Colocalization of cellular nanostructure using confocal fluorescence and partial wave spectroscopy

John E. Chandler, Yolanda Stypula-Cyrus, Luay Almassalha, Greta Bauer, Leah Bowen, Hariharan Subramanian, Igal Szleifer, Vadim Backman*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

A new multimodal confocal microscope has been developed, which includes a parallel Partial Wave Spectroscopic (PWS) microscopy path. This combination of modalities allows molecular-specific sensing of nanoscale intracellular structure using fluorescent labels. Combining molecular specificity and sensitivity to nanoscale structure allows localization of nanostructural intracellular changes, which is critical for understanding the mechanisms of diseases such as cancer. To demonstrate the capabilities of this multimodal instrument, we imaged HeLa cells treated with valinomycin, a potassium ionophore that uncouples oxidative phosphorylation. Colocalization of fluorescence images of the nuclei (Hoechst 33342) and mitochondria (anti-mitochondria conjugated to Alexa Fluor 488) with PWS measurements allowed us to detect a significant decrease in nuclear nanoscale heterogeneity (Σ), while no significant change in Σ was observed at mitochondrial sites. In addition, application of the new multimodal imaging approach was demonstrated on human buccal samples prepared using a cancer screening protocol. These images demonstrate that nanoscale intracellular structure can be studied in healthy and diseased cells at molecular-specific sites. (Figure presented.).

Original languageEnglish (US)
Pages (from-to)377-384
Number of pages8
JournalJournal of Biophotonics
Volume10
Issue number3
DOIs
StatePublished - Mar 1 2017

Keywords

  • chromatin
  • confocal microscopy
  • hyperspectral microscopy
  • mitochondria
  • nanocytology

ASJC Scopus subject areas

  • Chemistry(all)
  • Materials Science(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Engineering(all)
  • Physics and Astronomy(all)

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