May. 23, 2017
News

High-Resolution Microscopy: A Precise Look into the Depths of Cells

  • Live yeast cells embedded in agarose. From left to right: conventional fluorescence, conventionally treated and csiLSFM. The bar has a width of 1 µm.  (Image: Goethe University / Stelzer group)Live yeast cells embedded in agarose. From left to right: conventional fluorescence, conventionally treated and csiLSFM. The bar has a width of 1 µm. (Image: Goethe University / Stelzer group)

Is it possible to observe how fish embryos develop into trout, carp or salmon at cell level? Researchers at the Goethe-University in Frankfurt Germany have managed to achieve this by combining a special form of fluorescence microscopy with a second microscopy technology. The new high resolution light microscope enables fascinating exploration of the interior of cells.  

But what’s new about that? Using traditional confocal fluorescence microscopy it is already possible to observe organisms very precisely and vividly through differentiation between the cells. Researchers mark the cell molecules with fluorescent markers which cause a light beam to shine. A camera records the three dimensional distribution of the fluorophores’ lit up molecules.          

Ernst Stelzer, professor at the Institute for Cell Biology and Neuroscience at the Goethe University and leader of research in the excellence cluster “Macromolecular complexes” at the same university, has made significant advances in fluorescence microscopy with his team. His light sheet fluorescence microscope (LSFM) is able to handle even sensitive samples such as fish embryos extremely carefully. An important step, as to date, traditional processes destroyed the pigments and the cells themselves in a very short time because of the energy in the photons.

In the technology developed by Stelzer micro-meter thin light sheets only focus on the area of the cell that the researcher wants to look at more closely. The camera takes pictures of the sample from different sides and angles. Computers put the individual pictures together to create three dimensional complete images and films. “As we examine the cell samples in the most natural growth conditions possible, we can achieve very precise results” says Stelzer. But not only static images of cells can be measured and compared directly. This is also possible for dynamic changes in the environment or for genetic mutations. 

Original publication:
Bo-Jui Chang, Victor Didier Perez Meza, and Ernst H. K. Stelzer: csiLSFM combines light-sheet fluorescence microscopy and coherent structured illumination for a lateral resolution below 100 nm, Proc Natl Acad Sci U S A, 114(19):4869-4874 (2017) doi: 10.1073/pnas.1609278114

More information:
http://www.muk.uni-frankfurt.de

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