Encyclopedia of Experiments: Immunology
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Transcript
To quantify Helicobacter pylori colonization in an infected mouse stomach, begin with the flattened stomach tissue containing the body and antrum regions, suspended in nutrient media to support bacterial viability.
Homogenize to break down the gastric tissue with the mucus layer, releasing Helicobacter pylori from the tissue into media. Perform serial dilutions of the resulting gastric homogenate using media.
Obtain dried horse blood agar, HBA, plates, divided into multiple segments. The HBA plates are supplemented with a mixture of antibiotics. Transfer each homogenate dilution onto separate segments of the plate and spread it evenly.
Post-drying, position the plates in an inverted position within a humidified anaerobic culture jar with specialized gas mixtures. Incubate.
The microaerophilic environment provided by the anaerobic culture jar in conjunction with the HBA nutrients facilitates Helicobacter pylori to grow and form colonies. The antibiotics in HBA inhibit the growth of bacterial species from the endogenous gastric microbiota.
Count Helicobacter pylori colonies on HBA plate segments falling within a specific countable range.
Calculate the Helicobacter pylori load in the gastric tissue, indicating the extent of bacterial colonization within the tissue.
