Classification and counting of fluorescent pollen using an image analysis system
Classification and counting of fluorescent pollen using an image analysis system
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DOI：
10.1080/714028122
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发表时间：
2001-01-01
影响因子：
1.6
通讯作者：
Eduardo, P
中科院分区：
文献类型：
--
作者：
Aronne, G;Cavuoto, D;Eduardo, P
由于花粉粒的阳性荧光与其萌发能力之间的高度相关性，因此通常用荧光反应(FCR)来评估花粉的生活力。这种方法的优点之一是它的简单性。然而，一项测试FCR分析重复性的实验表明，结果受到主观性的影响。分析员之间几乎没有一致性，同一分析员对在不同时间检查的同一花粉样本图像的评估可能不同。这些问题被计算机图像分析系统解决了，该系统提供了一种对正负荧光花粉进行分类并对每一类中的颗粒进行自动计数的方法。计算机图像分析系统不改变FCR检测的生物化学，但避免了由于分析员的主观性造成的一些实验误差。FCR后花粉的显微镜图像被数字化，然后通过专门设计的软件“植物测量仪”进行分析。该软件删除图像的暗背景以分离颗粒，然后计算正负荧光花粉颗粒。对软件输出进行了实验验证，结果可靠。此外，该软件对用户友好，分析师只需很少的培训就能获得可靠的结果。
Pollen viability is commonly assessed by fluorochromatic reaction (FCR) because of the high correlation between positive fluorescence of the pollen grains and their ability to germinate. One of the advantages of this method is its simplicity. An experiment to test FCR analysis for reproducibility, however, showed that results are affected by subjectivity. There is little consistency between analysts, and assessment by the same analyst may differ for the same pollen sample image examined at different times. These problems were solved by a computerized image analysis system that provides a method for classifying positive and negative fluorescent pollen and automatic counting of the grains in each class. The computerized image analysis system does not change the biochemistry of the FCR test, but avoids some experimental errors owing to the subjectivity of the analyst. Microscope images of the pollen after FCR were digitized and later analyzed by specially designed software, "Plant Meter." This software deletes the dark background of the image to isolate the grains, and subsequently counts positive and negative fluorescent pollen grains. An experiment was carried out to validate software output and it showed reliable results. Moreover, the software is user friendly and very little training is necessary for analysts to achieve reliable results.