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Superresolution Imaging with Single-Antibody Labeling.

Hirushi GunasekaraThilini PereraJesse AndersonBadeia SaedNeal RamseierNeama KeshtaYing Samuel Hu
Published in: Bioconjugate chemistry (2023)
We present a versatile single-molecule localization microscopy technique utilizing time-lapse imaging of single-antibody labeling. By performing single-molecule imaging in the subminute time scale and tuning the antibody concentration to create sparse single-molecule binding, we captured antibody labeling of subcellular targets to generate superresolution images. Single-antibody labeling enabled dual-target superresolution imaging using dye-conjugated monoclonal and polyclonal antibodies. We further demonstrate a dual-color strategy to increase the sample labeling density. Single-antibody labeling paves a new way to evaluate antibody binding for superresolution imaging in the native cellular environment.
Keyphrases
  • single molecule
  • high resolution
  • atomic force microscopy
  • living cells
  • machine learning
  • deep learning
  • convolutional neural network
  • transcription factor
  • dna binding
  • single cell