Quantitative semi-automated analysis of morphogenesis with single-cell resolution in complex embryos.
A quantitative understanding of tissue morphogenesis requires description of the movements of individual cells in space and over time. In transparent embryos, such as C. elegans, fluorescently labeled nuclei can be imaged in three-dimensional time-lapse (4D) movies and automatically tracked through early cleavage divisions up to ~350 nuclei. A similar analysis of later stages of C. elegans development has been challenging owing to the increased error rates of automated tracking of large numbers of densely packed nuclei. We present Nucleitracker4D, a freely available software solution for tracking nuclei in complex embryos that integrates automated tracking of nuclei in local searches with manual curation. Using these methods, we have been able to track >99% of all nuclei generated in the C. elegans embryo. Our analysis reveals that ventral enclosure of the epidermis is accompanied by complex coordinated migration of the neuronal substrate. We can efficiently track large numbers of migrating nuclei in 4D movies of zebrafish cardiac morphogenesis, suggesting that this approach is generally useful in situations in which the number, packing or dynamics of nuclei present challenges for automated tracking.
Division of Biological Sciences, Section of Cell and Developmental Biology, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093,  USA.
Claudiu A
CA
Giurumescu
Sukryool
S
Kang
Thomas A
TA
Planchon
Eric
E
Betzig
Joshua
J
Bloomekatz
Deborah
D
Yelon
Pamela
P
Cosman
Andrew D
AD
Chisholm
D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology*; D002454:Cell Differentiation; D002455:Cell Division; D002465:Cell Movement; D002467:Cell Nucleus / Q000378:metabolism; D003201:Computers; D004625:Embryo, Nonmammalian; D004817:Epidermis / Q000378:metabolism; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D009024:Morphogenesis; D059010:Single-Cell Analysis; D012984:Software; D013223:Statistics as Topic; D015027:Zebrafish / Q000196:embryology*