Effect of sample treatment on the elastic modulus of locust cuticle obtained by nanoindentation

Cuticle is one of the most abundant, but least studied, biological composites. As a result, it has contributed very little to the field of biomimetics. An important step to overcome this problem is to study cuticle biomechanics by means of accurate mechanical measurements. However, due to many reasons, mechanical testing on fresh cuticle specimens is not always possible. Hence, researchers often use stored specimens to measure properties of arthropod cuticle. Our knowledge about the influence of different treatment methods on cuticle properties is currently very limited. In this study, we investigated the effect of freezing, desiccation, and rehydration on the elastic modulus of the hind tibial cuticle of locusts obtained by nanoindentation. We found that all the mentioned treatments significantly influence cuticle properties. This is in contrast to previous reports suggesting that freezing did not significantly influence the elastic modulus of native cuticle specimens tested in bending. In the light of our data, we suggest that changes of the elastic modulus of cuticle are not solely due to changes of the water content. Our results provide a platform for more accurate measurements of cuticle properties.