Counting pollen grains using readily available, free image processing and analysis software
Counting pollen grains using readily available, free image processing and analysis software
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DOI：
10.1093/aob/mcp186
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发表时间：
2009-10-01
期刊：
影响因子：
4.2
通讯作者：
Yang, Suann
中科院分区：
文献类型：
--
作者：
Costa, Clayton M.;Yang, Suann
虽然存在许多方法来定量样品中的花粉粒数，但很少有标准的方法是用户友好的，廉价的和可靠的。本文描述了一种利用现成的免费图像处理和分析软件计算花粉的新方法。采集了两种菊科植物Carduus acanthoides和C. nutans的花药花粉，在载玻片上进行光照，并在立体显微镜下进行数码摄影。使用ImageJ （NIH）对这些数字图像进行处理以去除噪声并锐化单个花粉粒，然后进行分析以获得图像中存在的可靠的花粉粒总数。开发了一个宏来同时分析多个图像。为了评估ImageJ分析的花粉计数的准确性和一致性，将计数与人眼计数进行了比较。图像分析在每张图像60秒或更短的时间内产生花粉计数，比人眼计数（5-68分钟）快得多。此外，ImageJ程序产生的计数与肉眼获得的计数相似。由于计数参数可调，该图像分析方案可用于许多其他植物物种。因此，该方法为从数字图像中计数花粉提供了一种快速、廉价和可靠的解决方案，不仅减少了错误的机会，而且大大降低了劳动力需求。
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.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.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.