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Classification of Neural Stem Cell Activation State In Vitro using Autofluorescence
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Autofluorescence lifetime imaging classifies human B and NK cell activation state.

Rebecca L Schmitz1, Jeremiah M Riendeau1,2, Kelsey E Tweed1,2

  • 1Morgridge Institute for Research, Madison, WI, United States.

Frontiers in Bioengineering and Biotechnology
|April 21, 2025
PubMed
Summary

Optical metabolic imaging (OMI) offers a label-free method to assess immune cell function. This technique accurately distinguishes activated B cells and NK cells by analyzing their metabolic changes, aiding in cell therapy and immune profiling.

Keywords:
B cellsImmune activationNK cellsautofluorescenceimaging

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Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • Non-destructive, single-cell resolution tools are crucial for evaluating B cell and NK cell function.
  • Applications include adoptive cell therapy and immune profiling.
  • Optical metabolic imaging (OMI) is a label-free technique quantifying single-cell metabolism via autofluorescence of NAD(P)H and FAD.

Purpose of the Study:

  • To demonstrate OMI's capability in resolving metabolic differences between quiescent and activated primary human B and NK cells.
  • To assess the potential of OMI for label-free, non-destructive evaluation of immune cell activation status.

Main Methods:

  • Utilized Optical Metabolic Imaging (OMI) to measure autofluorescence intensity and lifetime of NAD(P)H and FAD in single cells.
  • Compared metabolic profiles of quiescent versus activated (IL-4/anti-CD40 for B cells; IL-12/IL-15/IL-18 for NK cells) primary human cells.
  • Employed random forest models to classify cell activation states based on OMI-derived metabolic parameters.

Main Results:

  • Stimulated B and NK cells exhibited altered metabolism, including increased free NAD(P)H, reduced redox state, and elevated lactate production.
  • A decrease in NAD(P)H mean fluorescence lifetime was observed in activated B and NK cells.
  • Random forest models achieved 93% accuracy in classifying B and NK cell activation states using OMI data.

Conclusions:

  • Autofluorescence lifetime imaging via OMI can accurately assess B and NK cell activation.
  • OMI provides a label-free and non-destructive method for immune cell functional analysis.
  • This approach holds promise for applications in adoptive cell therapy and immune profiling.