A detailed instruction is described on how to build a highly inclined swept tile (HIST) microscope and its usage for single-molecule imaging.
Prepare optical table, mount core components, and establish foundational alignment before optical path configuration.
Install filters, dichroics, and detectors to collect single-molecule fluorescence signals from the specimen.
Align laser excitation lines and integrate cylindrical lenses to create the highly inclined illumination geometry.
Validate multi-region imaging by acquiring sequential tile scans to confirm coverage and signal quality.
Execute HIST imaging protocol to generate extended field-of-view single-molecule data with reduced background.
Demonstrate imaging performance on labeled DNA and RNA to confirm visualization of single molecules.
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