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4D Super-Resolution Microscopy with Conventional Fluorophores and Single Wavelength Excitation in Optically Thick Cells and Tissues

Research output: Contribution to journalArticle

  • D Baddeley
  • D Crossman
  • S Rossberger
  • J Cheyne
  • J Montgomery
  • D Isuru
  • ID Jayasinghe
  • C Cremer
  • MB Cannell
  • C Soeller
Original languageEnglish
Article numbere20645
Pages (from-to)1-10
Number of pages10
JournalPLoS ONE
Volume6
DOIs
DatePublished - 2011

Abstract

BACKGROUND: Optical super-resolution imaging of fluorescently stained biological samples is rapidly becoming an important tool to investigate protein distribution at the molecular scale. It is therefore important to develop practical super-resolution methods that allow capturing the full three-dimensional nature of biological systems and also can visualize multiple protein species in the same sample. METHODOLOGY/PRINCIPAL FINDINGS: We show that the use of a combination of conventional near-infrared dyes, such as Alexa 647, Alexa 680 and Alexa 750, all excited with a 671 nm diode laser, enables 3D multi-colour super-resolution imaging of complex biological samples. Optically thick samples, including human tissue sections, cardiac rat myocytes and densely grown neuronal cultures were imaged with lateral resolutions of ∼15 nm (std. dev.) while reducing marker cross-talk to

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