Live Imaging of Host-Pathogen Interactions in Zebrafish Embryos Infected with Mycobacterium abscessus

Samenvatting

Source: Bernut, A., et al. Deciphering and Imaging Pathogenesis and Cording of Mycobacterium abscessus in Zebrafish Embryos. J. Vis. Exp. (2015)

This video demonstrates a protocol that uses transgenic zebrafish embryos expressing fluorescent macrophages and infected with fluorescent Mycobacterium abscessus rough strain to visualize host-pathogen interactions. It reveals how extracellular cording enables immune evasion, promoting bacterial survival and host cell damage.

Protocol

1. Live imaging of M. abscessus infection

1. Mount tricaine-anesthetized embryos in 1% low melting point agarose in a 35 mm petri dish prior to epifluorescence microscopy observations. Use a glass bottom dish for inverted confocal microscopy or a single cavity depression slide for upright confocal microscopy. Orient the embryo to the desired position and cover the solidified agarose with fish water containing tricaine.
2. Use an epifluorescence microscope with a 10 X objective for sequential fluorescence acquisition and transmission images of the entire embryo. Alternatively, use a confocal microscope with 40X or 63X objectives to visualize the activity of immune cells after an infection.

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Low melting agarose	Sigma-Aldrich
Ethyl 3-aminobenzoate methanesulfonate salt (Tricaine)	Sigma-Aldrich	A5040