Dynamic <it>in vivo </it>imaging and cell tracking using a histone fluorescent protein fusion in mice

• Main Authors: Papaioannou Virginia E, Hadjantonakis Anna-Katerina
• Format: Article
• Language: English
• Published: BMC 2004-12-01
• Series: BMC Biotechnology
• Online Access: http://www.biomedcentral.com/1472-6750/4/33

<p>Abstract</p> <p>Background</p> <p>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.</p> <p>Results</p> <p>We have used embryonic stem cell-mediated transgenesis to label cells at sub-cellular resolution <it>in vivo</it>, 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.</p> <p>Conclusions</p> <p>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 <it>in situ</it>. 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 <it>in vivo </it>cell behavior and cell fate.</p>