Preparing Retinal Organoid Samples for Transmission Electron Microscopy.
J Vis Exp
; (208)2024 Jun 07.
Article
en En
| MEDLINE
| ID: mdl-38912821
ABSTRACT
Retinal organoids (ROs) are a three-dimensional culture system mimicking human retinal features that have differentiated from induced pluripotent stem cells (iPSCs) under specific conditions. Synapse development and maturation in ROs have been studied immunocytochemically and functionally. However, the direct evidence of the synaptic contact ultrastructure is limited, containing both special ribbon synapses and conventional chemical synapses. Transmission electron microscopy (TEM) is characterized by high resolution and a respectable history elucidating retinal development and synapse maturation in humans and various species. It is a powerful tool to explore synaptic structure in ROs and is widely used in the research field of ROs. Therefore, to better explore the structure of RO synaptic contacts at the nanoscale and obtain high-quality microscopic evidence, we developed a simple and repeatable method of RO TEM sample preparation. This paper describes the protocol, reagents used, and detailed steps, including RO fixation preparation, post fixation, embedding, and visualization.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Retina
/
Organoides
/
Microscopía Electrónica de Transmisión
Límite:
Animals
/
Humans
Idioma:
En
Revista:
J Vis Exp
Año:
2024
Tipo del documento:
Article
Pais de publicación:
Estados Unidos