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A procedure for dissociating Ayre scrape samples

R C Leif, S Nordqvist, S Clay

    The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
    |July 1, 1977
    PubMed
    Summary
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    This study optimized cervical cell dissociation for improved analysis. Enhanced enzymatic and chemical treatments yield significantly higher percentages of single cervical cells, enabling automated scanning systems.

    Area of Science:

    • Cytopathology
    • Biochemistry
    • Cell Biology

    Background:

    • Accurate cervical cell analysis is crucial for disease detection.
    • Current methods face challenges with cell clumping and dissociation.
    • Improving cell suspension quality is key for automated analysis.

    Purpose of the Study:

    • To develop an optimized protocol for dissociating cervical cell suspensions.
    • To enhance the percentage of single cervical cells for improved analysis.
    • To assess the feasibility of automated scanning systems with improved cell preparations.

    Main Methods:

    • Utilized Centrifugal Cytology rotor for glutaraldehyde-fixed dispersions.
    • Employed a combination of enzymatic (ribonuclease, Varidase) and chemical (dithiothreitol, iodoacetic acid, EDTA) treatments.

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  • Developed an RPG II program for recording and analyzing dissociation results.
  • Assessed cell dissociation and recovery rates for white blood cells and cervical cells.

    • Main Results:

    • Optimized protocol achieved monodisperse white blood cells.
    • Achieved 81% single cervical cells, increasing to 98% with nuclear separation criteria.
    • Control preparations showed 92% single white blood cells and 63% single cervical cells.
    • The improved cell dispersion is suitable for automated scanning systems.

    Conclusions:

    • The optimized dissociation protocol significantly improves cervical cell preparation quality.
    • Higher percentages of single cells enhance the reliability of cytopathological analysis.
    • The enhanced cell suspensions are compatible with automated slide scanning technologies.