Photon Yield Enhancement of Red Fluorophores at Cryogenic Temperatures. Issue 14 (15th May 2018)

• Record Type: Journal Article
• Authors: Hulleman, Christiaan N.; Li, Weixing; Gregor, Ingo; Rieger, Bernd; Enderlein, Jörg
• Is Part Of: Chemphyschem. Volume 19:Issue 14(2018)
• Page Start: 1774
• Page End: 1780
• Publication Date: 2018-05-15
• Subjects: single-molecule photon yield -- cryo-fluorescence microscopy -- single-molecule localization precision -- super-resolution microscopy; Chemistry, Physical and theoretical -- Periodicals; 541.05
• DOI: 10.1002/cphc.201800131
• Languages: English
• ISSNs: 1439-4235

Abstract

Abstract: Single Molecule Localization Microscopy has become one of the most successful and widely applied methods of Super‐resolution Fluorescence Microscopy. Its achievable resolution strongly depends on the number of detectable photons from a single molecule until photobleaching. By cooling a sample from room temperature down to liquid nitrogen temperatures, the photostability of dyes can be enhanced by more than 100 fold, which results in an improvement in localization precision greater than 10 times. Here, we investigate a variety of fluorescent dyes in the red spectral region, and we find an average photon yield between 3.5 ⋅ 10 6 to 11 ⋅ 10 6 photons before bleaching at liquid nitrogen temperatures, corresponding to a theoretical localization precision around 0.1 nm. Abstract : In single‐molecule localization microscopy the resolution depends strongly on the number of detected photons from a single molecule until photobleaching. At cryogenic temperatures, photochemical reaction rates are reduced, and therefore, the authors investigate the photostability of six red fluorescent dyes at about 89 K. At cryogenic temperatures, the photon yield is increased by a factor of 100, resulting in a tenfold enhancement of the localization precision. The six different dyes have an average photon yield ranging from 3.5 ⋅ 10 6 to 11 ⋅ 10 6 photons.