Multispectral sample imaging

A method, comprising:

• exposing a plurality of non-endogenous spectral contributors in a biological sample to illumination light and measuring light emission from the sample to obtain N sample images, wherein each sample image corresponds to a different combination of a wavelength band of the illumination light and one or more wavelength bands of the light emission, wherein multiple wavelength bands of the light emission from the plurality of non-endogenous spectral contributors define a wavelength range, and wherein N>1;
• and
• exposing the sample to illumination light in a background excitation band and measuring light emission from the sample in a background spectral band to obtain a background image of the sample, wherein the background spectral band comprises a distribution of wavelengths having a full width at half maximum (FWHM) spectral width Δλ and a center wavelength λc within the wavelength range, and wherein the wavelengths within the background spectral band correspond to wavelengths within a range from λc−Δλ/2 to λc+Δλ/2,
• wherein for each of the plurality of non-endogenous spectral contributors in the sample exposed to the illumination light in the background excitation band, a spectral emission intensity at each wavelength within the background spectral band is 10% or less of a maximum measured spectral emission intensity of the non-endogenous spectral contributor following excitation of the non-endogenous spectral contributor in each of the wavelength bands of the illumination light and the background excitation band.