However, these assays typically cannot capture the interactions between the host and the bacteria on a microscopic scale. In order to discover the virulence determinants, different mutants of a pathogen of interest as well as the worms are generated and the interactions between the bacteria and the host (worm) are studied through survival curves and bacterial colonization assays. 1–7 In particular, this roundworm, owing to its genetic tractability and evolutionarily conserved innate immunity is an established model organism for the investigation of virulence mechanisms of pathogens. elegans is a bacterivorous organism with considerable genetic similarity to humans and is used as a model for studying the relation between microbiota and aging and health/diseases. We show the advantage of single-worm resolution automated analysis for bacterial load studies and anticipate that the methods described in our work can be easily implemented in existing microfluidic solutions to enable thorough studies of bacterial proliferation. We present an automated bivariate histogram analysis of the volumes and intensities of the bacterial spots for each worm and find that, as the worms age, the bacterial load in their hindguts increases. High-resolution z-stack fluorescence images of the gut bacteria in adult worms, which were loaded in the microfluidic chip and subsequently fixed, were analyzed using IMARIS software and 3D reconstructions of the intestinal bacterial load in the worms were obtained. ![]() Use of a microfluidic chip based on a thin glass coverslip substrate allows investigation of the intestinal bacterial load using a Spinning Disk Confocal Microscope (SDCM) equipped with a high-resolution objective (60×). ![]() elegans) of a fluorescent strain of the worm's bacterial food ( Escherichia coli ( E. We study the presence within the worm Caenorhabditis elegans ( C.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |