Two-dimensional (2D) echocardiography is the cornerstone of noninvasive evaluation of the cardiac patient, and often involves estimating left atrial (LA) size. However, 2D echocardiographic methods of estimating LA size have been inadequately described, and most reference intervals are based on M-mode echocardiographic measurements. We determined reference intervals for 4 different 2D echocardiographic methods of estimating LA size in adult (> or =9-month-old) dogs without cardiovascular disease. Thirty-six dogs, placed in right lateral recumbency, were examined by 2D echocardiography. The left atrium was measured at specific time points in the cardiac cycle. Measurement methods were LA diameter in short axis, LA diameter in long axis, LA circumference in short axis, and LA cross-sectional area in short axis. Comparisons of these LA dimensions to appropriate aortic dimensions provided body weight-independent estimates of LA size. We found strong associations of LA dimensions with body weight (r2 = .76-.88). Comparable body weight-independent 2D echocardiographic estimates of LA size in short axis exceeded historical M-mode reference intervals. These data provide echocardiographers with reference intervals for 2D echocardiographic estimates of LA size in adult dogs.