Intravital imaging of fluorescent markers and FRET probes by DNA tattooing

Abstract

BACKGROUND Advances in fluorescence microscopy and mouse transgenesis have made it possible to image molecular events in living animals. However, the generation of transgenic mice is a lengthy process and intravital imaging requires specialized knowledge and equipment. Here, we report a rapid and undemanding intravital imaging method using generally available equipment. RESULTS By DNA tattooing we transfect keratinocytes of living mice with DNA encoding fluorescent biosensors. Subsequently, the behavior of individual cells expressing these biosensors can be visualized within hours and using conventional microscopy equipment. Using this "instant transgenic" model in combination with a corrected coordinate system, we followed the in vivo behavior of individual cells expressing either FRET- or location-based biosensors for several days. The utility of this approach was demonstrated by assessment of in vivo caspase-3 activation upon induction of apoptosis. CONCLUSION This "instant skin transgenic" model can be used to follow the in vivo behavior of individual cells expressing either FRET- or location-based probes for several days after tattooing and provides a rapid and inexpensive method for intravital imaging in murine skin.

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Cite this paper

@article{Bins2007IntravitalIO, title={Intravital imaging of fluorescent markers and FRET probes by DNA tattooing}, author={Adriaan D Bins and Jacco van Rheenen and Kees Jalink and Jonathan R. Halstead and Nullin Divecha and David M . Spencer and John B A G Haanen and Ton NM Schumacher}, journal={BMC Biotechnology}, year={2007}, volume={7}, pages={2 - 2} }