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      A device for the controlled cooling and freezing of excised plant specimens during magnetic resonance imaging

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          Abstract

          Background

          Investigating plant mechanisms to tolerate freezing temperatures is critical to developing crops with superior cold hardiness. However, the lack of imaging methods that allow the visualization of freezing events in complex plant tissues remains a key limitation. Magnetic resonance imaging (MRI) has been successfully used to study many different plant models, including the study of in vivo changes during freezing. However, despite its benefits and past successes, the use of MRI in plant sciences remains low, likely due to limited access, high costs, and associated engineering challenges, such as keeping samples frozen for cold hardiness studies. To address this latter need, a novel device for keeping plant specimens at freezing temperatures during MRI is described.

          Results

          The device consists of commercial and custom parts. All custom parts were 3D printed and made available as open source to increase accessibility to research groups who wish to reproduce or iterate on this work. Calibration tests documented that, upon temperature equilibration for a given experimental temperature, conditions between the circulating coolant bath and inside the device seated within the bore of the magnet varied by less than 0.1 °C. The device was tested on plant material by imaging buds from Vaccinium macrocarpon in a small animal MRI system, at four temperatures, 20 °C, − 7 °C, − 14 °C, and −  21 °C. Results were compared to those obtained by independent controlled freezing test (CFT) evaluations. Non-damaging freezing events in inner bud structures were detected from the imaging data collected using this device, phenomena that are undetectable using CFT.

          Conclusions

          The use of this novel cooling and freezing device in conjunction with MRI facilitated the detection of freezing events in intact plant tissues through the observation of the presence and absence of water in liquid state. The device represents an important addition to plant imaging tools currently available to researchers. Furthermore, its open-source and customizable design ensures that it will be accessible to a wide range of researchers and applications.

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          Most cited references16

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          NIH Image to ImageJ: 25 years of image analysis

          For the past twenty five years the NIH family of imaging software, NIH Image and ImageJ have been pioneers as open tools for scientific image analysis. We discuss the origins, challenges and solutions of these two programs, and how their history can serve to advise and inform other software projects.
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            Surveying the plant's world by magnetic resonance imaging.

            Understanding the way in which plants develop, grow and interact with their environment requires tools capable of a high degree of both spatial and temporal resolution. Magnetic resonance imaging (MRI), a technique which is able to visualize internal structures and metabolites, has the great virtue that it is non-invasive and therefore has the potential to monitor physiological processes occurring in vivo. The major aim of this review is to attract plant biologists to MRI by explaining its advantages and wide range of possible applications for solving outstanding issues in plant science. We discuss the challenges and opportunities of MRI in the study of plant physiology and development, plant-environment interactions, biodiversity, gene functions and metabolism. Overall, it is our view that the potential benefit of harnessing MRI for plant research purposes is hard to overrate. © 2012 The Authors. The Plant Journal © 2012 Blackwell Publishing Ltd.
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              Changes in aquaporin gene expression and magnetic resonance imaging of water status in peach tree flower buds during dormancy.

              The movement of cellular water accompanies changes in growth within dormant buds. To further understand this process, accumulation of tonoplast deltaTIP1 and plasma membrane PIP2 aquaporin transcripts was measured by quantitative reverse transcriptase-polymerase chain reaction and the water dynamics in dormant peach (Prunus persica L.) flower buds was studied by magnetic resonance imaging. Proton density (PD), spin-spin relaxation time (T(2)) and apparent diffusion coefficient (ADC) were used to observe water dynamics during dormancy. The expression of deltaTIP1 and PIP2 aquaporins, PD and T(2) in the upper part of the bud including primordia, in the basal part of the bud and the bud trace increased earlier in the low-chill cultivar 'Coral' than in the high-chill cultivar 'Kansuke Hakuto,' reflecting the difference in timing for the end of endodormancy in the two cultivars. deltaTIP1 mRNA accumulated mainly in the basal part of the bud, whereas PIP2 mRNA was detected mainly in the upper part. These findings may reflect the activation of inter- and intracell communication through membrane transport properties of aquaporins resulting in a gradual increase in water content to that required for bud activity at the end of endodormancy. An apparent decrease in the expression of deltaTIP1 and PIP2 mRNAs was, however, observed in late winter in some portions of the buds of both cultivars just before sprouting.
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                Author and article information

                Contributors
                atucha@wisc.edu
                Journal
                Plant Methods
                Plant Methods
                Plant Methods
                BioMed Central (London )
                1746-4811
                13 April 2021
                13 April 2021
                2021
                : 17
                : 41
                Affiliations
                [1 ]GRID grid.14003.36, ISNI 0000 0001 2167 3675, Department of Horticulture, , University of Wisconsin-Madison, ; 1575 Linden Dr., Madison, WI 53706 USA
                [2 ]GRID grid.14003.36, ISNI 0000 0001 2167 3675, Medical Engineering Group, , Morgridge Institute for Research, ; 330 N Orchard St, Madison, WI 53706 USA
                [3 ]GRID grid.14003.36, ISNI 0000 0001 2167 3675, Laboratory for Optical and Computational Instrumentation (LOCI), , University of Wisconsin-Madison, ; 1675 Observatory Dr., Madison, WI 53706 USA
                [4 ]GRID grid.14003.36, ISNI 0000 0001 2167 3675, Department of Medical Physics, , University of Wisconsin-Madison, ; 1111 Highland Ave., Madison, WI 53705 USA
                [5 ]GRID grid.14003.36, ISNI 0000 0001 2167 3675, Department of Biomedical Engineering, , University of Wisconsin-Madison, ; 1415 Engineering Dr., Madison, WI 53706 USA
                Author information
                http://orcid.org/0000-0002-8921-5096
                Article
                743
                10.1186/s13007-021-00743-4
                8045372
                a3a41e6b-19f6-47ef-9d93-6656b641e5d6
                © The Author(s) 2021

                Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

                History
                : 19 November 2020
                : 4 April 2021
                Categories
                Methodology
                Custom metadata
                © The Author(s) 2021

                Plant science & Botany
                mri,3d printing,dormant bud,freezing resistance,plant cold hardiness
                Plant science & Botany
                mri, 3d printing, dormant bud, freezing resistance, plant cold hardiness

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