Background Phakic intraocular lens (pIOL) implantation has been a popular means for the treatment of high ametropia. Measurements of ciliary sulcus diameter is important for pIOL size determining. But till now, no perfect system can directly measure it. The present study was to evaluate the accuracy, repeatability and reproducibility of direct sulcus diameter measurements obtained by a full-scale 50-megahertz (MHz) ultrasound biomicroscopy (UBM). Methods A fresh cadaver human eye with a scale marker inserted through the posterior chamber plane from 3 o'clock to 9 o'clock meridian and 30 randomly selected eyes from 30 normal subjects were scanned by full-scale 50-MHz UBM in horizontal meridional scan plane. The distance between the scales and the whole length of the marker inside the cadaver eye were measured by the same observer using the "built-in" measurement tools and the indicating error of instrument was calculated. Reproducibility of the measurement was evaluated in 30 eyes by 2 operators using Blander and Altman plot test. Repeatability was evaluated from 10 successive eyes randomly selected from the 30 eyes by one operator. Results On a scale of 1 mm, the greatest indicating error was 40 mu m; the mean largest indicating error of 1 mm scale from the 10 images was (26 +/- 14) mu m; on a scale of 11 mm, the greatest indicating error was 70 mu m; the error rate was 0.64%. The mean length of the needle inside the eye of the 10 images was 11.05 mm, with the mean indicating error of 47 mu m, the average error rate was 0.43%. For ciliary sulcus diameter measurements in vivo, the coefficient of variation was 0.38%; the coefficients of repeatability for intra-observer and inter-observer measurements were 1.99% and 2.55%, respectively. The limits of agreement for intra-observer and inter-observer measurement were -0.41 mm to 0.48 mm and -0.59 mm to 0.58 mm, respectively. Conclusion The full-scale 50-MHz UBM can be a high accuracy and good repeatability means for direct measuring the ciliary sulcus diameter and useful for size determining of posterior chamber pIOL.