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A microsample collection device for electrostatically sorted cells or particles and its preparative use for

W G Hinson, J L Pipkin, J L Hudson

    Cytometry
    |May 1, 1982
    PubMed
    Summary

    A novel collection device simplifies nuclear protein analysis by enabling precise sorting of cell nuclei based on DNA content. This method enhances sensitivity and reduces sample loss for detailed biochemical studies.

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    Area of Science:

    • Biochemistry
    • Cell Biology
    • Analytical Chemistry

    Background:

    • Accurate biochemical analysis of cellular components requires pure, well-preserved samples.
    • Flow cytometry is a powerful tool for cell sorting, but efficient collection of sorted nuclei can be challenging.
    • Previous methods for collecting sorted nuclei often led to sample degradation or loss, limiting downstream analysis.

    Purpose of the Study:

    • To develop and evaluate a specialized collection device for recovering sorted mammalian cell nuclei.
    • To simplify the biochemical analysis of proteins from nuclei sorted by DNA content.
    • To improve sample integrity and yield for sensitive proteomic studies.

    Main Methods:

    • Design and fabrication of a novel collection device for a sorting flow cytometer.

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  • Sorting of rat liver cell nuclei into distinct populations based on DNA content (G0+G1, S, G2+M).
  • Extraction of nuclear proteins and subsequent analysis using polyacrylamide gel isoelectric focusing and capillary isotachophoresis.

    • Main Results:

    • The collection device successfully recovered sorted nuclei with minimal degradation and loss.
    • Sufficient quantities of nuclei were collected to achieve adequate sensitivity and resolution for protein analysis.
    • Distinct nuclear protein profiles were obtained for different cell cycle phases (G0+G1, S, G2+M).

    Conclusions:

    • The developed collection device significantly simplifies the process of preparing sorted nuclei for biochemical analysis.
    • This method allows for sensitive and high-resolution protein analysis from specific nuclear populations.
    • The approach is valuable for studying cell cycle-dependent changes in nuclear protein composition.