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Brain Organoids

Brain Organoids

Brain organoids are 3D tissue models representing one or more regions of the brain. They can overcome the shortcomings of conventional post-mortem and animal brain models to produce clinically relevant results.

Brain organoids are derived from human-induced pluripotent stem cells (hiPSCs). When cultured with the appropriate media containing the combination of signaling factors, the stem cells differentiate into various neural cells that mature over time to resemble structures of brain regions such as the forebrain (cerebral) or mid-brain.

Cerebral organoids have great potential for understanding brain development and neuronal diseases. They can also be used for investigating genetic disorders and the effects of compounds. Nevertheless, capturing the uniqueness of the human brain requires functional assays and high-content imaging systems.

Methods for monitoring and developing cerebral organoids

Here we describe the methods for monitoring iPSC-derived 3D cerebral organoids and testing their functional activity by recording and analyzing calcium oscillations. Brightfield imaging with artificial intelligence (AI)-based segmentation can help monitor the quality of developing organoids by tracking the growth in diameter and shape. The neuronal activity of brain organoids can be determined from calcium activity. Confocal imaging reveals calcium activity to determine the maturity of the neurons. Furthermore, the cellular organization can be monitored via confocal imaging with differential staining.

Featured scientific poster: Monitoring organoid development and characterization of calcium oscillation activities in iPSC-derived 3D cerebral organoid

Cerebral organoids were developed from induced pluripotent stem cells (iPSC) using established methods. Over a period of 4-12 weeks of development, we monitored the size and morphology of the developing brain microtissues using our AI-based analysis tools, IN Carta® Image Analysis Software, for defining the size and shape of the tissues. Selected microtissues were analyzed by confocal imaging during different phases of development for cell organization and expression of neuronal markers. For detection of functional activities, organoids were loaded with calcium-sensitive dye, and then Ca2+ oscillations were recorded with the ImageXpress® Confocal High-Content Imaging System. We show that high-content imaging paired with AI-based analysis used with 3D cerebral organoids is a promising tool for compound screens and toxicity evaluations.


Calcium imaging of whole brain organoids

Calcium imaging of whole brain organoids – Starting from week 4, we observed calcium activity in cerebral organoid loaded with FLIPR Calcium 6 dye. (top-right). Frequency of calcium activity was low, suggesting that the neurons in the organoid at that time were still immature. By week 13, the calcium activity appeared more synchronous, suggesting that neurons are interconnected in a functional network (bottom-right).


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More great resources

Learn how high-content imaging and analysis tools can improve the accuracy of your cerebral organoid studies:



Resources for Brain Organoids

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Application and Analysis of Organoid Systems

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