Live Imaging of mRNA Transcription in Drosophila Embryos
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Live imaging has been used in recent years for the understanding of dynamic processes in biology, such as embryo development. This was made possible by a combination of advancements in microscopy, leading to improved signal-to-noise ratios and better spatial and temporal resolutions, and by the development of new fluorescence markers, allowing for the quantification of protein expression and transcriptional dynamics in vivo. Here we describe a general protocol, which can be used in standard confocal microscopes to image early Drosophila melanogaster embryos, in order to learn about the transcriptional dynamics of a fluorescently labeled RNA.
Key wordsMS2 system Live imaging Confocal microscopy RNA Embryo
The authors thank Patricia Le Baccon and the Imaging Facility PICT-IBiSA of the Institut Curie. This work was supported by a PSL IDEX REFLEX Grant for Mesoscopic Biology (ND, AMW, MC), an Ontario Trillium Scholarship for International Students (CAPR), a Mitacs Global Link Scholarship (CAPR) and an Internal Curie Institute Scholarship (CAPR), ARC PJA20151203341 (ND), a Mayent Rothschild sabbatical Grant from the Curie Institute (CF) and an NSERC discovery grant RGPIN/06362-15 (CF), a Marie Curie MCCIG grant No. 303561 (AMW), ANR-11-LABX-0044 DEEP Labex (ND), ANR- 11-BSV2-0024 Axomorph (ND and AMW) and PSL ANR-10-IDEX-0001-02. Cécile Fradin and Nathalie Dostatni contributed equally to this work. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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