Project description:RapidAIM combines an optimized culturing model, which maintains the taxonomic and functional profiles of the human gut microbiome in vitro, and a high-throughput metaproteomics workflow to gain deep functional insights into microbiome responses. Here we exemplify the described RapidAIM 2.0 workflow with demostration study, in which we evaluated the use of different human stool sample collection and processing workflows on individual gut microbiome responses to kestose, a prebiotic oligosaccharide. Four different stool sample processing strategies, namely gauze filtration (G), 100 µm vacuum filtration (V), 100 µm spin tube filtration (S) and 100 g spinning (100g) workflows were tested. A commercial kit GutAlive® was also evaluated following the manufacturer’s manual for its performance on live microbiome sample collection and preservation under room temperature for 24 hours and 72 hours before being processed using the 100 µm vacuum filtration protocol.