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High fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins

Maaike Welling, View ORCID ProfileManuel Alexander Mohr, Aaron Ponti, Lluc Rullan Sabater, Andrea Boni, Prisca Liberali, Pawel Pelczar, View ORCID ProfilePeriklis Pantazis
doi: https://doi.org/10.1101/218354
Maaike Welling
1Department for Biosystems Science and Engineering (D-BSSE), Eidgenössische Technische Hochschule (ETH) Zurich, 4058 Basel, Switzerland
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Manuel Alexander Mohr
1Department for Biosystems Science and Engineering (D-BSSE), Eidgenössische Technische Hochschule (ETH) Zurich, 4058 Basel, Switzerland
2Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147, USA
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Aaron Ponti
1Department for Biosystems Science and Engineering (D-BSSE), Eidgenössische Technische Hochschule (ETH) Zurich, 4058 Basel, Switzerland
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Lluc Rullan Sabater
1Department for Biosystems Science and Engineering (D-BSSE), Eidgenössische Technische Hochschule (ETH) Zurich, 4058 Basel, Switzerland
3present address: QuintilesIMS, 4053 Basel, Switzerland
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Andrea Boni
4Friedrich Miescher Institute for Biomedical Research (FMI), 4058 Basel, Switzerland
5present address: Ecole polytechinque federale de Lausanne (EPFL), 1015 Lausanne, Switzerland
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Prisca Liberali
4Friedrich Miescher Institute for Biomedical Research (FMI), 4058 Basel, Switzerland
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Pawel Pelczar
6Center for Transgenic Models (CTM), University of Basel, 4002 Basel, Switzerland
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Periklis Pantazis
1Department for Biosystems Science and Engineering (D-BSSE), Eidgenössische Technische Hochschule (ETH) Zurich, 4058 Basel, Switzerland
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  • ORCID record for Periklis Pantazis
  • For correspondence: periklis.pantazis@bsse.ethz.ch
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Abstract

Accurate lineage reconstruction of mammalian pre-implantation development is essential for inferring the earliest cell fate decisions of mammalian development. Lineage tracing using global labeling techniques is complicated by increasing cell density and rapid embryo rotation, impeding automatic alignment and rendering accurate cell tracking of obtained four-dimensional imaging data sets highly challenging. Here, we exploit the advantageous properties of primed convertible fluorescent proteins (pr-pcFPs) to simultaneously visualize the global green and the photoconverted red population 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 and accomplish high fidelity lineage tracing combined with a reduced data size – addressing majors concerns in the field of volumetric embryo imaging.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted November 12, 2017.
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High fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins
Maaike Welling, Manuel Alexander Mohr, Aaron Ponti, Lluc Rullan Sabater, Andrea Boni, Prisca Liberali, Pawel Pelczar, Periklis Pantazis
bioRxiv 218354; doi: https://doi.org/10.1101/218354
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High fidelity lineage tracing in mouse pre-implantation embryos using primed conversion of photoconvertible proteins
Maaike Welling, Manuel Alexander Mohr, Aaron Ponti, Lluc Rullan Sabater, Andrea Boni, Prisca Liberali, Pawel Pelczar, Periklis Pantazis
bioRxiv 218354; doi: https://doi.org/10.1101/218354

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