Three-dimensional super-resolution fluorescence imaging of DNA

Recent advances in fluorescence super-resolution microscopy are providing important insights into details of cellular structures. To acquire three dimensional (3D) super-resolution images of DNA, we combined binding activated localization microscopy (BALM) using fluorescent double-stranded DNA inter...

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Bibliographic Details
Published inbioRxiv
Main Authors Yardimci, Sevim, Burnham, Daniel R, Terry, Samantha Y A, Yardimci, Hasan
Format Paper
LanguageEnglish
Published Cold Spring Harbor Cold Spring Harbor Laboratory Press 07.10.2019
Subjects
Chromosomes
Deoxyribonucleic acid
DNA
Localization
Metaphase
Microscopy
Microspheres
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Summary:Recent advances in fluorescence super-resolution microscopy are providing important insights into details of cellular structures. To acquire three dimensional (3D) super-resolution images of DNA, we combined binding activated localization microscopy (BALM) using fluorescent double-stranded DNA intercalators and optical astigmatism. We quantitatively establish the advantage of mono- over bis-intercalators before demonstrating the approach by visualizing single DNA molecules stretched between microspheres at various heights. Finally, the approach is applied to the more complex environment of intact and damaged metaphase chromosomes, unravelling their structural features.
DOI:10.1101/796151