A micromethod for chromosome preparation from individual hematopoietic colonies cultured in methylcellulose.
We report a micromethod for chromosome preparation from individual hematopoietic colonies cultured in methylcellulose. The entire process was carried out on poly-Lysine (PL)-coated slides. Individual colonies were transferred into 10 microliter of 0.075 M KCl and placed on PL-coated slides. After hypotonic treatment of the colony cells and their attachment to the slides, the cells were fixed by a three-step procedure as follows: addition of a 30% fixative (3:1 methanol:acetic acid) diluted with the hypotonic solution, addition of 20% ethanol, and subsequent immersion of the slides in a 100% fixative. The slides were flame dried and Giemsa stained. Q- and G-banding techniques also were used. These procedures provided analyzable chromosome preparations, even from colonies containing fewer than 50 cells.
D000368:Aged; D001853:Bone Marrow; D002469:Cell Separation; D002478:Cells, Cultured; D002871:Chromosome Banding; D002875:Chromosomes; D003584:Cytological Techniques; D005260:Female; D006098:Granulocytes; D006412:Hematopoietic Stem Cells / Q000648:ultrastructure*; D006801:Humans; D066298:In Vitro Techniques; D015470:Leukemia, Myeloid, Acute / Q000235:genetics; D008297:Male; D008747:Methylcellulose
D008747:Methylcellulose