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Related Experiment Videos

Quantitation of bacterial adherence by image analysis.

R Barthelson1, C Hopkins, A Mobasseri

  • 1Neose Technologies, Inc., Horsham, PA 19044, USA. rbarthelson@neose.com

Journal of Microbiological Methods
|October 16, 1999
PubMed
Summary

Quantifying bacterial adherence to host cells is difficult. This study presents a digital imaging method using immunocytochemistry to accurately count pathogenic bacteria, aiding in adherence studies.

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Area of Science:

  • Microbiology
  • Cell Biology
  • Biotechnology

Background:

  • Quantifying pathogenic bacterial adherence to host cells in vitro is crucial for understanding infection mechanisms but presents significant enumeration challenges.
  • Existing methods for bacterial counting can be labor-intensive and difficult to scale for high-throughput studies.

Purpose of the Study:

  • To develop and validate a novel digital imaging and computer-aided recognition method for accurate quantitation of bacteria attached to cultured eukaryotic cells.
  • To establish a reliable and efficient technique for studying bacterial-host cell interactions in vitro.

Main Methods:

  • Developed a protocol utilizing immunocytochemistry to selectively stain pathogenic bacteria while leaving host cells largely unstained.
  • Employed digital imaging and computer-aided recognition software for automated bacterial enumeration on host cell monolayers.

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  • Validated the method across five key bacterial species: Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus, and Chlamydia pneumoniae.

    • Main Results:

    • The developed method provides accurate and reproducible quantitation of adherent bacteria.
    • Demonstrated successful application by studying the adherence of Haemophilus influenzae and Streptococcus pneumoniae to respiratory tract epithelial cell lines.
    • The immunocytochemical staining effectively differentiated bacteria from host cells, facilitating precise counting.

    Conclusions:

    • Digital imaging and computer-aided recognition offer a robust solution for quantifying bacterial adherence in vitro.
    • This technique simplifies and enhances the efficiency of bacterial adherence assays, supporting research into pathogenic mechanisms.
    • The method is versatile and applicable to various pathogenic bacteria and host cell types relevant to infectious diseases.