Super-resolution fluorescence imaging of organelles in live cells with photoswitchable membrane probes.
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.
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
Sang-Hee
SH
Shim
Chenglong
C
Xia
Guisheng
G
Zhong
Hazen P
HP
Babcock
Joshua C
JC
Vaughan
Bo
B
Huang
Xun
X
Wang
Cheng
C
Xu
Guo-Qiang
GQ
Bi
Xiaowei
X
Zhuang
D001896:Boron Compounds / Q000737:chemistry; D002232:Carbocyanines / Q000737:chemistry; D002462:Cell Membrane / Q000648:ultrastructure; D003712:Dendrites / Q000648:ultrastructure; D004721:Endoplasmic Reticulum / Q000648:ultrastructure; D005456:Fluorescent Dyes / Q000737:chemistry*; D006624:Hippocampus / Q000166:cytology; D008051:Lipid Bilayers; D008247:Lysosomes / Q000648:ultrastructure; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D008928:Mitochondria / Q000648:ultrastructure; D049329:Nanostructures / Q000648:ultrastructure*; D009474:Neurons / Q000648:ultrastructure; D015388:Organelles / Q000648:ultrastructure*; D011554:Pseudopodia / Q000648:ultrastructure; D013269:Stochastic Processes
C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D001896:Boron Compounds; D002232:Carbocyanines; D005456:Fluorescent Dyes; D008051:Lipid Bilayers