Imaging cellular and molecular dynamics in live embryos using fluorescent proteins.
With the live imaging of embryos, the dynamics of developmental processes, such as tissue remodeling, cell morphogenesis, and, campus protein dynamics can be observed and quantified. This has greatly improved the mechanistic understanding of biological processes. Here we describe how embryos can be prepared for imaging mainly, but not only, fluorescent proteins and probes. This chapter is a users' guide that addresses the following aspects of fluorescent embryo imaging: (1) How to handle and prepare embryos for live microscopy. (2) What microscopic setups are available for embryo imaging and what should they be used for. (3) How to practically use fluorescent imaging setups depending on the experimental context: large-scale imaging of multiple embryos, high-resolution four-dimensional imaging of single embryos, studies of protein dynamics, and so on. (4) Finally, we focus on pitfalls and how to overcome a variety of possible problems encountered during live imaging.
Cavey
Matthieu
M
IBDML, campus de Luminy, Marseille, France.
Lecuit
Thomas
T
D000818:Animals; D015820:Cadherins / Q000378:metabolism; D015509:Developmental Biology / Q000379:methods*; D004331:Drosophila melanogaster; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D007834:Lasers; D008027:Light; D008164:Luminescent Proteins / Q000378:metabolism*; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D038761:Photobleaching; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013997:Time Factors
D015820:Cadherins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins