Inkjet-printed micro-calibration standards for ultraquantitative Raman spectral cytometry

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Cell Biology

Background:

• Accurate quantitation of intracellular biomolecules is crucial for understanding cellular function and disease.
• Existing single-cell analysis methods often lack absolute quantitation capabilities.

Purpose of the Study:

• To develop and validate a Raman spectroscopy-based cytometric platform for absolute mass quantitation of endogenous biomolecules in single cells.
• To establish ultraquantitative calibration using engineered micro-calibration standards.

Main Methods:

• Development of a Raman-based cytometric analysis platform.
• Engineering of single-cell-sized micro-calibration standards with known biomolecular composition via inkjet printing.
• Characterization of human skin fibroblasts and porcine alveolar macrophages.

Main Results:

• First report of Raman-based quantitation of absolute mass of multiple endogenous biomolecules within single cells.
• Demonstrated clinical feasibility by analyzing cell populations in lipid storage disorders (Niemann-Pick disease, drug-induced phospholipidosis).
• Successful calibration using precisely fabricated micro-calibration standards.

Conclusions:

• The developed microanalytical platform provides absolute quantitation of single-cell biomolecular content.
• This technology has significant potential for diagnostic assays, pathological analysis, and pharmaco/toxicokinetic studies.
• Represents a foundational advancement in single-cell analysis and biomedical research.