Systems and methods for use in tri-spot point spread function imaging are provided that include a phase mask. The phase mask includes three partitions that each include a subset of the total area that is asymmetrical to other partitions. Each partition includes a phase delay ramp aligned along a phase delay axis, and includes a gradient of phase delays. Each phase delay axis is oriented in a different direction with respect to each other. Included is a source that outputs an excitation beam into a sample containing at least one light emitter that emits a radiation pattern when illuminated. Included is at least one sensor arranged to capture at least one image of the radiation pattern and a phase mask positioned between the at least one emitter and the at least one sensor. The phase mask is configured to produce a tri-spot point spread function.