Airyscan super-resolution microscopy of mitochondrial morphology and dynamics in living tumor cells

Vladimir L. Kolossov, Mayandi Sivaguru, Joseph Huff, Katherine Luby, Kaviamuthan Kanakaraju, H. Rex Gaskins

Research output: Contribution to journalArticlepeer-review

Abstract

Mitochondrial morphology is regulated by continuous fusion-and-fission events that are essential for maintaining normal function. Despite the prominence of mitochondrial function in energy generation and cell signaling, understanding of processes of fusion and fission dynamics has been hampered by the lack of high-resolution optical systems that accommodate live-cell imaging. We have examined different confocal modalities in terms of resolution and signal-to-noise ratio (SNR) in a point scanning confocal microscope with Airyscan super-resolution (AS-SR). Results indicated that Airyscan (AS) provided speed, super-resolution, and high SNR. This modality was then used for monitoring mitochondrial dynamics in live tumor cells modified to harbor green-fluorescent protein localized to mitochondria. We then compared regular AS and fast-Airyscan modalities in terms of gentleness on the live-cell samples. The fast mode provided unprecedented imaging speed that permits monitoring dynamics both in 2D and also in three-dimensional dataset with time lapses (4D). Alterations to the mitochondrial network in U87 glioblastoma cells occurred within seconds and the cells were not affected by modest inhibition of fission. The super-resolution permitted quantitative measurements of mitochondrial diameter with a precision that enabled detection of significant differences in mitochondrial morphology between cell lines. We have observed swelling of mitochondrial tubules in A549 lung cancer cells after 2 hr treatment with deoxynyboquinone, an ROS-generating pharmacologic drug. We also tested different 3D analytical parameters and how they can affect morphometric quantitation. The AS-SR imaging enabled high-speed imaging of mitochondrial dynamics without the compromise to cell morphology or viability that is common with conventional fluorescence imaging due to photo-oxidation.

Original languageEnglish (US)
Pages (from-to)115-128
Number of pages14
JournalMicroscopy research and technique
Volume81
Issue number2
DOIs
StatePublished - Feb 1 2018

Keywords

  • ROS
  • analysis of mitochondrial morphology
  • cancer cells
  • fission
  • live-cell imaging
  • super-resolution microscopy

ASJC Scopus subject areas

  • Anatomy
  • Histology
  • Instrumentation
  • Medical Laboratory Technology

Fingerprint Dive into the research topics of 'Airyscan super-resolution microscopy of mitochondrial morphology and dynamics in living tumor cells'. Together they form a unique fingerprint.

Cite this