Semi-automated image acquisition and automatic image quantification methods for liver Organ-Chips.

Toxicant exposure can induce acute or chronic alterations in cellular numbers, morphology, and cell function. The quantification of these parameters can provide valuable information regarding a toxicant's effect and/or mechanism of action in organ-on-a-chip toxicity testing platforms. Unfortunately, manual quantification can be variable and time consuming. Additionally, the unique designs of Organ-Chips make automated imaging difficult as current microscopes were not specifically designed for Organ-Chip use. The development of semi-automated and automated imaging and quantification procedures greatly increases the quantity and quality of collected data. Using Emulate's transparent liver Organ-Chip (Liver-Chip) in combination with Keyence's bench-top BZ-X700 All-in-one fluorescence microscope we have developed semi-automated imaging and automated quantification methods for nuclei, mitochondrial viability, and apoptosis. The methods described herein provide alternative imaging options to more costly and space consuming microscopes while still providing necessary features for Organ-Chip evaluation. We were able to detect significant decreases in nuclear number and mitochondrial membrane potential, and significant increases in apoptosis with a model hepatotoxic compound, benzbromarone. These methods have greatly reduced the time and increased the quality of cell number/function data acquisition and demonstrated that these automated quantification methods can detect changes resulting from chemical exposure.