Full developmental potential of mammalian preimplantation embryos is maintained after imaging using a spinning-disk confocal microscope.
Fluorescent live imaging of cells and embryos at subcellular resolution poses significant challenges for biologists due to morbidity and mortality ensuing from phototoxicity. Here we report the use of a spinning-disk confocal microscope to image mouse and bovine preimplantation embryos without impairing their developmental potential. We also present data indicating that this imaging technique does not affect the functionality of subcellular components as assessed by reactive oxygen species (ROS) production, caspase activity, and DNA integrity. Spinning-disk confocal microscopy was also useful in determining cell number and allocation in transgenic bovine blastocysts. We conclude that this imaging method is suitable for monitoring preimplantation embryos.
Ross
Pablo Juan
PJ
The Cellular Reprogramming Laboratory, Michigan State University, East Lansing, MI, USA.
Perez
Gloria Ines
GI
Ko
Tak
T
Yoo
Myung Sik
MS
Cibelli
Jose Bernardo
JB
D000818:Animals; D001755:Blastocyst / Q000502:physiology* / Q000648:ultrastructure*; D002417:Cattle; D002467:Cell Nucleus / Q000648:ultrastructure; D047108:Embryonic Development; D005260:Female; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods*; D017382:Reactive Oxygen Species
D017382:Reactive Oxygen Species