JoVE Encyclopedia of Experiments
Microbiology
0 views • 4:46 min • January 30th, 2026
Take a mouse gingival section infected with pathogenic bacteria, pretreated to access bacterial DNA.
Apply a buffer with a digoxigenin-labeled DNA probe targeting bacterial DNA.
Place a coverslip and seal to prevent sample dehydration.
Introduce heat to denature the DNA, then incubate under hot and humid conditions to allow probe hybridization.
Chill the slide and remove the coverslip, and wash multiple times with buffer to remove unbound probes.
Add a blocking solution to prevent non-specific antibody binding.
Incubate with enzyme-conjugated antibodies that bind to the digoxigenin.
Wash to remove unbound antibodies.
Add an inhibitor to selectively inactivate the endogenous enzyme.
Apply a substrate that reacts with the enzyme to give a colored precipitate.
Rinse with water to remove excess substrate.
Add a dye to stain the nuclei.
Wash to remove excess dye.
Dehydrate in ethanol and treat with xylene to increase tissue transparency.
Mount the section.
Under a microscope, visualize the bacteria within the section.
Start the in situ by first immersing the slides in 2x SSC for 20 minutes. Meanwhile, dilute the labeled probe to 1 nanogram per microliter in hybridization buffer. For a negative control,
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