The C-band and G-band patterns of Microtus agrestis metaphase chromosomes are described. The C-band pattern reveals constitutive heterochromatin as uniformly intenselystained areas but cannot aid in identifying autosomal pairs. The G-bands revealed by a heat renaturation method (ASG) were compared with those revealed by treatment with three proteolytic enzymes. All procedures yield apparently the same specific pattern of crossbands on the autosomes, and each autosomal pair can therefore be identified by its characteristic pattern. The constitutive heterochromatin of the sex chromosomes stains uniformly with the heat renaturation method but is subdivided into regions with different staining intensities with each enzyme treatment. A G-band karyotype for Microtus agrestis metaphase chromosomes is presented.