Detection and Automated Analysis of Single Transcripts at Subcellular Resolution in Zebrafish Embryos.
Methods Mol Biol
; 1649: 143-162, 2018.
Article
en En
| MEDLINE
| ID: mdl-29130195
ABSTRACT
Single molecule fluorescence in situ hybridization (smFISH) is a method to visualize single mRNA molecules. When combined with cellular and nuclear segmentation, transcripts can be assigned to different cellular compartments resulting in quantitative information on transcript levels at subcellular resolution. The use of smFISH in zebrafish has been limited by the lack of protocols and an automated image analysis pipeline for samples of multicellular organisms. Here we present a protocol for smFISH on zebrafish cryosections. The protocol includes a method to obtain high-quality sections of zebrafish embryos, an smFISH protocol optimized for zebrafish cryosections, and a user-friendly, automated analysis pipeline for cell segmentation and transcript detection. The software is freely available and can be used to analyze sections of any multicellular organism.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Pez Cebra
/
ARN Mensajero
/
Hibridación Fluorescente in Situ
/
Embrión no Mamífero
Tipo de estudio:
Diagnostic_studies
Límite:
Animals
Idioma:
En
Revista:
Methods Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2018
Tipo del documento:
Article
País de afiliación:
Alemania