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A Simple Protocol for Imaging Floral Tissues of Arabidopsis with Confocal Microscopy

  • Andrea Gómez-Felipe
  • Stefan de FolterEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1932)

Abstract

We present a simple protocol to image floral tissues with confocal laser scanning microscopy (CLSM). Recently, new imaging techniques have emerged that improve the image quality of plant tissues. In this protocol, as an example, we focus on the fluorescence detection of the miRNA MIR164c precursor. Briefly, the method involves tissue clearing, cell wall staining, and the visualization of fluorescence in tissues in young floral buds of Arabidopsis with CLSM with the use of water dipping lenses.

Key words

miRNA MIR164 Confocal microscopy ClearSee Staining Expression Fluorescence RS2200 

Notes

Acknowledgments

We thank the ABRC stock center for seeds of the pMIR164c::3xVENUS-N7 reporter line (CS65834). We thank the Mexican National Council of Science and Technology (CONACyT) for a PhD fellowship to AGF. Work in the SDF laboratory was financed by the CONACyT grants CB-2012-177739 and FC-2015-2/1061. SDF acknowledges the infrastructure grant INFR-2015-253504 to acquire the confocal microscope and acknowledges support of the European Union H2020-MSCA-RISE-2015 project ExpoSEED (grant no. 691109).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Unidad de Genómica Avanzada, Laboratorio Nacional de Genómica para la Biodiversidad (LANGEBIO)Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional (CINVESTAV-IPN)IrapuatoMexico

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