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Primed Track, high-fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins
Accurate lineage reconstruction of mammalian pre-implantation development is essential for inferring the earliest cell fate decisions. Lineage tracing using global fluorescence labeling techniques is complicated by increasing cell density and rapid embryo rotation, which hampers automatic alignment...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6340703/ https://www.ncbi.nlm.nih.gov/pubmed/30663981 http://dx.doi.org/10.7554/eLife.44491 |
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author | Welling, Maaike Mohr, Manuel Alexander Ponti, Aaron Rullan Sabater, Lluc Boni, Andrea Kawamura, Yumiko K Liberali, Prisca Peters, Antoine HFM Pelczar, Pawel Pantazis, Periklis |
author_facet | Welling, Maaike Mohr, Manuel Alexander Ponti, Aaron Rullan Sabater, Lluc Boni, Andrea Kawamura, Yumiko K Liberali, Prisca Peters, Antoine HFM Pelczar, Pawel Pantazis, Periklis |
author_sort | Welling, Maaike |
collection | PubMed |
description | Accurate lineage reconstruction of mammalian pre-implantation development is essential for inferring the earliest cell fate decisions. Lineage tracing using global fluorescence labeling techniques is complicated by increasing cell density and rapid embryo rotation, which hampers automatic alignment and accurate cell tracking of obtained four-dimensional imaging data sets. Here, we exploit the advantageous properties of primed convertible fluorescent proteins (pr-pcFPs) to simultaneously visualize the global green and the photoconverted red population in order to minimize tracking uncertainties over prolonged time windows. Confined primed conversion of H2B-pr-mEosFP-labeled nuclei combined with light-sheet imaging greatly facilitates segmentation, classification, and tracking of individual nuclei from the 4-cell stage up to the blastocyst. Using green and red labels as fiducial markers, we computationally correct for rotational and translational drift, reduce overall data size, and accomplish high-fidelity lineage tracing even for increased imaging time intervals – addressing major concerns in the field of volumetric embryo imaging. |
format | Online Article Text |
id | pubmed-6340703 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-63407032019-01-24 Primed Track, high-fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins Welling, Maaike Mohr, Manuel Alexander Ponti, Aaron Rullan Sabater, Lluc Boni, Andrea Kawamura, Yumiko K Liberali, Prisca Peters, Antoine HFM Pelczar, Pawel Pantazis, Periklis eLife Developmental Biology Accurate lineage reconstruction of mammalian pre-implantation development is essential for inferring the earliest cell fate decisions. Lineage tracing using global fluorescence labeling techniques is complicated by increasing cell density and rapid embryo rotation, which hampers automatic alignment and accurate cell tracking of obtained four-dimensional imaging data sets. Here, we exploit the advantageous properties of primed convertible fluorescent proteins (pr-pcFPs) to simultaneously visualize the global green and the photoconverted red population in order to minimize tracking uncertainties over prolonged time windows. Confined primed conversion of H2B-pr-mEosFP-labeled nuclei combined with light-sheet imaging greatly facilitates segmentation, classification, and tracking of individual nuclei from the 4-cell stage up to the blastocyst. Using green and red labels as fiducial markers, we computationally correct for rotational and translational drift, reduce overall data size, and accomplish high-fidelity lineage tracing even for increased imaging time intervals – addressing major concerns in the field of volumetric embryo imaging. eLife Sciences Publications, Ltd 2019-01-21 /pmc/articles/PMC6340703/ /pubmed/30663981 http://dx.doi.org/10.7554/eLife.44491 Text en © 2019, Welling et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Developmental Biology Welling, Maaike Mohr, Manuel Alexander Ponti, Aaron Rullan Sabater, Lluc Boni, Andrea Kawamura, Yumiko K Liberali, Prisca Peters, Antoine HFM Pelczar, Pawel Pantazis, Periklis Primed Track, high-fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins |
title | Primed Track, high-fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins |
title_full | Primed Track, high-fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins |
title_fullStr | Primed Track, high-fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins |
title_full_unstemmed | Primed Track, high-fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins |
title_short | Primed Track, high-fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins |
title_sort | primed track, high-fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins |
topic | Developmental Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6340703/ https://www.ncbi.nlm.nih.gov/pubmed/30663981 http://dx.doi.org/10.7554/eLife.44491 |
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