A method for the enumeration of bacterial adhesion to epithelial cells using image analysis.
Computerised image analysis was utilised to enumerate the attachment of Staphylococcus epidermidis to HEp2 cell monolayers. A differential staining technique was employed such that individual staphylococcal cells stood out in sharp contrast against the uneven cell surface and granular contents of the epithelial cells. The primary image analysis operation involved subtracting an out-of-focus image from an in-focus image of the bacteria on the monolayer, thereby accentuating the bacterial image. Enumeration, using a particle counting routine, was rapid and reproducible, facilitating counting in excess of 700 bacteria per field at x 500 magnification. The computerised programme compared favourably with manual counting and would provide a rapid, objective and morphologically discriminatory method for evaluating bacterial attachment to various tissues.
Evans-Hurrell
J A
JA
Department of Pharmaceutical Sciences, University of Nottingham, UK.
Adler
J
J
Denyer
S
S
Rogers
T G
TG
Williams
P
P
D001422:Bacterial Adhesion; D001431:Bacteriological Techniques; D004847:Epithelial Cells; D004848:Epithelium / Q000382:microbiology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D013194:Staining and Labeling; D013212:Staphylococcus epidermidis / Q000502:physiology*; D014407:Tumor Cells, Cultured