Spatially-multiplexed fluorescence imaging

A method of performing fluorescence imaging of a sample using a fluorescence imaging device, the sample having a first sample portion supported by a first sample well of a first sample well region and a second sample portion supported by a second sample well of a second sample well region, and the method comprising:

• (a) directing an imaging aperture of the fluorescence imaging device toward the first sample well region;
• (b) after (a), capturing, using the imaging aperture, a first image of the first sample well region;
• (c) after (b), redirecting the imaging aperture toward the second sample well region;
• and
• (d) after (c), capturing, using the imaging aperture, a second image of the second sample well region.

A fluorescence imaging system, comprising:

• a first sample well region comprising a first sample well configured to support a first portion of a sample;
• a second sample well region comprising a second sample well configured to support a second portion of the sample;
• and
• a fluorescence imaging device configured to:
• select a sample well region from among the first sample well region and the second sample well region;
• and
• capture an image of the sample well region.

A method of polypeptide sequencing, the method comprising:

• (a) capturing, by a fluorescence imaging device, a fluorescence image of a first sample well;
• and
• (b) in a second sample well, performing a controlled cleavage of a terminal amino acid of a polypeptide immobilized to a surface of the second sample well, the controlled cleavage comprising catalyzing removal of the terminal amino acid of the polypeptide,
• wherein (a) is performed during at least a portion of (b).

A fluorescence imaging device, comprising:

• a first fluorescence camera configured to capture light contained within a first optical band;
• a second fluorescence camera configured to capture light contained within a second optical band;
• and
• optical components configured to:
• receive fluorescent light emitted by a sample, the fluorescent light comprising a first portion having content contained within the first optical band and a second portion having content contained within the second optical band;
• provide the first portion of the fluorescent light to the first fluorescence camera;
• and
• provide the second portion of the fluorescent light to the second fluorescence camera.

A fluorescence imaging device, comprising:

• a first fluorescence camera;
• a second fluorescence camera;
• and
• optical components configured to receive fluorescent light emitted by a sample and divide the fluorescent light between a first optical path that includes the first fluorescence camera and a second optical path that includes the second fluorescence camera.

A fluorescence imaging system comprising:

• a plurality of sample well regions;
• a controller configured to:
• select a sample well region of the plurality of sample well regions;
• and
• dispense a sample, reagent and/or buffer into the sample well region;
• and
• a fluorescence imaging device configured to capture fluorescent light emitted from the sample well region following dispensation of the sample, reagent, and/or buffer.

A fluorescence imaging and/or processing system, comprising:

• processing circuitry configured to:
• obtain a first image comprising a first plurality of pixel values indicating intensity of fluorescent light received at respective pixels of a fluorescence imaging device;
• and
• transform the first image into a second image comprising a second plurality of pixel values indicating intensity of fluorescent light received from respective sample wells of a sample well member.

A fluorescence imaging and/or processing system, comprising:

• processing circuitry configured to:
• obtain a fluorescence image captured by a fluorescence imaging device indicating fluorescent light emitted from a sample well member;
• determine, based on the fluorescence image, a position of the fluorescence imaging device with respect to the sample well member;
• and
• output a signal indicating an extent of alignment and/or misalignment between the fluorescence imaging device and the sample well member.