A novel method for depositing erythroid cells onto glass slides for fetal cell analysis.
BACKGROUND
We have developed a method for selecting erythroblasts from blood, the first step toward identifying fetal cells in maternal blood for diagnostic 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 available for cell deposition. The ability of erythroid lineage cells to adhere to several surface modifications was examined. In situ hybridization methods were tested to find the best approach that is compatible with these cell preparations.


RESULTS
The best glass slide coating for erythroid cells was found to be an antibody to glycophorin A, a red cell surface antigen. We were able to get excellent in situ hybridization signals in cells on flaskettes by modifying fixation and pretreatment parameters.


CONCLUSIONS
The methods described here appear to be the best way of attaching a large number of erythroid lineage cells to slides and of detecting them by in situ hybridization.
Collarini
E J
EJ
Chief Technology Office, Roche Diagnostics, Berkeley, California, USA.
Nagy
D
D
Cain
C A
CA
Gammon
D
D
Schueler
P A
PA
Mahoney
W C
WC
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects; D002499:Centrifugation, Density Gradient; D004900:Erythroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000276:immunology; D005331:Fetomaternal Transfusion; D005865:Gestational Age; D005914:Globins / Q000235:genetics / Q000378:metabolism; D006021:Glycophorins / Q000276:immunology; D006801:Humans; D019044:Image Cytometry / Q000379:methods*; D017403:In Situ Hybridization / Q000379:methods; D066298:In Vitro Techniques; D008297:Male; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D013048:Specimen Handling / Q000379:methods; D016707:Tissue Fixation
D000911:Antibodies, Monoclonal; D006021:Glycophorins; D005914:Globins