Micro-moulded substrates for the analysis of structure-dependent behaviour of nematodes

Abstract

AbstractMicrostructures, consisting of networks of channels of rectangular cross sections (50 μm high, 40-200 μm wide), were formed in 5 mm square areas on transparent substrates made of silicon rubber. An experimental set-up using the network sealed with a flat glass plate has the potential to function in a way similar to the pore space in soil, and is therefore useful for studies of nematode migration. The set-up allowed the migratory activity of nematodes in water-filled, porous and transparent microstructures to be observed with a microscope. By means of substrates with two different channel dimensions, the structure-dependent behaviour of second-stage juveniles of Meloidogyne incognita was visually demonstrated. Their behaviour was examined on the basis of the migration patterns obtained by superimposing recorded serial images of individual juveniles. In a micro-channel network with 40 μm high channels of 200 μm wide elements, juveniles showed marked activity in migration, forming consistent zigzag patterns spread over the network area. In contrast, in a micro-channel network with 80 μm high channels of 400 μm wide elements, migration showed thick, sparse patterns, restricted around the area where the juveniles were initially deposited. This comparison showed that M. incognita juveniles in a narrow, fine network tended to migrate actively and, in contrast, those in a wide, coarse network were prevented from migrating by the network configuration.