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In contrast to brightfield microscopy, darkfield microscopy allows observation of living specimens. Consistent with the results of TEM observation, the distribution of the scattering intensity of each spot obtained with TIRDFM showed a single peak with a small population with a higher intensity . The scattering intensity depends strongly on the radius r of the scattering object and is proportional to r6 . The lower variation in the scattering intensity further validates that most scattering spots were derived from single viral particles. In primary syphilis, the chancre teems with treponemes that can be seen with darkfield microscopy. Sample material needs to be spread thinly, dense preparations can grossly affect the contrast and accuracy of the dark field’s image. Components of Bright field microscope and difference in illumination with darkfield microscope. Definitive identification of T pallidum depends on visualizing not only its typical morphology but also its typical motility

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