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. 2018 Oct 13;14:89. doi: 10.1186/s13007-018-0356-x

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© The Author(s) 2018

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. 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.

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Fig. 4 — Cryo-observation of apple a, c, e and grape b, d, f tissues infused by Fluo-3 (calcium), PBFI (potassium), DPBA (flavonoid) dyes and cryo-fixed in cooled isopentane. Fluorescence observations in fresh apple fruit (g, i, k) and grape berry (h, j, l) sections directly stained by Fluo-3, PBFI and DPBA. C: Cuticle, CW: Cell Wall. Scale bar: 50 µm for calcium probe and 100 µm for potassium and flavonoid probes. Laser excitation and gain are indicated to stress the difference in fluorescence intensity recovered between fresh and frozen samples. The applied fluorescence intensity of the cuticle is due in part to autofluorescence related to the presence of phenolic compounds