Tunable depth illumination with multi-angle TIRF and epi-fluorescence implemented on selective regions in the field-of-view

Despite the great depth sectioning performance of total-internal-reflection-fluorescence (TIRF) microscopy, it is difficult to measure signals distributed in various depths using conventional TIRF. We propose a depth-tunable imaging scheme that can simultaneously implement TIRF with multiple illumination depths and epi-fluorescence illumination in user chosen regions.