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Dep. of Horticulture, Univ. of Arkansas, 316 Plant Sciences Bldg., Fayetteville, AR 72701
* Corresponding author (karcher{at}uark.edu)
| ABSTRACT |
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| INTRODUCTION |
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Under normal use of SigmaScan Pro, each image must be processed manually, which may take several minutes and limit large-scale use of the techniques. However, SigmaScan Pro is enabled with a macro language so that users may automate software processes. The development of a user-friendly, easily attainable macro to batch analyze large numbers of turf images should broaden the use of digital image analysis techniques and result in more objective evaluations of turfgrass trials. The objective of the following work was to develop such a macro.
| MATERIALS AND METHODS |
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Before executing the macro, the images to be analyzed must all share the same file extension and be named so that they share a root name and each image contains a unique, consecutive number at the end of the file name (example: Plot01.jpg, Plot02.jpg, Plot03.jpg...). Most digital image archiving software packages that are sold with digital cameras are capable of naming or renaming image files similar to the format described above. In addition, there are many freeware file renaming software packages currently available for download (Nonags.com, 2004). The macro will function correctly with or without leading zeros at the end of the file name and with any image file extension that SigmaScan Pro recognizes (.jpg,.bmp,.tif, etc...).
The macro is executed by selecting "File > Open > Macro" from within SimgaScan Pro. Upon selecting the "Turf Analysis" macro file and then pressing the "F8" key to begin macro execution, a dialog box named "Turf Analysis Settings" is opened (Fig. 1) to customize the macro settings for a specific analysis of a batch of images.
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Color Analysis
An analysis of turf color will be performed on the images if the "Perform color analysis" (Fig. 1) checkbox is selected. Either entire images may be used to determine average color or a color threshold may be used to determine average color from selected pixels. In the example analysis settings, color analysis of the brown patch images will be performed using threshold settings so that only green pixels (representing nondiseased turf) are used to calculate average color. The default macro settings perform a color analysis using the entire image to calculate average color.
Cover Analysis
An analysis of percent turf cover will be performed on the images if the "Perform cover analysis" (Fig. 1) checkbox is selected. Three options are available for determining the total number of pixels in each image, which is subsequently used to calculate percent cover: The first option is to separately measure the total pixel number for each image, which is the slowest option, but is necessary if total pixel numbers vary among the images within a batch. This would be the case if pictures within a batch were taken with different resolution settings or if images were cropped before analysis. The second option is to only measure the total pixel number for the first image and use that value as the total pixel number for all subsequent images, which is appropriate if all images within an analysis batch were taken with identical camera settings, not cropped, and thus share the same number of total pixels. The third and fastest option, which may be used if all images share the same number of total pixels, is to manually enter the shared total pixel number in the text box provided. The default macro settings analyze images for percent cover and only measure the first image to determine total pixel numbers.
Threshold Settings
The "Threshold Settings" box (Fig. 1) is used to adjust the hue and saturation levels that are used to select pixels representing green turf for the cover and color analyses. Before executing the macro, it is important to open a few representative images in SigmaScan Pro and select "Image > Threshold > Color Threshold" to determine the precise hue and saturation levels that will select the pixels of interest within the images. The appropriate threshold settings will vary depending on factors such as turf species and variety, management practices, light conditions present when images were collected, and camera model and settings. The default settings are hue levels from 30 to 100 and saturation levels from 0 to 100.
If the "Save output as Excel file in image folder" box (Fig. 1) is checked, the macro output will be automatically saved to the folder containing the batch of images to be analyzed. The default macro setting is to save the output as a Microsoft Excel file. Clicking the "OK" button will begin the image analysis process.
| RESULTS AND DISCUSSION |
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The example analysis accomplished with the "Turf Analysis" macro, which evaluated 489 images for percent turf cover and turf color, required less than one minute of manual execution. At an average rate of three minutes per image to manually evaluate percent cover and color in SigmaScan Pro, the "Turf Analysis" macro saved more than 24 h of manual, tedious labor. Although the macro can be executed in less than one minute, the analysis will complete at a rate from <5 s to 1 min. per image, depending on the resolution of the images and the computer processing speed.
Although the "Turf Analysis" macro was developed for the analysis of turf images, its utility may be extended to analyze virtually any plant species. Through collaboration efforts with colleagues from the departments of Horticulture and Crop, Soil, and Environmental Sciences at the University of Arkansas, Fayetteville, the macro has been tested successfully in quantifying the colors of poinsettia (Euphorbia pulcherrima Willd.) and petunia (Petunia spp. Juss.) flowers, spinach leaves (Spinacia oleracea L.) and quantifying soybean [Glycine max (L.) Merr.] canopy coverage.
Received for publication September 21, 2004.
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