Journal Article

Counting pollen grains using readily available, free image processing and analysis software

Clayton M. Costa and Suann Yang

in Annals of Botany

Published on behalf of The Annals of Botany Company

Volume 104, issue 5, pages 1005-1010
Published in print October 2009 | ISSN: 0305-7364
Published online July 2009 | e-ISSN: 1095-8290 | DOI: https://dx.doi.org/10.1093/aob/mcp186
Counting pollen grains using readily available, free image processing and analysis software

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  • Ecology and Conservation
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Background and Aims

Although many methods exist for quantifying the number of pollen grains in a sample, there are few standard methods that are user-friendly, inexpensive and reliable. The present contribution describes a new method of counting pollen using readily available, free image processing and analysis software.

Methods

Pollen was collected from anthers of two species, Carduus acanthoides and C. nutans (Asteraceae), then illuminated on slides and digitally photographed through a stereomicroscope. Using ImageJ (NIH), these digital images were processed to remove noise and sharpen individual pollen grains, then analysed to obtain a reliable total count of the number of grains present in the image. A macro was developed to analyse multiple images together. To assess the accuracy and consistency of pollen counting by ImageJ analysis, counts were compared with those made by the human eye.

Key Results and Conclusions

Image analysis produced pollen counts in 60 s or less per image, considerably faster than counting with the human eye (5–68 min). In addition, counts produced with the ImageJ procedure were similar to those obtained by eye. Because count parameters are adjustable, this image analysis protocol may be used for many other plant species. Thus, the method provides a quick, inexpensive and reliable solution to counting pollen from digital images, not only reducing the chance of error but also substantially lowering labour requirements.

Keywords: Carduus acanthoides; Carduus nutans; ImageJ; image processing; pollen grain count

Journal Article.  3902 words.  Illustrated.

Subjects: Ecology and Conservation ; Evolutionary Biology ; Plant Sciences and Forestry

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