Detecting subtle subterranean movement via laser speckle imaging

A diversity of organisms live within underground environments. However, visualizing subterranean behavior is challenging because of the opacity of most substrates. We demonstrate that laser speckle imaging, a non-invasive technique resolving nanometer-scale movements, facilitates quantifying biological activity in a granular medium. We monitored fire ants (Solenopsis invicta) at different developmental stages, burial depths (1-5 cm) and moisture fractions (0 and 0.1 by volume) in a container of 0.7 mm glass particles. Although the speckle pattern from the backscattered light precludes direct imaging of animal kinematics, analysis of integrated image differences revealed that spiking during ant movement increased with the developmental phase. Greater burial depth and saturation resulted in fewer and lower magnitude spikes. We verified that spiking correlated with movement via quasi-2D experiments. This straightforward method, involving a laser and digital camera, can be applied to laboratory and potentially field situations to gain insight into subterranean organism activities.