Please note that eDoc will be permanently shut down in the first quarter of 2021!      Home News About Us Contact Contributors Disclaimer Privacy Policy Help FAQ
Home Search Quick Search Advanced Fulltext Browse Collections Persons My eDoc Session History Login Name: Password: Documentation Help Support Wiki Direct access to document ID:	Institute: MPI für molekulare Zellbiologie und Genetik     Collection: MPI-CBG Publications 2016 (archival)     Display Documents ID: 732472.0, MPI für molekulare Zellbiologie und Genetik / MPI-CBG Publications 2016 (archival) Automated detection and quantification of single RNAs at cellular resolution in zebrafish embryos. Authors: Stapel, L Carine; Lombardot, Benoit; Broaddus, Coleman; Kainmueller, Dagmar; Jug, Florian; Myers, Eugene W; Vastenhouw, Nadine Date of Publication (YYYY-MM-DD): 2016 Title of Journal: Development (Cambridge, England) Volume: 143 Issue / Number: 3 Start Page: 540 End Page: 546 Copyright: not available Audience: Experts Only Intended Educational Use: No Abstract / Description: Analysis of differential gene expression is crucial for the study of cell fate and behavior during embryonic development. However, automated methods for the sensitive detection and quantification of RNAs at cellular resolution in embryos are lacking. With the advent of single-molecule fluorescence in situ hybridization (smFISH), gene expression can be analyzed at single-molecule resolution. However, the limited availability of protocols for smFISH in embryos and the lack of efficient image analysis pipelines have hampered quantification at the (sub)cellular level in complex samples such as tissues and embryos. Here, we present a protocol for smFISH on zebrafish embryo sections in combination with an image analysis pipeline for automated transcript detection and cell segmentation. We use this strategy to quantify gene expression differences between different cell types and identify differences in subcellular transcript localization between genes. The combination of our smFISH protocol and custom-made, freely available, analysis pipeline will enable researchers to fully exploit the benefits of quantitative transcript analysis at cellular and subcellular resolution in tissues and embryos. External Publication Status: published Document Type: Article Communicated by: Thüm Affiliations: MPI für molekulare Zellbiologie und Genetik Identifiers: LOCALID:6424 The scope and number of records on eDoc is subject to the collection policies defined by each institute - see "info" button in the collection browse view.