Fluorescence Polarization Control for On-Off Switching of Single Molecules at Cryogenic Temperatures
RNA Therapeutics Institute; Graduate School of Biomedical Sciences, Interdisciplinary Graduate Program
Bioimaging and Biomedical Optics | Biophysics | Molecular Biology | Structural Biology
Light microscopy, allowing sub-diffraction-limited resolution, has been among the fastest developing techniques at the interface of biology, chemistry, and physics. Intriguingly no theoretical limit exists on how far the underlying measurement uncertainty can be lowered. In particular data fusion of large amounts of images can reduce the measurement error to match the resolution of structural methods like cryo-electron microscopy. Fluorescence, although reliant on a reporter molecule and therefore not the first choice to obtain ultraresolution structures, brings highly specific labeling of molecules in a large assembly to the table and inherently allows the detection of multiple colors, which enables the interrogation of multiple molecular species at the same time in the same sample. Here, the problems to be solved in the coming years, with the aim of higher resolution, are discussed, and what polarization depletion of fluorescence at cryogenic temperatures can contribute for fluorescence imaging of biological samples, like whole cells, is described.
STED, cryogenic, polarization, single molecules, super-resolution
DOI of Published Version
Small Methods. 2018 Sep 11;2(9). doi: 10.1002/smtd.201700323. Epub 2018 Apr 30. Link to article on publisher's site
Hulleman, Christiaan N.; Huisman, Maximiliaan; Moerland, Robert J.; Grünwald, David; Stallinga, Sjoerd; and Rieger, Bernd, "Fluorescence Polarization Control for On-Off Switching of Single Molecules at Cryogenic Temperatures" (2018). RNA Therapeutics Institute Publications. 60.