3D time-lapse microscopy paired with endpoint lineage analysis in mouse blastocysts

STAR Protoc. 2021 Apr 8;2(2):100446. doi: 10.1016/j.xpro.2021.100446. eCollection 2021 Jun 18.


Determining how signaling dynamics relate to gene expression and cell fate is essential to understanding multicellular development. We present a unified live imaging and lineage analysis method that allows integrated analysis of both techniques in the same mouse embryos. This protocol describes the embryo isolation, confocal imaging, immunofluorescence, and in silico alignment required to connect time-lapse and endpoint measurements. By utilizing different biosensors and fixed readouts, this method allows interrogation of signaling dynamics that specify cell fates in developing embryos. For complete details on the use and execution of this protocol, please refer to Pokrass et al. (2020).

Keywords: Cell differentiation; Microscopy; Signal transduction; Single cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Blastocyst* / cytology
  • Blastocyst* / metabolism
  • Blastocyst* / physiology
  • Cells, Cultured
  • Female
  • Imaging, Three-Dimensional
  • Male
  • Mice
  • Microscopy, Confocal / methods*
  • Molecular Probe Techniques*
  • Signal Transduction / physiology
  • Time-Lapse Imaging / methods*