Imaging exocytosis and recycling of synaptic-like microvesicles in astrocytes.
Cold Spring Harb Protoc
; 2014(5)2014 May 01.
Article
en En
| MEDLINE
| ID: mdl-24786509
ABSTRACT
Optical imaging techniques are well suited for following the dynamics of physiological processes in living cells. Total internal reflection fluorescence (TIRF) microscopy based on evanescent wave illumination (EWi) allows spectacular, real-time visualization of individual vesicle movements, fusions, and retrievals at the cell surface (i.e., within 100 nm of the plasma membrane). TIRF microscopy is an ideal approach for studying the properties of exocytosis and recycling in cultured astrocytes, particularly because these cells have a rather flat surface and contain secretory vesicles with sparse distribution. Among all populations of secretory vesicles, we focus here on synaptic-like microvesicles (SLMVs). We illustrate how TIRF microscopy using EWi is useful to study exocytosis and recycling of SLMVs at the single-vesicle level and, when combined with epifluorescence illumination (EPIi), can provide detailed information on the kinetics of exocytosis, endocytosis, and re-acidification at the whole-cell level.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Astrocitos
/
Exocitosis
/
Exosomas
/
Imagen Óptica
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Microscopía Fluorescente
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Cold Spring Harb Protoc
Año:
2014
Tipo del documento:
Article