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Multiphoton autofluorescence lifetime imaging of induced pluripotent stem cells

Aisada Uchugonova1

  • 1Saarland University, Department of Biophotonics and Laser Technology, Saarbrücken, GermanybJenLab GmbH, Jena, Germany.

Journal of Biomedical Optics
|July 1, 2017
PubMed

Insights

Multiphoton fluorescence lifetime imaging distinguished induced pluripotent stem cells (iPS) from feeder cells. This advanced imaging technique revealed unique autofluorescence signatures, aiding in stem cell characterization and differentiation studies.

Area of Science:

  • Biophotonics
  • Stem Cell Biology
  • Cellular Imaging

Background:

  • Induced pluripotent stem cells (iPS) hold great promise for regenerative medicine.
  • Distinguishing iPS cells from feeder cells is crucial for culture purity and downstream applications.
  • Autofluorescence (AF) imaging offers a label-free method for cellular analysis.

Purpose of the Study:

  • To investigate the utility of multiphoton fluorescence lifetime imaging (MP-FLIM) for characterizing iPS cell cultures.
  • To identify distinct AF lifetime signatures between iPS cells and supporting feeder cells.
  • To explore AF lifetime imaging for monitoring stem cell differentiation.

Main Methods:

  • Utilized a multiphoton fluorescence lifetime imaging tomograph (MPTflex) system.
  • Employed time-correlated single-photon counting for high temporal (250 ps) and spatial (submicron) resolution.
  • Analyzed autofluorescence from metabolic coenzymes NAD(P)H and flavins in iPS and feeder cells.

Main Results:

  • MP-FLIM successfully differentiated iPS cells from growth-arrested mouse embryonic fibroblast (MEF) feeder cells based on AF lifetime signatures.
  • Significant differences in AF lifetime were observed between iPS cells and feeder cells.
  • Distinct AF lifetime signatures were also identified between undifferentiated and differentiating stem cells.

Conclusions:

  • MP-FLIM is a powerful label-free technique for distinguishing iPS cells from feeder cells.
  • AF lifetime signatures provide valuable information for assessing stem cell identity and differentiation status.
  • This imaging approach has potential for quality control and research in pluripotent stem cell biology.

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