Semi-automated measurement of motility of human subgingival microflora by image analysis

The purpose of this investigation was to quantitatively estimate bacterial motility by image analysis, and to apply this method for the measurement of motility of human subgingival microflora. We developed a semi-automated method for the quantification of bacterial motility using video microscopy, digitization and image processing. Moving images of both authentic bacterial samples and clinical samples were recorded using a phase contrast microscope with a high speed (1/100 s) shutter camera. The motility was evaluated by measuring the total number of pixels remaining after the subtraction of 2 serial video images. The total number of pixels was significantly correlated with both the sum of the velocity of each bacterial cell and the number of motile bacteria on the same original images. Motility of subgingival microflora from 140 clinical samples tested was measured at 0 pixels to 3600 pixels, whereas the effect of Brownian movement was less than 150 pixels. The motility of subgingival microflora estimated with this image analysis system did not differ much from objective judgments by the naked eyes of experts. These results suggest that a semi-automated image analysis system may be useful in the evaluation of the motility of human subgingival microflora.