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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...

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Autores principales: Welling, Maaike, Mohr, Manuel Alexander, Ponti, Aaron, Rullan Sabater, Lluc, Boni, Andrea, Kawamura, Yumiko K, Liberali, Prisca, Peters, Antoine HFM, Pelczar, Pawel, Pantazis, Periklis
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2019
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.
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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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