A Modified Technic for the Preparation of Metaphase Chromosomes From Mammalian Cells Grown in Monolayers

The details of a method for the preparation of well spread metaphase chromosomes from human cells grown on cover slips in Leighton tubes is presented. Cells were inoculated and grown to 25–50% confluency. Colcemid was added to collect cells in metaphase. The monolayers were washed free of medium and treated with hypotonic solutions in the Leighton tubes in which they were grown. The coverslips were fixed and air dried and stained immediately or stored in the Leighton tube for future histochemical or immunofluorescence tests.This modification of standard chromosome harvest technics eliminates trauma to the cells caused by the manipulations previously employed. It prevents the loss of metaphase cells and cell breakage which occurs as a result of multiple centrifugations and the preparation of cell spreads on glass slides or coverslips required by standard harvest procedures. It provides information regarding mitotic indices, chromosome number and structure with relative speed, ease and less cost.