Original video: https://www.youtube.com/watch?v=SC04kxvFqxE
Optical projection tomography (OPT) is a well-established method for visualising gene activity in plants and animals. However, a limitation of conventional OPT is that the specimen upper size limit precludes its application to larger structures. To address this problem we constructed a macro version called Macro OPT (M-OPT). We apply M-OPT to 3D live imaging of gene activity in growing whole plants and to visualise structural morphology in large optically cleared plant and insect specimens up to 60 mm tall and 45 mm deep. We also show how M-OPT can be used to image gene expression domains in 3D within fixed tissue and to visualise gene activity in 3D in clones of growing young whole Arabidopsis plants. A further application of M-OPT is to visualise plant-insect interactions. Thus M-OPT provides an effective 3D imaging platform that allows the study of gene activity, internal plant structures and plant-insect interactions at a macroscopic scale.
In this video, we showcase a clipped volume view of a daffodil flower (Narcissus poeticus) using M-OPT imaging. The transmission channel is displayed in red, while two emission channels are combined—using a GFP1 filter in green and a TXR filter in blue—to provide a comprehensive visualization. The scan resolution is set at 40 µm per voxel, offering intricate details of the flower’s structure.
Next, we present a volume view of an immature Venus flytrap (Dionaea muscipula), focusing on the arrangement of veins within the valve and the interlocking cilia along the valve margins. The imaging includes a red transmission channel and a GFP1 emission channel in green, with a scan resolution of 38.2 µm per voxel. These visualizations highlight the complex structural details of both plants, offering a unique perspective on their anatomy.
Keywords: 3D, development, gene activity, imaging, macroscopic, macro optical projection tomography, M-OPT, plant-insect interactions, tomography, whole plant.
Citation: Karen J. I. Lee, Grant M. Calder, Christopher R. Hindle, Jacob L. Newman, Simon N. Robinson, Jerome J. H. Y. Avondo, Enrico S. Coen, Macro optical projection tomography for large scale 3D imaging of plant structures and gene activity, Journal of Experimental Botany, Volume 68, Issue 3, 1 January 2017, Pages 527–538, https://doi.org/10.1093/jxb/erw452
Published: 26 December 2016
Attribution 4.0 International — CC BY 4.0 - Creative Commons
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