian Titley PhD Leukaemia Research Fund Centre at the Institute of Cancer Research 237 Fulham Road LONDON SW3 6JB UK Tel Direct: +44 (0)20 7970 6048 or Tel: +44 (0)20 7352 8133 ext5134 Fax: +44 (0)20 7352 3299 E-mail: iant@icr.ac.uk
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:07 EST