OrganoidChip allows for rapid and blur-free imaging by facilitating hydrogel-free immobilization.
We used confocal imaging to demonstrate the ability of the OrganoidChip to enable higher-resolution images. Images show better optical segmentation, and the ability of resolving single cells in an organoid. 4 d). Co-localized EthD-1 and Hoechst stained nuclei can be resolved, which could increase the accuracy of viability measurement. The future implementation of 3D segmentation using AI algorithms can provide more accurate estimates of cellular viability on larger screens.
We then measured the effect DOX treatment had on the beating kinetics in cardiac organoids. We used calcium fluorescence to do this as it was shown to be an accurate approximation for the action potentials of cardiomyocytes32. Calcium imaging was beneficial in determining beating and contraction parameters, as brightfield images cannot detect smaller beating portions. This is especially true when organoids are compromised by drug treatment.
We observed some variability in the spontaneous contraction behaviour and beating kinetics of cardiac organoids when assessing drug effects. This variability can skew any averaged parameter values across organoids, and not reflect the effects of treatment conditions on organoid’s health. In order to overcome this problem, we measured the beating of each organoid both on and off chip. Drug-induced functional results are reported as the averages of fractional differences of each organoid’s beating kinetics parameter, measured 48 h after treatment, both on the chamber slide, and on the chip. 3).
Source:
https://www.nature.com/articles/s41598-023-38212-8
