A012 - Imaging transcription at the single cell level during zebrafish development

Maelle Bellec (maelle.bellec@mpi-bn.mpg.de)
Jeremy Dufourt (jeremy.dufourt@igmm.cnrs.fr)

Live imaging represents a powerful technique for the understanding of dynamic processes in biology, such as embryonic development. Recent advances in microscopy highly improve signal-to-noise ratios as well as spatial and temporal resolutions. Furthermore, development of new fluorescence markers allows a better quantification of protein expression and transcriptional dynamics in vivo. This workshop aims at imaging transcription in live, within whole zebrafish embryos in order to extract transcriptional dynamics. To do that, we will use the MS2/MCP system which consists of MS2 loops that upon transcription will be recognized by a MS2 Coat Protein fused to GFP. We will first show how to prepare and mount a living sample adapted for confocal microscopy. We will then explain how to make imaging settings in order to get a good signal to noise ratio and high temporal resolution with this system. We will use the newly developed Lattice Light Sheet microscope (ZEISS LLS7) to be able to image at a high time frequency.