Abstract
BackgroundIn vivo imaging and quantification of fluorescent reporter molecules is increasingly useful in biomedical research. For example, tracking animal movement in 3D with simultaneous quantification of fluorescent transgenic reporters allows for correlations between behavior, aging and gene expression. However implementation has been hindered in the past by the complexity of operating the systems.ResultsWe report significant technical improvements and user-friendly software (called FluoreScore) that enables tracking of 3D movement and the dynamics of gene expression in adult Drosophila, using two cameras and recorded GFP videos. Expression of a transgenic construct encoding eGFP was induced in free-moving adult flies using the Gene-Switch system and RU486 drug feeding. The time course of induction of eGFP expression was readily quantified from internal tissues including central nervous tissue.ConclusionsFluoreScore should facilitate a variety of future studies involving quantification of movement behaviors and fluorescent molecules in free-moving animals.
Highlights
In vivo monitoring of fluorescent and luminescent reporter molecules is proving increasingly useful in the study of cell behavior and physiology, including stem cell dynamics, cancer, neurobiology and aging [1,2,3,4]
We have previously reported methods that allow for 3D video tracking of Drosophila melanogaster movement behaviors [6,7], while simultaneously quantifying gene expression using fluorescent reporter proteins [8,9]
In addition to tracking 3D movement, these methods allowed for quantification of GFP reporter induction in internal tissues in response to drug feeding and transcriptional activation in free-moving flies
Summary
In vivo monitoring of fluorescent and luminescent reporter molecules is proving increasingly useful in the study of cell behavior and physiology, including stem cell dynamics, cancer, neurobiology and aging [1,2,3,4]. Transgenic constructs that drive expression of fluorescent molecules such as GFP allow for non-invasive and longitudinal assays of gene expression [5]. We have previously reported methods that allow for 3D video tracking of Drosophila melanogaster movement behaviors [6,7], while simultaneously quantifying gene expression using fluorescent reporter proteins [8,9]. In vivo imaging and quantification of fluorescent reporter molecules is increasingly useful in biomedical research. Tracking animal movement in 3D with simultaneous quantification of fluorescent transgenic reporters allows for correlations between behavior, aging and gene expression. Implementation has been hindered in the past by the complexity of operating the systems
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