Systems and methods for 4-D hyperspectral imaging

A hyperspectral imaging system comprising:

• a sample holder configured to hold a sample;
• a light source configured to emit excitation light having one or more wavelengths;
• a two-dimensional (2-D) imaging array;
• and
• an optical system comprising:
• a first spectral separator comprising a first dispersive element or a first diffractive element;
• and
• a second spectral separator comprising a second dispersive element or a second diffractive element;
• wherein the optical system is configured to (i) structure the excitation light, using the first spectral separator, into a spectrally dispersed two-dimensional excitation pattern at a focal plane in the sample or a conjugate plane of the focal plane;
• (ii) use the excitation pattern to illuminate the sample with the one or more wavelengths dispersed along a first direction within the focal plane;
• (iii) collect, from the sample, emission light;
• (iv) spectrally disperse, using the second spectral separator, the collected emission light;
• and (v) provide the collected emission light in-focus to the imaging array such that the collected emission light is spectrally dispersed along a second direction.