How genes are expressed is a fundamental aspect of biology. An unmet challenge is to examine the three-dimensional gene expression patterns in intact organisms without disruption. Current technologies to follow gene expression in vivo either fail to penetrate deeply enough or provide too poor of a resolution for many basic research questions. Optical coherence microscopy (OCM) allows non-destructive, repetitive, quantitative, in vivo three-dimensional visualization of tissues and organs in intact organisms. Previous proof-of-concept experiments demonstrated that OCM could be used to non-destructively image in vivo, three-dimensional gene expression from a strong promoter using a naturally occurring polymer, PHB (poly-D-(-)-3-hydroxybutyrate) as the reporter gene. The goal of this project is to investigate the sensitivity of this method. Transgenic plants will be generated in which the gene reporter (PHB) is expressed at different levels and at different locations. These plants will then be analyzed using OCM to visualize the reporter gene product, and computer vision methods will be applied with the goal of determining the sensitivity of the method.
Science, at its best, creates the future. A key part of how science "creates" the future is through discovery and understanding while educating others. Part of the support for the project will be for retention of a minority undergraduate in the Medford laboratory. In addition, partial support will be provided to two graduate students. To enhance the local impact of the work, an outreach activity to a local K-12 school is also planned. To enhance the wider educational impact of the work, information from the technology will be incorporated into classroom instruction. To enhance the wider scientific impact, presentations at professional meetings and publications in peer-reviewed journals are planned. This work will apply knowledge of engineers and computer vision specialists to develop the next-generation of gene analysis instruments for biologists. As such, this project is firmly positioned to significantly enhance scientific and technological understanding.
