Improved Method for Critical Comparison of Cell Cycle Data of Asynchronously Dividing and Synchronized Cell Cultures of Nicotiana tabacum

  • Pfosser M
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Abstract

An improved immunocytochemical procedure for the detection of incorporated bromodeoxyuridine was applied to plant cells to study the cell cycle of Nicotiana tabacum suspension cultures. The results corresponded very well with cell cycle data obtained by flow cytometry of 4′ ,6-diamidino-2-phenylindole (DAPI)-stained nuclei released from asynchronously dividing cells. After synchronization with hydroxyurea, flow cytometry alone was insufficient to adequately characterize the cell suspensions. Therefore a combination of both methods plus the determination of the mitotic index were employed to follow the cell cycle kinetics of populations which were submitted to perturbing conditions. © 1989, Gustav Fischer Verlag, Stuttgart. All rights reserved.

Author-supplied keywords

  • 4′-,6-diamidino-2-phenylindole
  • BrdUrd
  • DAPI
  • FITC
  • MI
  • Nicotiana tabacum
  • PBS
  • PBT
  • bromodeoxyuridine
  • cell cycle analysis
  • cell suspension culture
  • flow cytometry
  • fluorescein isothiocyanate
  • mitotic index
  • monoclonal antibody to bromodeoxyuridine
  • phosphate buffered saline
  • phosphate buffered saline +0.5% Triton X-100
  • synchronization

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Authors

  • Martin Pfosser

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