A method of imaging described herein comprises (a) disposing ultrasound-switchable fluorophores within an environment; (b) exposing the environment to ultrasound to create a first activation region within the environment; (c) disposing the fluorophores within the first activation region to switch the fluorophores from an off state to an on state; (d) irradiating the environment to excite the fluorophores within the first activation region; (e) detecting photoluminescence from the excited fluorophores at a first optical spot on an exterior surface of the environment; (f) subsequently creating a second activation region within the environment; (g) switching fluorophores within the second activation region to an on state; (h) exciting the fluorophores in the on state within the second activation region; and (i) detecting photoluminescence from the excited fluorophores within the second activation region at a second optical spot on the exterior surface, wherein the first and second optical spots are optically resolvable.