Label-free imaging tracks lipid storage in living liver organoids

by SPIE

edited by Sadie Harley, reviewed by Andrew Zinin

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Fat accumulation in liver cells is central to several metabolic disorders, but observing the process in living tissue models remains difficult. Fluorescent labels and stains can provide detailed images, but they may disrupt cells or capture only selected time points. A label-free approach for tracking changes in lipid storage over time in three-dimensional mouse liver organoids is reported in Biophotonics Discovery.

The approach was developed by a research team led by YongKeun Park at the Korea Advanced Institute of Science and Technology. The team used holotomography, an imaging technique that maps differences in how light travels through a sample. Because lipid droplets have distinct optical properties, the method can identify and measure them without fluorescent dyes. Repeated 3D imaging adds a time dimension, allowing the same organoid to be monitored as lipid droplets form and change.

Imaging deep inside organoids presents another challenge: image quality and optical properties vary with depth. To address this, the team developed a depth-adaptive segmentation method that adjusts how lipid droplets are identified in different regions of the organoid. This allowed them to measure droplet number, volume and shape and to estimate lipid concentration and dry mass from refractive-index measurements.

The researchers then exposed hepatic organoids to three fatty acids: oleic acid, linoleic acid and palmitic acid. Oleic and linoleic acids both increased lipid accumulation while preserving the overall structure of the organoids, but they produced different patterns.

Oleic acid primarily increased the size of lipid droplets, producing fewer, larger droplets with a broader range of sizes. Linoleic acid instead increased the number of droplets, resulting in a more uniform population of smaller droplets. Palmitic acid produced a markedly different response, causing rapid structural deterioration of the organoids.

The findings show that measuring total lipid content alone can obscure important differences in how cells respond to fatty acids. By quantifying droplets at successive time points in living organoids, the approach distinguishes accumulation patterns dominated by increasing droplet number versus increasing droplet size.

More broadly, the study demonstrates how label-free 4D imaging can turn organoid experiments from a series of endpoint measurements into longitudinal observations of cellular change. In this study, 3D images were acquired at six-hour intervals. That capability could help researchers investigate lipid metabolism and other dynamic processes in living tissue.

More information

Jimin Cho et al., Label-free 4D holotomography with depth-adaptive segmentation for quantitative analysis of lipid droplet dynamics in hepatic organoids, Biophotonics Discovery (2026). DOI: 10.1117/1.bios.3.3.035005

Key medical concepts

Lipid DropletsLinoleic AcidOleic AcidPalmitic Acid

Clinical categories

Gastroenterology Provided by SPIE Who's behind this story?

Sadie Harley

BSc Life Sciences & Ecology. Microbiology lab background with pharmaceutical news experience in oil, gas, and renewable industries. Full profile →

Andrew Zinin

Master's in physics with research experience. Long-time science news enthusiast. Plays key role in Science X's editorial success. Full profile →

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