Super-Resolved Fluorescence Microscopy

For this breakthrough with important applications to biology, Eric Betzig won the 2014 Nobel Prize in Chemistry.

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Super-Resolved Fluorescence Microscopy reveals high-resolution 3D images of subcellular activity in real time with minimal damage to cells, allowing biologists to study the activity of molecules, cells and embryos in fine detail over longer periods than ever before. 

Breaking the diffraction barrier

Eric Betzig earned a BS in Physics at California Institute of Technology (1983). He obtained an MS (1985) and PhD (1988) in Applied and Engineering Physics from Cornell University. His thesis focused on the development of the first super-resolution optical microscope, called the near-field scanning optical microscope (NSOM) [3]. He then joined Bell Labs in Murray Hill and worked to improve the NSOM technology he had helped develop at Cornell.

Betzig worked with Jay Trautman, Tim Harris and others to refine the NSOM technology and try out new applications. Together, they developed a scanning microscope featuring a new near-field probe capable of generating images of samples at a resolution of 12 nanometers — better than the diffraction limit. The probe also yielded signals more than 100 times larger than those reported previously. The first to break the diffraction barrier, the powerful microscope was seen as a new means to inspect integrated circuits and examine living cells.

The microscope was recognized by R&D Magazine as one of the 100 most significant inventions of 1991.

NSOM technology applications

Collaborations with Trautman, Harris, Harald Hess and Rob Chichester allowed Betzig to explore several applications for the NSOM technology, including high-density data storage, semiconductor spectroscopy and super-resolution fluorescence cell imaging[4]. This exploration yielded key breakthroughs that brought Betzig closer to his dream of imaging molecules within living cells.

In 1992, Betzig and Trautman used their microscope as the basis for developing a new high-density magneto-optic storage technique. The technique supported data densities of 45 billion bits per square inch — nearly 100 times the density provided by the best commercial magneto-optic methods of the day, and 300 times the density supported by contemporary magnetic-storage capabilities. 


“Because of Bell Labs' history and the brilliance of everyone around me, I felt like I was on probation from the time I got there. Two years in, I wrote in my self-evaluation that if I didn’t have a breakthrough in the next year, they wouldn’t need to fire me because I would quit.” — Eric Betzig

Next step: fluorescence

In November 1993, Betzig and Chichester reported the first single molecule microscopy at room temperature, the first localization of single molecules (to ~12 nm), and the first determination of single molecule dipole orientations.  

The seeds of PALM microscopy

In 1994, Betzig and Harald Hess brought the near-field technology together with Hess’s low temperature scan probe technology. The combination of technologies allowed them to see luminescent centers of molecules [5] with high spatial resolution.

In the late 1990s, Betzig left Bell Labs to pursue other interests, and went to work at his father’s machine tool company for several years. He developed and marketed a rapid robotic machine tool that was technically successful but did not find enough customers to be profitable. But during that interval, other researchers had made progress in fluorescent protein chemistry, and these advances provided a new way to realize super-resolution microscopy without requiring a near-field probe. 

Return to research

Betzig and Hess began collaborating again on their ideas, and built a super-resolution microscope that formed the basis for the first paper on photo-activated localization microscopy (PALM) technology in 2006. The PALM technology allowed Betzig and Hess to optically image intracellular proteins at nanometer spatial resolution [6].  Betzig and Hess continue to collaborate at the Janelia Farms campus of the Howard Hughes Medical Institute (HHMI).

Betzig, Stefan W Hell and William E Moerner were awarded the the 2014 Nobel Prize in Chemistry for their development of Super-Resolved Fluorescence Microscopy [1]. 


[1] “Lattice Light Sheet is a Leap Forward for Microscopy” Web. 12 Nov 2014.

[2] "Eric Betzig” Janelia Farm Research campus. Web. 5 Nov 2014.

[3] "Eric Betzig” Janelia Farm Research campus. Web. 5 Nov 2014.

[4] "Eric Betzig and Harald Hess: Developing Palm Microscopy" Web. 7 Nov 2014.

[5] E Betzig, GH Patterson, R Sougrat, OW Lindwasser, S Olenych, JS Bonifacino, MW Davidson, J Lippincott-Schwartz, HF Hess, "Imaging intracellular fluorescent proteins at nanometer resolution", Science 313, 1642 (2006).

[6] “Eric Betzig - Facts". Nobel Media AB 2014. Web. 5 Nov 2014.