Prof. João Sérgio Seixas de Melo Profile

Prof. João Sérgio Seixas de Melo

at Univ. de Coimbra

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Proceedings Article | 12 March 2024 Presentation + Paper

Metabolic imaging with FLIM/PLIM indirubin derivatives: an ultrafast window into cellular metabolism and oxygen consumption

D. N. S. dos Santos, N. Naskar, K. Reess, E. Delgado-Pinar, J. Seixas de Melo, A. Rueck

Proceedings Volume 12847, 128470H (2024) https://doi.org/10.1117/12.3002527

KEYWORDS: Oxygen, Fluorescence lifetime imaging, Biological imaging, Ultrafast imaging, Microscopy

Read Abstract +

Metabolic Fluorescence Lifetime Imaging Microscopy (FLIM) algorithms have emerged as powerful tools for unraveling the intricate dynamics of cellular and tissue metabolism. FLIM algorithms offer unique insights into cellular metabolic processes that transcend traditional imaging methods. By combining Metabolic FLIM with Phosphorescence Lifetime Imaging Microscopy (PLIM), it becomes possible to assess cellular metabolic states, such as oxygen consumption, redox states, pH levels, and energy production pathways.

In our investigation, we employed a combination of 2-photon (2P) excited FLIM and PLIM techniques, along with timecorrelated single-photon counting (TCSPC) detection. Through this, we made a significant discovery of bromine indirubin derivatives that exhibit a PLIM/dFLIM signal in two living cell lines. Notably, indirubin is a natural dye, and though renowned for its anti-tumor properties, its mechanism of action remains to be fully investigated.

Under normoxic conditions, the PLIM signal of indirubin exhibited a value of 62 ns living cells, while under hypoxic conditions, it increased significantly to 107ns. This observation demonstrates the potential of these indirubins as highly reliable oxygen consumption sensors. Moreover, our investigation revealed that bromine indirubin had a profound impact on cellular metabolism, prompting a shift from oxidative phosphorylation to glycolysis.

Through our research, we aim to demonstrate that these techniques offer valuable insights into cellular metabolism, covering the way for deeper understanding and potential breakthroughs in various fields of biology and medicine.

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