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Significance The precision of localization-based superresolution microscopy techniques fundamentally relies on the point-spread function of the optical system and the number of photons one can collect. Here, we report that by using a high-numerical-aperture objective lens and a custom cryogenic stage, we are able to increase photon yield by 2–3-fold over room temperature, thereby achieving more precise superresolution reconstructions of complex subcellular structures.

High-numerical-aperture cryogenic light microscopy for increased precision of superresolution reconstructions