Preparing Retinal Organoid Samples for Transmission Electron Microscopy

Area of Science:

• * Stem Cell Biology
• * Neuroscience
• * Ophthalmology

Background:

• * Retinal organoids (ROs) derived from induced pluripotent stem cells (iPSCs) model human retinal development.
• * Synapse development in ROs is known, but ultrastructural details remain limited.
• * Transmission electron microscopy (TEM) is crucial for high-resolution ultrastructural analysis.

Purpose of the Study:

• * To develop a simple, repeatable protocol for preparing RO samples for TEM.
• * To enable nanoscale visualization of synaptic contacts in ROs.
• * To provide high-quality microscopic evidence of RO synaptic ultrastructure.

Main Methods:

• * Detailed protocol for RO TEM sample preparation.
• * Steps include fixation, post-fixation, embedding, and visualization.
• * Focus on achieving high-resolution imaging of synaptic structures.

Main Results:

• * A simple and repeatable RO TEM sample preparation protocol was established.
• * The method facilitates high-quality microscopic evidence of RO synaptic contacts.
• * Enables nanoscale exploration of synaptic structures in ROs.

Conclusions:

• * The developed protocol is effective for RO TEM sample preparation.
• * This method enhances the study of synaptic ultrastructure in retinal organoids.
• * Facilitates deeper understanding of retinal development and synapse maturation in vitro.