Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes

1. Sang-Hee Shim^a,¹,
2. Chenglong Xia^b,¹,
3. Guisheng Zhong^a,^c,
4. Hazen P. Babcock^a,^d,
5. Joshua C. Vaughan^a,^c,
6. Bo Huang^a,²,
7. Xun Wang^b,
8. Cheng Xu^b,
9. Guo-Qiang Bi^b,³, and
10. Xiaowei Zhuang^a,^c,^d,^e,³

+ Author Affiliations

1. ^aDepartment of Chemistry and Chemical Biology,
2. ^cHoward Hughes Medical Institute,
3. ^dCenter for Brain Science, and
4. ^eDepartment of Physics, Harvard University, Cambridge, MA 02138; and
5. ^bHefei National Laboratory for Physical Sciences at the Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230026, China

1. Edited by Stefan W. Hell, Max Planck Institute for Biophys. Chemistry, Goettingen, Germany, and accepted by the Editorial Board July 7, 2012 (received for review February 1, 2012) 
   
   
   
   
    Imaging membranes in live cells with nanometer-scale resolution promises to reveal ultrastructural dynamics of organelles that are essential for cellular functions. In this work, we identified photoswitchable membrane probes and obtained super-resolution fluorescence images of cellular membranes. We demonstrated the photoswitching capabilities of eight commonly used membrane probes, each specific to the plasma membrane, mitochondria, the endoplasmic recticulum (ER) or lysosomes. These small-molecule probes readily label live cells with high probe densities. Using these probes, we achieved dynamic imaging of specific membrane structures in living cells with 30–60 nm spatial resolution at temporal resolutions down to 1–2 s. Moreover, by using spectrally distinguishable probes, we obtained two-color super-resolution images of mitochondria and the ER. We observed previously obscured details of morphological dynamics of mitochondrial fusion/fission and ER remodeling, as well as heterogeneous membrane diffusivity on neuronal processes.