Investigating the motility of Dictyostelium discodeum using high frequency ultrasound as a method of manipulation

Cell motility is an essential process in the development of all organisms. The earliest stages of embryonic development involve massive reconfigurations of groups of cells to form the early body structures. Embryos are very complex systems, and therefore to investigate the molecular and cellular basis of development a simpler genetically tractable model system is used. The social amoeba Dictyostelium Discoideum is known to chemotax up a chemical gradient. From previous work, it is clear that cells generate forces in the nN range. This is above the limit of optical tweezers and therefore we are investigating the use of acoustic tweezers instead. In this paper, we present recent progress of the investigation in to the use of acoustic tweezers for the characterisation of cell motility and forces. We will describe the design, modelling and fabrication of several devices. All devices use high frequency (>15MHz) ultrasound to exert a force on the cells to position and/or stall them. Also, each device is designed to be suitable for the life-sciences laboratory where form-factor and sterility is concerned. A transducer (LiNo) operating at 24 MHz excites resonant acoustic modes in a rectangular glass capillary (100um by 2mm). This device is used to alter the directionality of the motile cells inside the fluid filled capillary. A quarter-ring PZT26 transducer operating at 20.5MHz is shown to be useful for manipulating cells using axial acoustic radiation forces. This device is used to exert a force on cells and shown to pull them away from a coverslip. The presented devices show promise for the manipulation of cells in suspension. Currently the forces produced are below that required for adherent cells; the reasons for this are discussed. We also report on other issues that arise when using acoustic waves for manipulating biological samples such as streaming and heating.