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Label-Free Quantification of Apoptosis and Necrosis Using Stimulated Raman Scattering Microscopy.

Shivam Mahapatra1, Shreya B Shivpuje2, Helen C Campbell1

  • 1James Tarpo Jr. and Margaret Tarpo Department of Chemistry, Purdue University, 560 Oval Drive, West Lafayette, Indiana 47907, United States.

Analytical Chemistry
|June 13, 2025
PubMed
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This summary is machine-generated.

Stimulated Raman scattering (SRS) microscopy rapidly detects cell death in biomanufacturing. This label-free method monitors apoptosis and necrosis in Chinese hamster ovary (CHO) cells, improving production quality.

Area of Science:

  • Biotechnology
  • Cell Biology
  • Microscopy

Background:

  • Chinese hamster ovary (CHO) cells are vital for recombinant protein production.
  • Cell death, especially apoptosis, during manufacturing reduces yield and quality.
  • Rapid, label-free methods are needed to monitor cell health in bioreactors.

Purpose of the Study:

  • To evaluate Stimulated Raman scattering (SRS) microscopy for label-free monitoring of cell death.
  • To differentiate between apoptotic and necrotic transitions in live cells.
  • To develop a quantitative analysis pipeline for cell death signatures.

Main Methods:

  • Utilized SRS microscopy for label-free analysis of cellular composition.
  • Developed a single-cell analysis pipeline to extract chemotypic and phenotypic signatures.

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  • Applied the method to Chinese hamster ovary (CHO) cells under various stressors.

    • Main Results:

    • SRS microscopy accurately distinguished between apoptotic and necrotic cells.
    • Apoptotic cells showed higher protein concentrations, necrotic cells showed lower.
    • The analysis pipeline identified distinct cell subpopulations and was generalized to other cell types.

    Conclusions:

    • SRS microscopy is a robust, noninvasive tool for rapid cell death monitoring.
    • This method enhances quality control in biopharmaceutical production.
    • Findings are applicable to diverse biological contexts and cell death evaluations.