Establishing super-resolution imaging for proteins in diatom biosilica

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

Abstract

The intricate, genetically controlled biosilica nano- and micropatterns produced by diatoms are a testimony for biology's ability to control mineral formation (biomineralization) at the nanoscale and regarded as paradigm for nanotechnology. Previously, several protein families involved in diatom biosilica formation have been identified, and many of them remain tightly associated with the final biosilica structure. Determining the locations of biosilica-associated proteins with high precision is, therefore expected to provide clues to their roles in biosilica morphogenesis. To achieve this, we introduce here single-molecule localization microscopy to diatoms based on photo-activated light microscopy (PALM) to overcome the diffraction limit. We identified six photo-convertible fluorescent proteins (FPs) that can be utilized for PALM in the cytoplasm of model diatom Thalassiosira pseudonana. However, only three FPs were also functional when embedded in diatom biosilica. These were employed for PALM-based localization of the diatom biosilica-associated protein Silaffin-3 (tpSil3) with a mean precision of 25 nm. This allowed for the identification of distinct accumulation areas of Sil3 in the biosilica, which cannot be resolved by confocal fluorescence microscopy. The enhanced microscopy technique introduced here for diatoms will aid in elucidating the molecular mechanism of silica biomineralization as well as other aspects of diatom cell biology.

Details

Original languageEnglish
Article number36824
Number of pages8
JournalScientific reports
Volume6
Publication statusPublished - 9 Nov 2016
Peer-reviewedYes

External IDs

Scopus 84995489550
ORCID /0000-0002-6209-2364/work/142237662
ORCID /0000-0002-4533-8860/work/142241022

Keywords

Keywords

  • SINGLE-MOLECULE LOCALIZATION, THALASSIOSIRA-PSEUDONANA, FLUORESCENT PROTEINS, SILICA MORPHOGENESIS, ELECTRON-MICROSCOPY, LIGHT, RESOLUTION, TRANSFORMATIONS, SILAFFINS, STORM