Operational Modes and Speed Considerations of an Acoustic Droplet Dispenser for Mass Spectrometry.

Acoustic ejection mass spectrometry is a recently developed concept in which low nanoliter-volume sample droplets are acoustically dispensed from microtiter plate wells into a continuous fluid transfer open-port interface for subsequent ionization at atmospheric pressure. This manuscript focuses on the acoustic droplet dispensing component of a prototype system, in particular the well-to-well sampling speed, droplet volume calibration, precision, reproducibility, and various modes of operation this device enables. A new method to measure the volume of individually dispensed droplets is presented to both aid method validation and potentially assist in the tuning of acoustic dispense parameters for samples having a wide range of viscosities and surface tensions. Acoustic dispensing modes of operation discussed are high-speed, well-to-well dispensing of individual nanoliter-scale droplets from microtiter plates, including the first demonstration of 1536-well compatibility using this approach. Multiple nanoliter-volume droplets per sampling event to increase detection dynamic range is described, and a "continuous infusion" mode to provide a low sample consumption sustained steady-state signal for analyte detection optimization, improved ion statistics and signal-to-noise ratio (S/N), or time for in-depth tandem mass spectrometry of the components in a sample is presented. The concept of "bar coding" using combinations of dispensed droplet patterns to register well-plate position to specific mass spectral signals is introduced, as well as judicious well-plate sample layout to enable assay "multiplexing" as a means to maximize well-to-well sample analysis throughput, is also demonstrated.