JoVE Encyclopedia of Experiments
Immunology
0 views • 3:48 min • July 8th, 2025
Take a microtiter plate, and add serial dilutions of novel antimicrobial peptides or AMPs — the test compounds.
Designate a subset of wells as growth controls, and add sterile water without the AMPs.
Add a pathogenic fungus suspended in growth media into selected wells.
Add only the media, devoid of the fungus, in a subset of wells designated as blank controls.
Seal the plate and incubate under humid conditions conducive to fungal growth.
Post-incubation, observe the wells under a microscope.
The growth controls appear turbid due to fungal growth in the absence of AMPs. Turbidity across all dilutions of an AMP indicates its ineffectiveness against the fungus.
The blank controls, devoid of fungal growth, appear clear. Compare test wells with the blank controls to confirm the absence of growth, indicating the antifungal activity of AMPs.
The lowest concentration of an AMP that inhibits fungal growth is termed the minimum inhibitory concentration, or MIC.
The antifungal assay is performed in triplicate in 96-well polystyrene microplates using a 2-fold serial dilution of each peptide and control antifungal and a final volume of 50 microliters per well.
The most crucial step in the prot
View the full transcript and gain access to thousands of scientific videos