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Perfusion System for Modification of Luminal Contents of Human Intestinal Organoids and Realtime Imaging Analysis of Microbial Populations
- Source :
- Micromachines, Micromachines, Vol 13, Iss 131, p 131 (2022), Micromachines; Volume 13; Issue 1; Pages: 131
- Publication Year :
- 2021
-
Abstract
- Intestinal organoids are 3D cell structures that replicate some aspects of organ function and are organized with a polarized epithelium facing a central lumen. To enable more applications, new technologies are needed to access the luminal cavity and apical cell surface of organoids. We developed a perfusion system utilizing a double-barrel glass capillary with a pressure-based pump to access and modify the luminal contents of a human intestinal organoid for extended periods of time while applying cyclic cellular strain. Cyclic injection and withdrawal of fluorescent FITC-Dextran coupled with real-time measurement of fluorescence intensity showed discrete changes of intensity correlating with perfusion cycles. The perfusion system was also used to modify the lumen of organoids injected with GFP-expressing E. coli. Due to the low concentration and fluorescence of the E. coli, a novel imaging analysis method utilizing bacteria enumeration and image flattening was developed to monitor E. coli within the organoid. Collectively, this work shows that a double-barrel perfusion system provides constant luminal access and allows regulation of luminal contents and luminal mixing.
Details
- ISSN :
- 2072666X
- Volume :
- 13
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Micromachines
- Accession number :
- edsair.doi.dedup.....0b2c752742b733ecedf4eb44998d4584