We harness dual-faceted optical manipulation—using light to control matter and matter to modulate light—to advance spectroscopy and microscopy with high sensitivity, resolution, and speed. Our microscopy platform integrates optical rotation and machine learning for volumetric imaging and precise organism classification. We develop adhesion frequency assays to quantify dynamic cell–cell interactions and enhance chiroptical spectroscopy for label-free, ultrasensitive enantiodiscrimination. These innovations drive progress in biomedical research, pharmaceutical analysis, space biology, and clinical diagnostics.