Methods for cellularly addressable nucleic acid sequencing

A system for sequencing nucleic acids, comprising:

• an optical system comprising:
• an objective lens having a field-of-view (FOV), wherein the objective lens provides less than 0.15 waves of aberration across at least 80% of the FOV at a fluorescent emission wavelength and a Strehl ratio of at least 0.8 across at least 80% of the FOV;
• an excitation energy source configured to illuminate one or more biological samples immobilized on a flow cell;
• at least one optical sensor that senses fluorescent emission signals of the one or more biological samples, the one or more biological samples comprising one or more cells, and the one or more cells comprising a plurality of cellular nucleic acid molecules at locations within the one or more cells of the biological samples, wherein the fluorescent emission signals comprise:
• fluorescent intensities and corresponding spatial locations thereof on the flow cell;
• and
• a rolling circle amplification reagent and a plurality of concatemer molecules that are generated from the plurality of cellular nucleic acids, wherein the rolling circle amplification reagent comprises (i) a plurality of strand-displacing polymerases and (ii) a plurality of nucleotides.