Single-Molecule Fluorescence Microscopy

In recent years, many different optical methods have been developed to image individual molecules with sub-wavelength resolution. In our experiments we have mostly relied on a derivative of FIONA (Fluorescence Imaging with One Nanometer Accuracy [1]) and STORM (Stochastic Optical Reconstruction Microscopy [2]), where molecules arrive such as to never lie next to each other during the photo-cycle lifetime. The point spread function of every molecule can then be fitted by a smooth curve whose center may be determined with better than 10 nm accuracy.

We have used Laser-Induced Epi-Fluorescence microscopy on a dilute molecular sample to detect a de Broglie interferogram with single-molecule resolution in real time [3]. 

  • A. Yildiz, P.R. Selvin,
    Fluorescence imaging with one nanometer accuracy: application to molecular motors,
    Acc. Chem. Res. 38,  574-582 (2005).
  • Rust, M. J., Bates, M. and Zhuang, X. 
    Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). 
    Nature Methods 3: 793-796 (2006).
  •  T. Juffmann, A. Milic, M. Müllneritsch, P. Asenbaum, A. Tsukernik, J. Tüxen, M. Mayor, O. Cheshnovsky, M. Arndt,
    Real-time single-molecule imaging of quantum interference
    Nature Nanotechn. 7,  297 - 300 (2012).