Academic Commons

Articles

Dynamic in vivo imaging and cell tracking using a histone fluorescent protein fusion in mice

Hadjantonakis, Anna-Katerina; Papaioannou, Virginia

Advances in optical imaging modalities and the continued evolution of genetically-encoded fluorescent proteins are coming together to facilitate the study of cell behavior at high resolution in living organisms. As a result, imaging using autofluorescent protein reporters is gaining popularity in mouse transgenic and targeted mutagenesis applications. We have used embryonic stem cell-mediated transgenesis to label cells at sub-cellular resolution in vivo, and to evaluate fusion of a human histone protein to green fluorescent protein for ubiquitous fluorescent labeling of nucleosomes in mice. To this end we have generated embryonic stem cells and a corresponding strain of mice that is viable and fertile and exhibits widespread chromatin-localized reporter expression. High levels of transgene expression are maintained in a constitutive manner. Viability and fertility of homozygous transgenic animals demonstrates that this reporter is developmentally neutral and does not interfere with mitosis or meiosis. Using various optical imaging modalities including wide-field, spinning disc confocal, and laser scanning confocal and multiphoton excitation microscopy, we can identify cells in various stages of the cell cycle. We can identify cells in interphase, cells undergoing mitosis or cell death. We demonstrate that this histone fusion reporter allows the direct visualization of active chromatin in situ. Since this reporter segments three-dimensional space, it permits the visualization of individual cells within a population, and so facilitates tracking cell position over time. It is therefore attractive for use in multidimensional studies of in vivo cell behavior and cell fate.

Files

  • thumnail for 1472-6750-4-33-S8.MOV 1472-6750-4-33-S8.MOV binary/octet-stream 4.82 MB Download File
  • thumnail for 1472-6750-4-33-S9.MOV 1472-6750-4-33-S9.MOV binary/octet-stream 14.3 MB Download File
  • thumnail for 1472-6750-4-33-S10.MOV 1472-6750-4-33-S10.MOV binary/octet-stream 12.9 MB Download File
  • thumnail for 1472-6750-4-33-S11.MOV 1472-6750-4-33-S11.MOV binary/octet-stream 15.9 MB Download File
  • thumnail for 1472-6750-4-33-S3.MOV 1472-6750-4-33-S3.MOV binary/octet-stream 2.96 MB Download File
  • thumnail for 1472-6750-4-33-S5.MOV 1472-6750-4-33-S5.MOV binary/octet-stream 12.6 MB Download File
  • thumnail for 1472-6750-4-33-S4.MOV 1472-6750-4-33-S4.MOV binary/octet-stream 12.7 MB Download File
  • thumnail for 1472-6750-4-33-S6.AVI 1472-6750-4-33-S6.AVI binary/octet-stream 8.38 MB Download File
  • thumnail for 1472-6750-4-33-S7.MOV 1472-6750-4-33-S7.MOV binary/octet-stream 45.9 MB Download File
  • thumnail for eea357ff9fd4eed1b72617756775524f.zip eea357ff9fd4eed1b72617756775524f.zip binary/octet-stream 106 MB Download File

Also Published In

Title
BMC Biotechnology
DOI
https://doi.org/10.1186/1472-6750-4-33

More About This Work

Academic Units
Genetics and Development
Publisher
BioMed Central
Published Here
September 8, 2014
Academic Commons provides global access to research and scholarship produced at Columbia University, Barnard College, Teachers College, Union Theological Seminary and Jewish Theological Seminary. Academic Commons is managed by the Columbia University Libraries.