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Updated: Jan 14, 2026
Live Cell Imaging to Assess the Dynamics of Metaphase Timing and Cell Fate Following Mitotic Spindle Perturbations
Published on: September 20, 2019
Visualizing the chronicle of multiple cell fates using a near-IR dual-RNA/DNA-targeting probe
Linawati Sutrisno1, Gary J Richards2, Jack D Evans3
1International Center for Young Scientists (ICYS), National Institute for Materials Science, 1-1 Namiki, Tsukuba, Ibaraki 305-0044, Japan.
Abstract:
Early detection and late-stage cell fate assessment are key factors to develop therapeutic strategies, although current methods cannot capture early responses or distinguish multiple injury states, especially in ultraviolet-visible (UV-vis)-sensitive cells. Here, we introduce a method to simultaneously detect variations in RNA and DNA under near-infrared photoexcitation. Using a pyrazinacene-based probe (TEG8-N14), we unexpectedly achieved discrimination of multiple cell states, including apoptosis, necrosis, necroptosis, and senescence, based on RNA/DNA changes. Specifically, TEG8-N14 selectively stains necrotic cells in live samples, while after fixation, it allows detection of ultraearly senescence in UV-vis-sensitive cells, providing approximately twofold greater informational content than existing RNA or DNA fluorophores. These findings break current imaging barriers by enabling comprehensive visualization of single-cell fate histories without being affected by UV-vis or genetic manipulation.
Insights
This study introduces a new imaging method using a pyrazinacene probe (TEG8-N14) for simultaneous RNA and DNA detection. It effectively distinguishes multiple cell death states, including early senescence, in UV-vis-sensitive cells.
Area of Science:
- Cell Biology
- Biochemistry
- Molecular Imaging
Background:
- Current cell fate assessment methods struggle with early detection and distinguishing multiple injury states, particularly in UV-vis-sensitive cells.
- Limitations exist in capturing rapid cellular responses and differentiating diverse cell death pathways like apoptosis, necrosis, necroptosis, and senescence.
Purpose of the Study:
- To develop a novel imaging technique for simultaneous RNA and DNA detection under near-infrared photoexcitation.
- To enable discrimination of multiple cell states and capture early responses in UV-vis-sensitive cells.
Main Methods:
- Utilized a pyrazinacene-based probe (TEG8-N14) for near-infrared photoexcitation.
- Developed a method for simultaneous detection of RNA and DNA variations within single cells.
- Applied the probe to live and fixed UV-vis-sensitive cells to assess different cell fate outcomes.
Main Results:
- The TEG8-N14 probe successfully discriminated between apoptosis, necrosis, necroptosis, and senescence based on RNA/DNA changes.
- TEG8-N14 selectively stained necrotic cells in live samples.
- Post-fixation, the probe detected ultraearly senescence in UV-vis-sensitive cells, offering twofold greater information than existing fluorophores.
Conclusions:
- The developed method overcomes current imaging barriers for UV-vis-sensitive cells.
- This technique allows comprehensive visualization of single-cell fate histories without interference from UV-vis sensitivity or genetic manipulation.
- The probe provides enhanced informational content for detailed cell state analysis.
