Caenorhabditis elegans revisited by atomic force microscopy - Ultra-structural changes of the cuticle, but not in the intestine after treatment with Combretum mucronatum extract

Abstract

Assessing the internal morphology of Caenorhabditis elegans by a topographical technique like atomic force microscopy (AFM) is a challenging process. As a prerequisite for a successful image acquisition, direct contact between the structure of interest and the AFM probe needs to be established. To gain this insight into the morphology of cuticle and intestine in C. elegans before and after treatment with a tannin-enriched hydro-ethanolic extract from Combretum mucronatum, we developed an approach based on polyethylene glycol embedding, ultra-sectioning, de-embedding and hexamethyldisilazane-dehydration prior to measuring in ambient conditions by intermittent contact mode AFM. The used experimental protocol allowed a facile and fast insight into the ultrastructure of treated versus untreated C. elegans individuals, directly leading to the identification of treatment-associated morphological alterations in the cuticle but not the intestine of C. elegans. Additionally, the presented ultra-microtomy based protocol could allow future insight into virtually any tissue or organism by AFM.

Keywords: Anthelmintic; Atomic force microscopy; Caenorhabditis elegans; Polyethylene glycol embedding; Tannins; Ultra microtomy.