Focal plane tuning in wide-field-of-view microscope with Talbot pattern illumination.

We have developed a focal plane tuning technique for use in focus-grid-based wide-field-of-view microscopy (WFM). In WFM, the incidence of a collimated beam on a mask with a two-dimensional grid of aperture produced the Talbot images of the aperture grid. The Talbot pattern functioned as a focus grid and was used to illuminate the sample. By scanning the sample across the focus grid and collecting the transmission, we can generate a microscopy image of the sample. By tuning the wavelength of the laser, we can tune the focal plane of the WFM and acquire images of different depth into the sample. Images of a green algae microscope slide were acquired at different focal planes for demonstration.