The accurate delivery of small volumes is a critical factor in the crystallization of macromolecules as it influences the reproducibility of the screening experiments. Crystallographic screening technologies have made it possible to perform experiments using volumes as low as 50 nl. The accuracy of the dispenser has usually been calibrated by weight measurements. In this work, a simple and inexpensive fluorescence-based calibration method that is sensitive and that can be used to monitor the precision and accuracy of any liquid-handling nano-dispenser device is presented. The results suggest that the protocol described here can be useful to determine volumes ranging from 50 to 300 nl with precision. Therefore, the pipetting of volumes as low as 50 nl can be calibrated periodically to ensure that precision and accuracy are maintained. The suggested calibration protocol can be executed in 6 h per instrument, including the calibration curve, which is the most time-consuming step; the rest can be completed in approximately 2 h.
Keywords: calibration; liquid-handling nano-dispenser devices.