Device and method for spectral imaging

A spectral imaging device comprising:

• an imager configured to capture image frames;
• a scanning stage configured to establish relative motion between the imager and a sample in a scanning direction;
• and
• an optical system configured to control a light characteristic of a light beam constituting an image of the sample to the imager, the optical system comprising:
• a light varying element configured to receive the light beam and provide an output light beam having a temporal varying light characteristic;
• and
• a set of redirecting optical elements configured to redirect light rays directed from the sample to form the light beam, and to focus the output light beam onto the imager;
• and
• a controller configured to control the scanning stage, the light varying element and the imager to capture a plurality of image frames with movement of the scanning stage in a scanning direction and with changes in the light characteristics that span a defined spectrum, wherein the plurality of image frames is captured with an overlap including a defined shift that is greater than 1 pixel along the scanning direction between consecutive image frames in the plurality of image frames.

A method of analyzing a spectral image of a sample having cells stained by a stain, the method comprising:

• identifying in the spectral image a plurality of nuclei, and extracting, for each nucleus, a spectrum characterizing an optical transmission and absorbance of the nucleus within a wavelength range corresponding to the stain, thereby providing a plurality of spectra, one spectrum for each nucleus of at least a portion of the nuclei;
• comparing shapes of the spectra thereamongst;
• identifying each nucleus of the portion of the nuclei, as belonging to one of at least a first population of nuclei and a second population of nuclei based on the comparing;
• and
• marking the nucleus according to the identification.