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Counting Colonies or Plaques Directly from Images of Agar Plates

contrast between plaques and the bacterial lawn for precise analysis. In this case, an image with improved contrast can be acquired using overhead white-light illumination while the agar dish is resting on top of a piece of black, matte plastic. This method can also be used for acquiring images of small, abundant, white bacterial colonies on agar plates to increase contrast and enhance counting precision. Alternatively, plaque plates can be moved closer to the camera lens manually while using white-light-box transillumination. During all acquisition methods, the magnification should be adjusted so that the edges of the petri dish are fully visible and centered in the image. This adjustment will aid the automatic determination of the counting region, which is based on detecting the edge of the agar dish.

Defining Image Type

A critical factor in obtaining an accurate count is correctly defining the image type prior to analysis. When colonies or plaques appear lighter than the background of the plate, the image should be defined as white on black. Conversely, when colonies or plaques appear darker than the background of the plate, the image should be defined as black on white. If the image type is improperly defined, the count will not be accurate, and colonies or plaques that are counted will not correspond to those that appear on the image. When the Colony/Plaque Counting function is first enabled, an Image Type Selection dialog box is displayed, which prompts the user to specify image type. Before entering the Colony/Plaque Counting function, the image type can also be changed using the toolbar icon or the Analyze menu.

Conclusions

In addition to molecular weight and basic densitometry functions, the analysis portion of Eagle Sight software version 3.1 now includes a colony- and plaque-counting function. This function allows researchers to count colonies and plaques directly fro
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