Optimization of cell spreading and image quality for the study of chromosomes in plant tissues

Laksana Kantama, Erik Wijnker, Hans de Jong*

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

High-quality chromosome images of mitotic and meiotic cell divisions in plant tissues are inextricably connected with the technical control of cell spread preparations. Superb chromosome slides are the best for studying chromosome morphology and making karyotypes; they also are the best start for a successful fluorescent in situ hybridization experiment. In this study, we describe the essentials for fixation, enzymatic digestion, squash, spread, and dropping protocols for clean and well-differentiated nuclei and chromosome complements. In addition, we focus on the use of standard whole image processing for best sharpness, brightness and contrast adjustments, differentiation of heterochromatin/euchromatin, and high dynamic range imaging of big chromosomes. We also explain how to combine transparent layers or spot channels of different fluorescent images for making publication quality, full color photos.

Original languageEnglish
Title of host publicationPlant Germline Development
Subtitle of host publicationMethods and Protocols
EditorsAnja Schmidt
PublisherHumana Press Inc.
Pages141-158
Volume1669
ISBN (Electronic)9781493972869
ISBN (Print)9781493972852
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1669
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Cell spreading protocol
  • Chromosome images
  • Chromosome technique
  • Image processing microphotographs

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    Kantama, L., Wijnker, E., & de Jong, H. (2017). Optimization of cell spreading and image quality for the study of chromosomes in plant tissues. In A. Schmidt (Ed.), Plant Germline Development: Methods and Protocols (Vol. 1669, pp. 141-158). (Methods in Molecular Biology; Vol. 1669). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7286-9_12