A novel method for depositing erythroid cells onto glass slides for fetal cell analysis

Background: We have developed a method for selecting erythroblasts from blo od, the first step toward identifying fetal cells in maternal blood for dia gnostic purposes. Because the selection method results in a large number of positive cells, we needed to develop new methods to deposit the cells onto slides and to modify in situ hybridization procedures to enable detection of fetal cells. Methods: We utilized Nunc flaskettes to increase the slide surface area ava ilable for cell deposition, The ability of erythroid lineage cells to adher e to several surface modifications was examined. In situ hybridization meth ods were tested to find the best approach that is compatible with these cel l preparations. Results: The best glass slide coating for erythroid cells was found to be a n antibody to glycophorin A, a red cell surface antigen. We were able to ge t excellent in situ hybridization signals in cells on flaskettes by modifyi ng fixation and pretreatment parameters. Conclusions: The methods described here appear to be the best way of attach ing a large number of erythroid lineage cells to slides and of detecting th em by in situ hybridization. (C) 2001 Wiley-Liss, Inc.