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Automated Image Acquisition and Morphological Analysis of Cell Growth Mutants in Physcomitrella patens

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Book cover Plant Cell Morphogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1992))

Abstract

This protocol describes the automated imaging and a quantitative analysis of the morphology of small plants from the moss Physcomitrella patens. This method can be used for the analysis of growth phenotypes produced by transient RNA interference or for the analysis of stable mutant plants. Furthermore, we describe how to acquire higher resolution images via the acquisition of a collection of multiple overlapping tiles from the same image. Information is presented to guide the investigator in the choice of vectors and basic conditions to perform transient RNA interference in moss. Detailed directions and examples for fluorescence image acquisition of small regenerating moss plants are provided. Instructions for stitching image tiles and for using an ImageJ-based macro for the quantitative morphological analysis of moss plants are also provided.

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Acknowledgments

This work was supported in part by NSF (IOS 1002837 and MCB 1253444 to L.V.). We thank Erin Agar, Jennifer Garbarino, and Graham Burkart for comments on the manuscript and to Luisanna Paulino for sample images. We also thank other members of the Vidali and Bezanilla laboratories for their support.

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Authors and Affiliations

  1. Department of Biology and Biotechnology, Worcester Polytechnic Institute, Worcester, MA, USA

    Giulia Galotto, Jeffrey P. Bibeau & Luis Vidali

Authors
  1. Giulia Galotto
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  2. Jeffrey P. Bibeau
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  3. Luis Vidali
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Corresponding author

Correspondence to Luis Vidali .

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Editors and Affiliations

  1. Department of Experimental Plant Biology, Charles University, Prague, Czech Republic

    Fatima Cvrčková

  2. Department of Experimental Plant Biology, Charles University, Prague, Czech Republic

    Viktor Žárský

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Galotto, G., Bibeau, J.P., Vidali, L. (2019). Automated Image Acquisition and Morphological Analysis of Cell Growth Mutants in Physcomitrella patens. In: Cvrčková, F., Žárský, V. (eds) Plant Cell Morphogenesis. Methods in Molecular Biology, vol 1992. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9469-4_20

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  • DOI: https://doi.org/10.1007/978-1-4939-9469-4_20

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9468-7

  • Online ISBN: 978-1-4939-9469-4

  • eBook Packages: Springer Protocols

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