Journal
NANO LETTERS
Volume 14, Issue 7, Pages 4171-4175Publisher
AMER CHEMICAL SOC
DOI: 10.1021/nl501870p
Keywords
Fluorescence cryo-microscopy; vitrification; localization microscopy; single molecule blinking; low temperature; frozen-hydrated cells
Categories
Funding
- Wellcome Trust [090895/Z/09/Z, 096144/Z/11/Z, 090532/Z/09/Z, 091911]
- EMBO Fellowship [ALTF 642-2011]
- Cancer Research UK programme grant [A10976]
- Wellcome Trust [096144/Z/11/Z, 090895/Z/09/Z] Funding Source: Wellcome Trust
- Cancer Research UK [10976, 17721] Funding Source: researchfish
- MRC [MR/K01577X/1] Funding Source: UKRI
Ask authors/readers for more resources
We introduce a super-resolution technique for fluorescence cryo-microscopy based on photoswitching of standard genetically encoded fluorescent marker proteins in intact mammalian cells at low temperature (81 K). Given the limit imposed by the lack of cryo-immersion objectives, current applications of fluorescence cryo-microscopy to biological specimens achieve resolutions between 400-500 nm only. We demonstrate that the single molecule characteristics of reversible photobleaching of mEGFP and mVenus at liquid nitrogen temperature are suitable for the basic concept of single molecule localization microscopy. This enabled us to perform super-resolution imaging of vitrified biological samples and to visualize structures in unperturbed fast frozen cells for the first time with a structural resolution of similar to 125 nm (average single molecule localization accuracy similar to 40 nm), corresponding to a 3-5 fold resolution improvement.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available