Systems and methods for hyperspectral imaging are described. A hyperspectral imaging system (300), comprising: a sample holder (370) configured to hold a sample; a light source (3010) configured to emit excitation light (402) having one or more wavelengths; a two-dimensional imaging device (380); and an optical system comprising: a first spatial light modulator (320a); a first dispersive element (340a); and a second dispersive element (340b); wherein the optical system is configured to (i) structure the excitation light (402), using the first spatial light modulator (320a), into a predetermined two-dimensional excitation pattern at a conjugate plane of a focal plane in the sample; (ii) spectrally disperse, using the first dispersive element (340a), the excitation light (404); (iii) use excitation pattern (406) to illuminate the sample illuminate the sample in an excitation pattern (406) with the one or more wavelengths dispersed along a first lateral direction within the focal plane; (iv) collect, from the sample, emission light (408); (v) spectrally disperse, using the second dispersive element (340b), the collected emission light (408); and (vi) provide the collected emission light (410) in-focus to the imaging device (380) such that the spectrally dispersed emission light (410) is spectrally dispersed along a second lateral direction.