Saturated absorption competition microscopy

Abstract: We introduce the concept of saturated absorption competition (SAC) microscopy as a means of providing sub-diffraction spatial resolution in fluorescence imaging. Unlike the post-competition process between stimulated and spontaneous emission that is used in stimulated emission depletion (STED) microscopy, SAC microscopy breaks the diffraction limit by emphasizing a pre-competition process that occurs in the fluorescence absorption stage in a manner that shares similarities with ground-state depletion (GSD) microscopy. Moreover, unlike both STED and GSD microscopy, SAC microscopy offers a reduction in complexity and cost by utilizing only a single continuous-wave laser diode and an illumination intensity that is ~ 20x smaller than that used in STED. Our approach can be physically implemented in a confocal microscope by dividing the input laser source into a time-modulated primary excitation beam and a doughnut-shaped saturation beam, and subsequently employing a homodyne detection scheme to select the modulated fluorescence signal. Herein, we provide both a physico-chemical model of SAC and experimentally demonstrate by way of a proof-of-concept experiment a transverse spatial resolution of ~lambda/6.