Devices and compositions for use in luminescent imaging of multiple sites within a pixel, and methods of using the same

A device for use in luminescent imaging, the device comprising:

• an imaging pixel;
• a first nanowell corresponding to the imaging pixel, and
• a second nanowell corresponding to the imaging pixel, the second nanowell being spatially displaced from the first nanowell;
• a first luminophore disposed within or over the first nanowell;
• a second luminophore disposed within or over the second nanowell;
• at least one radiation source configured to generate first photons having a first characteristic, and configured to generate second photons having a second characteristic, the second characteristic being different than the first characteristic;
• and
• a controller coupled to the at least one radiation source and configured to cause the at least one radiation source to illuminate the first and second features nanowells with the first photons at a first time and to illuminate the first and second features nanowells with the second photons at a second time which is different from the first time,
• wherein the first photons interfere with one another in such a manner as to selectively illuminate the first nanowell with a first interference pattern, the first interference pattern having a field strength that is more intense at the first nanowell than at the second nanowell and thus selectively excites the first luminophore to a greater extent than the second luminophore at the first time, and
• wherein the second photons interfere with one another in such a manner as to selectively illuminate the second nanowell with a second interference pattern, the second interference pattern having a field strength that is more intense at the second nanowell than at the first nanowell and thus excites the second luminophore to a greater extent than the first luminophore at the second time, the second interference pattern being different than the first interference pattern;
• and
• wherein the imaging pixel is configured to receive luminescence emitted by the first luminophore of the first nanowell responsive to the selective illumination of the first nanowell by the first interference pattern at the first time, and is configured to receive luminescence emitted by the second luminophore of the second nanowell responsive to selective illumination of the second nanowell by the second interference pattern at the second time.