Unverified paper record
In Vivo Light Sheet Fluorescence Microscopy of Calcium Oscillations in Arabidopsis thaliana.
Methods in molecular biology (Clifton, N.J.) · 1 Jan 2019 · 10.1007/978-1-4939-9018-4_8
Abstract
Calcium imaging in plants requires a high-resolution microscope, able to perform volumetric acquisition in a few seconds, inducing as low photobleaching and phototoxicity as possible to the sample. Light sheet fluorescence microscopy offers these capabilities, with the further chance to mount the sample in vertical position, mimicking the plant's growth and physiological conditions.A protocol for plant preparation and mounting in a light sheet microscope is presented. First, the growth of Arabidopsis thaliana in a sample holder compatible with light sheet microscopy is described. Then, the requirements for sample alignment and image acquisition are detailed. Finally, the image processing steps to analyze calcium oscillations are discussed, with particular emphasis on ratiometric calcium imaging in Arabidopsis root hairs.
Plant phenotyping relevance
植物のカルシウム振動を取得・解析するライトシート蛍光イメージングの試料調製、撮像、画像処理プロトコルが中心であり、植物の生理状態を測定する手法論文である。
abstractCalcium imaging in plants requires a high-resolution microscope, able to perform volumetric acquisition in a few seconds, inducing as low photobleaching and phototoxicity as possible to the sample.
abstractA protocol for plant preparation and mounting in a light sheet microscope is presented.
abstractFinally, the image processing steps to analyze calcium oscillations are discussed, with particular emphasis on ratiometric calcium imaging in Arabidopsis root hairs.
Code and data availability
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