Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron Microscopy

In plants, the first interaction between the pollen grain and the epidermal cells of the stigma is crucial for successful reproduction. When the pollen is accepted, it germinates, producing a tube that transports the two sperm cells to the ovules for fertilization. Confocal microscopy has been used...

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Main Authors: Lucie Riglet, Isabelle Fobis-Loisy
Format: Article
Language:English
Published: Bio-protocol LLC 2024-11-01
Series:Bio-Protocol
Online Access:https://bio-protocol.org/en/bpdetail?id=5110&type=0
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author Lucie Riglet
Isabelle Fobis-Loisy
author_facet Lucie Riglet
Isabelle Fobis-Loisy
author_sort Lucie Riglet
collection DOAJ
description In plants, the first interaction between the pollen grain and the epidermal cells of the stigma is crucial for successful reproduction. When the pollen is accepted, it germinates, producing a tube that transports the two sperm cells to the ovules for fertilization. Confocal microscopy has been used to characterize the behavior of stigmatic cells post-pollination [1], but it is time-consuming since it requires the development of a range of fluorescent marker lines. Here, we propose a quick, high-resolution imaging protocol using tabletop scanning electron microscopy. This technique does not require prior sample fixation or fluorescent marker lines. It effectively captures pollen grain behavior from early hydration (a few minutes after pollination) to pollen tube growth within the stigma (1 h after pollination) and is particularly efficient for tracking pollen tube paths.
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spelling doaj-art-5347b51f5a1f453ab2e9fd40b7ca29c12025-08-20T02:35:30ZengBio-protocol LLCBio-Protocol2331-83252024-11-01142210.21769/BioProtoc.5110Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron MicroscopyLucie Riglet0Isabelle Fobis-Loisy1The Sainsbury Laboratory, University of Cambridge, 47 Bateman Street, Cambridge, UKLaboratoire Reproduction et Développement des Plantes, Univ Lyon, ENS de Lyon, UCB Lyon1, CNRS, INRAE, Lyon, FranceLaboratoire Reproduction et Développement des Plantes, Univ Lyon, ENS de Lyon, UCB Lyon1, CNRS, INRAE, Lyon, FranceIn plants, the first interaction between the pollen grain and the epidermal cells of the stigma is crucial for successful reproduction. When the pollen is accepted, it germinates, producing a tube that transports the two sperm cells to the ovules for fertilization. Confocal microscopy has been used to characterize the behavior of stigmatic cells post-pollination [1], but it is time-consuming since it requires the development of a range of fluorescent marker lines. Here, we propose a quick, high-resolution imaging protocol using tabletop scanning electron microscopy. This technique does not require prior sample fixation or fluorescent marker lines. It effectively captures pollen grain behavior from early hydration (a few minutes after pollination) to pollen tube growth within the stigma (1 h after pollination) and is particularly efficient for tracking pollen tube paths.https://bio-protocol.org/en/bpdetail?id=5110&type=0
spellingShingle Lucie Riglet
Isabelle Fobis-Loisy
Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron Microscopy
Bio-Protocol
title Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron Microscopy
title_full Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron Microscopy
title_fullStr Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron Microscopy
title_full_unstemmed Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron Microscopy
title_short Fast and High-Resolution Imaging of Pollinated Stigmatic Cells by Tabletop Scanning Electron Microscopy
title_sort fast and high resolution imaging of pollinated stigmatic cells by tabletop scanning electron microscopy
url https://bio-protocol.org/en/bpdetail?id=5110&type=0
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