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Pollen Grain Counting Using a Cell Counter

  • Hiroyuki Kakui
  • Misako Yamazaki
  • Naoto-Benjamin Hamaya
  • Kentaro K. ShimizuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2160)

Abstract

The number of pollen grains is a critical part of the reproductive strategies in plants and varies greatly between and within species. In agriculture, pollen viability is important for crop breeding. It is a laborious work to count pollen tubes using a counting chamber under a microscope. Here, we present a method of counting the number of pollen grains using a cell counter. In this method, the counting step is shortened to 3 min per flower, which, in our setting, is more than five times faster than the counting chamber method. This technique is applicable to species with a lower and higher number of pollen grains, as it can count particles in a wide range, from 0 to 20,000 particles, in one measurement. The cell counter also estimates the size of the particles together with the number. Because aborted pollen shows abnormal membrane characteristics and/or a distorted or smaller shape, a cell counter can quantify the number of normal and aborted pollen separately. We explain how to count the number of pollen grains and measure pollen size in Arabidopsis thaliana, Arabidopsis kamchatica, and wheat (Triticum aestivum).

Key words

Pollen number Pollen size Pollen viability Cell counter (CASY cell counter) Arabidopsis thaliana Arabidopsis kamchatica Wheat (Triticum aestivum

Notes

Acknowledgements

This study was supported by the Swiss National Science Foundation 31003A_182318, a JST CREST Grant Number JPMJCR16O3 and MEXT KAKENHI Grant Number 16H06469 to KKS, and a MEXT KAKENHI Grant Number 19K05976 to H.K. We thank Beat Keller for plant material and Reiko Akiyama and Toshiaki Tameshige for their valuable suggestions.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Hiroyuki Kakui
    • 1
    • 2
  • Misako Yamazaki
    • 1
  • Naoto-Benjamin Hamaya
    • 1
  • Kentaro K. Shimizu
    • 1
    • 2
    Email author
  1. 1.Department of Evolutionary Biology and Environmental StudiesUniversity of ZurichZurichSwitzerland
  2. 2.Kihara Institute of Biological ResearchYokohama City UniversityYokohamaJapan

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