Immuno-scanning electron microscopy of islet primary cilia.
J Cell Sci
; 137(20)2024 Oct 15.
Article
en En
| MEDLINE
| ID: mdl-38804679
ABSTRACT
The definitive demonstration of protein localization on primary cilia has been a challenge for cilia biologists. Primary cilia are solitary thread-like projections that have a specialized protein composition, but as the ciliary structure overlays the cell membrane and other cell parts, the identity of ciliary proteins are difficult to ascertain by conventional imaging approaches like immunofluorescence microscopy. Surface scanning electron microscopy combined with immunolabeling (immuno-SEM) bypasses some of these indeterminacies by unambiguously showing protein expression in the context of the three-dimensional ultrastructure of the cilium. Here, we apply immuno-SEM to specifically identify proteins on the primary cilia of mouse and human pancreatic islets, including post-translationally modified tubulin, intraflagellar transport (IFT)88, the small GTPase Arl13b, as well as subunits of axonemal dynein. Key parameters in sample preparation, immunolabeling and imaging acquisition are discussed to facilitate similar studies by others in the cilia research community.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Islotes Pancreáticos
/
Cilios
Límite:
Animals
/
Humans
Idioma:
En
Revista:
J Cell Sci
Año:
2024
Tipo del documento:
Article
País de afiliación:
Estados Unidos