Gastrointestinal (GI) diseases, such as inflammatory bowel disease, Hirschsprung's disease, and chronic intestinal inflammation, could have an impact on people worldwide. These diseases
could affect the entire GI tract or only specific parts and primarily induce disruptions in intestinal motility. Zebrafish is an excellent animal model for studying intestinal motility due to its transparency
and genetic similarity to humans. Spatiotemporal maps (STMap) is a technique used to analyze intestinal motility by quantifying the movement captured in time-lapse images. However, previous
STMap research has focused on specific intestinal regions. In this study, a subtraction-background technique was employed to map intestinal motility by identifying the intestinal structure in zebrafish.
Additionally, a zebrafish embedding tool was developed to immobilize larvae for long-term analysis of intestinal motility. As a result, STMap analysis enhanced the quantitative assessment of motility
patterns (frequency, velocity, and amplitude) in the entire intestine of larvae. This research enhances our understanding of intestinal motility analysis resulting from GI diseases or external stimuli and
enables the evaluation of GI motility parameters including frequency, velocity, and amplitude in zebrafish larvae.