Original video: https://www.youtube.com/watch?v=W_U6CAKjFHE
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, it is presented a detailed 3D visualization of mature flowers, and fruits, with a focus on small structures up to 60 mm tall and 45 mm deep. Using M-OPT imaging, we highlight a pea pod (Pisum sativum) in red through transmission, accompanied by two emission channels: one using a TXR filter in green and another with a GFP1 filter in blue. The scan resolution is set at 62.5 µm per voxel, offering high precision in visualizing the ovary (oa) and seeds (se), as well as air bubbles (red) between peas. Additionally, we explore a passion flower (Passiflora caerulea) using fluorescent emission M-OPT, specifically employing a GFP1 filter to capture details such as the coronal filaments (cf), ovary (oa), and petal (pt) with the same 62.5 µm per voxel resolution. The vibrant contrast between transmission and emission channels provides a clear, layered perspective of these intricate botanical structures.
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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