Nylon mesh chip promotes three-dimensional visualization of intestinal organoids.

Abstract

Organoids are pivotal for bridging cellular-level and organism-level biological studies; however, significant challenges persist in their three-dimensional (3D) visualization. This study presents a nylon mesh chip designed to overcome these obstacles specifically for intestinal organoids (IOs). The chip, meticulously fabricated and assembled, comprises an upper glass layer, a nylon mesh, and a lower glass layer. We cultured IOs from mouse intestinal crypts and performed fluorescent labeling on the chip. For enhanced visualization, fluorescent labeling combined with 3D reconstruction techniques was employed. Results demonstrate that the chip's structure stabilizes IOs in liquid environments. While conventional fluorescence imaging is limited by mesh interference, laser confocal 3D reconstruction achieves high-quality visualization by effectively filtering out redundant signals. The nylon mesh chip is a robust tool for 3D visualization of IOs and holds potential for other budding organoid types. This innovation is poised to advance organoid 3D visualization research.