Digitally enhanced microscopy for multiplexed histology

An imaging system for imaging a sample on a microscope slide, comprising:

• at least one processor;
• and
• memory storing program instructions that, when executed by the at least one processor, cause the imaging system to:
• cause a plurality of light sources of an optical system to output different light wavebands at different times to sequentially illuminate the sample, wherein the plurality of light sources includes one or more of an ultraviolet light source, a near infrared light source, or an infrared light source, wherein the optical system includes a plurality of filters configured for individual color channels associated with respective one of the light sources;
• obtain a first image of the sample exposed to a first light waveband from a first light source of the plurality of light sources and using a first filter of the plurality of filters, wherein the first light waveband is configured to interact with a first target of the sample, wherein the first target is a first chromogen or a first fluorophore,
• obtain a second image of the sample exposed to a second light waveband from a second light source of the plurality of light sources and using a second filter of the plurality of filters, wherein the second light waveband is configured to interact with a second target of the sample, wherein the second target includes a second chromogen or a second fluorophore, the second light waveband being different from the first light waveband,
• generate an enhanced color composite image of the sample with a false color for at least one of the first or second targets, such that color contrast between the first and second targets in the enhanced color composite image is increased compared to natural color contrast between the first and second targets in the first and second images, and
• output the enhanced color composite image.

A method for producing a multicolor image of a sample, the method comprising:

• capturing images of the sample at a rate greater than or equal to two images per second, wherein the captured images include
• a first image of the sample exposed to a first light waveband from a first light source,
• wherein the first light waveband is an ultraviolet waveband, a near infrared waveband, or an infrared waveband, wherein the sample includes a first feature stained with a first chromogen or a first fluorophore configured to interact with the first light waveband;
• a second image of the sample exposed to a second light waveband from a second light source,
• wherein the second light waveband is different from the first light waveband, wherein the sample includes a second feature stained with a second chromogen or second fluorophore configured to interact the second light waveband;
• producing a digitally-enhanced multicolor image of the sample from the first and second images by converting at least one of the first or second images into a false color image in which color contrast between the first and second features in the digitally-enhanced multicolor image is increased compared to natural color contrast between the first and second features in the first and second images.