Non-destructive, label free identification of cell cycle phase in cancer cells by multispectral microscopy of autofluorescence

Jared M. Campbell*, Abbas Habibalahi, Saabah Mahbub, Martin Gosnell, Ayad G. Anwer, Sharon Paton, Stan Gronthos, Ewa Goldys

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

15 Citations (Scopus)
28 Downloads (Pure)


Background: Cell cycle analysis is important for cancer research. However, available methodologies have drawbacks including limited categorisation and reliance on fixation, staining or transformation. Multispectral analysis of endogenous cell autofluorescence has been shown to be sensitive to changes in cell status and could be applied to the discrimination of cell cycle without these steps.

Methods: Cells from the MIA-PaCa-2, PANC-1, and HeLa cell lines were plated on gridded dishes and imaged using a multispectral fluorescence microscope. They were then stained for proliferating cell nuclear antigen (PCNA) and DNA intensity as a reference standard for their cell cycle position (G1, S, G2, M). The multispectral data was split into training and testing datasets and models were generated to discriminate between G1, S, and G2 + M phase cells. A standard decision tree classification approach was taken, and a two-step system was generated for each line. 

Results: Across cancer cell lines accuracy ranged from 68.3% (MIA-PaCa-2) to 73.3% (HeLa) for distinguishing G1 from S and G2 + M, and 69.0% (MIA-PaCa-2) to 78.0% (PANC1) for distinguishing S from G2 + M. Unmixing the multispectral data showed that the autofluorophores NADH, FAD, and PPIX had significant differences between phases. Similarly, the redox ratio and the ratio of protein bound to free NADH were significantly affected.

Conclusions: These results demonstrate that multispectral microscopy could be used for the non-destructive, label free discrimination of cell cycle phase in cancer cells. They provide novel information on the mechanisms of cell-cycle progression and control, and have practical implications for oncology research.

Original languageEnglish
Article number1242
Pages (from-to)1-11
Number of pages11
JournalBMC Cancer
Issue number1
Publication statusPublished - 21 Dec 2019

Bibliographical note

Copyright The Author(s) 2019. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.


  • Cancer
  • Cell cycle
  • Cell phase
  • Hyperspectral
  • Multispectral
  • Neoplasia


Dive into the research topics of 'Non-destructive, label free identification of cell cycle phase in cancer cells by multispectral microscopy of autofluorescence'. Together they form a unique fingerprint.

Cite this