Digitally enhanced microscopy for multiplexed histology

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

• a slide imager configured to generate an enhanced composite image of the sample, the slide imager including:
• an energy emitter including a first light source and a second light source;
• one or more image capture devices;
• and
• a controller communicatively coupled to the energy emitter and the one or more image capture devices, the controller including at least one processor and memory storing program instructions that, when executed by the at least one processor, cause the slide imager to perform a process including:
• capturing, using the one or more image capture devices,
• a first monochrome image of the sample exposed to a first light waveband from the first light source, wherein the first light waveband interacts with a first target of the sample, and
• a second monochrome image of the sample exposed to a second light waveband from the second light source, wherein the second light waveband interacts with a second target of the sample, wherein the second light waveband is different from the first light waveband,
• combining the first target illustrated in a first false color and the second target illustrated in a second false color different than the first false color to generate the enhanced composite image of the sample, and
• identifying, using a trained classifier, one or more features of interest in the enhanced composite image.

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

• an energy emitter including a light source;
• one or more image capture devices;
• and
• a controller communicatively coupled to the energy emitter and the one or more image capture devices, the controller including at least one processor and memory storing instructions that, when executed by the at least one processor, cause the imaging system to perform a process including:
• determining, using the at least one processor, a first light waveband for interacting with a first target of the sample and a second light waveband for interacting with a second target of the sample,
• performing an illumination protocol via the light source to sequentially illuminate the sample with the first light waveband and the second light waveband,
• capturing, using the one or more image capture devices, a first monochrome image and a second monochrome image of the sample, wherein the first monochrome image is captured while the sample is illuminated with the first light waveband and the second monochrome image is captured while the sample is illuminated with the second light waveband, and
• generating an enhanced image of the sample with the first target illustrated in a first false color and the second target illustrated in a second false color different than the first false color.

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

• an energy emitter including a first light source and a second light source;
• one or more image capture devices;
• and
• a controller communicatively coupled to the energy emitter and the one or more image capture devices, the controller including at least one processor and memory storing a plurality of false color protocols and instructions that, when executed by the at least one processor, cause the imaging system to perform a process including:
• capturing, using the one or more image capture devices,
• a first monochrome image of the sample exposed to a first light waveband from the first light source, wherein the first light waveband interacts with a first feature of the sample, and
• a second monochrome image of the sample exposed to a second light waveband from the second light source, wherein the second light waveband interacts with a second feature of the sample, wherein the second light waveband is different from the first light waveband,
• selecting at least one false color protocol representing a multi-feature staining protocol for visually identifying multiple features of the sample;
• assigning a first false color for the first feature according to the at least one false color protocol,
• assigning a second false color for the second feature according to the at least one false color protocol, wherein the second false color is different from the first false color according to the at least one false color protocol,
• combining the first feature and the second feature illustrated in the first and second false colors to generate a labelled false color image of the sample, wherein the labelled false color image of the sample includes the first false color first feature and the second false color second feature, and
• outputting an enhanced composite image.