We harness dual-faceted optical manipulation—manipulation by light and of light—in spectroscopy and microscopy to reveal biological structures and functions with exceptional sensitivity, resolution, and speed. Enhanced optical microscopy with rotation and machine learning enables volumetric imaging and precise organism classification. An adhesion frequency assay profiles cell-cell interactions, while chiroptical spectroscopy offers label-free, ultrasensitive enantiodiscrimination of chiral molecules. These innovations expand optical measurement capabilities, advancing life sciences, space exploration, pharmaceutical quality control, and disease diagnosis.