Acoustic sensor for monitoring adhesion of Neuro-2A cells in real-time

Neuronal adhesion plays a fundamental role in growth, migration, regeneration and plasticity of neurons. However, current methods for studying neuronal adhesion cannot monitor this phenomenon quantitatively in real-time. In this work, we demonstrate the use of an acoustic sensor to measure adhesion of neuro-blastoma cells (Neuro-2A) in real-time. An acoustic sensor consisting of a quartz crystal sandwiched between gold electrodes was placed in a flow cell and filled with 600 microl of phosphate buffered saline (PBS). Two sets of in vitro experiments were performed using sensors that had uncoated gold electrodes and sensors that were coated with a known neuronal adhesion promoter (poly-l-lysine or PLL). The instantaneous resonant frequency and the equivalent motional resistance of the acoustic sensor were monitored every second. Cell Tracker was used to confirm neuronal adhesion to the surface. Addition of 10 microl of media and Neuro-2A cells into the above set-up elicited exponential changes in the resonant frequency and motional resistance of the quartz crystal with time to reach steady state in the range of 2-11 h. The steady-state change in resonant frequency in response to addition of neurons was linearly related to the number of Neuro-2A cells added (R2=0.94). Acoustic sensors coated with the adhesion promoter, PLL showed a much higher change in resonant frequency for approximately the same number of neurons. We conclude that the acoustic sensor has sufficient sensitivity to monitor neuronal adhesion in real-time. This has potential applications in the study of mechanisms of neuron-substrate interactions and the effect of molecular modulators in the extra cellular matrix.