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An Expandable Mechanopharmaceutical Device (3): a Versatile Raman Spectral Cytometry Approach to Study the Drug Cargo Capacity of Individual Macrophages

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Abstract

Purpose

To improve cytometric phenotyping abilities and better understand cell populations with high interindividual variability, a novel Raman-based microanalysis was developed to characterize macrophages on the basis of chemical composition, specifically to measure and characterize intracellular drug distribution and phase separation in relation to endogenous cellular biomolecules.

Methods

The microanalysis was developed for the commercially-available WiTec alpha300R confocal Raman microscope. Alveolar macrophages were isolated and incubated in the presence of pharmaceutical compounds nilotinib, chloroquine, or etravirine. A Raman data processing algorithm was specifically developed to acquire the Raman signals emitted from single-cells and calculate the signal contributions from each of the major molecular components present in cell samples.

Results

Our methodology enabled analysis of the most abundant biochemicals present in typical eukaryotic cells and clearly identified “foamy” lipid-laden macrophages throughout cell populations, indicating feasibility for cellular lipid content analysis in the context of different diseases. Single-cell imaging revealed differences in intracellular distribution behavior for each drug; nilotinib underwent phase separation and self-aggregation while chloroquine and etravirine accumulated primarily via lipid partitioning.

Conclusions

This methodology establishes a versatile cytometric analysis of drug cargo loading in macrophages requiring small numbers of cells with foreseeable applications in toxicology, disease pathology, and drug discovery.

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Author information

Authors and Affiliations

  1. Department of Pharmaceutical Sciences, University of Michigan College of Pharmacy, Ann Arbor, Michigan, 48109, USA

    Vernon LaLone & Gus R. Rosania

  2. Consulting for Statistics, Computing, and Analytics Research (CSCAR) Center, University of Michigan, Ann Arbor, Michigan, 48109, USA

    Márcio A. Mourão & Kerby Shedden

  3. Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, Michigan, 48109, USA

    Theodore J. Standiford & Kathleen A. Stringer

  4. Department of Surgery, University of Michigan Medical Center, Ann Arbor, Michigan, 48109, USA

    Krishnan Raghavendran

  5. Department of Clinical Pharmacy, University of Michigan College of Pharmacy, Ann Arbor, Michigan, 48109, USA

    Kathleen A. Stringer

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  1. Vernon LaLone
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  2. Márcio A. Mourão
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  3. Theodore J. Standiford
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  7. Gus R. Rosania
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Correspondence to Gus R. Rosania.

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LaLone, V., Mourão, M.A., Standiford, T.J. et al. An Expandable Mechanopharmaceutical Device (3): a Versatile Raman Spectral Cytometry Approach to Study the Drug Cargo Capacity of Individual Macrophages. Pharm Res 36, 2 (2019). https://doi.org/10.1007/s11095-018-2540-0

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  • DOI: https://doi.org/10.1007/s11095-018-2540-0

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